- Volume 63, Issue Pt_2, 2013

The hyperthermophilic crenarchaeon Thermoproteus neutrophilus V24StaT was originally classified before sequence-based phylogenetic analysis became standard for bacterial taxonomy. Subsequent phylogenetic analyses by various groups have shown that strain V24StaT groups more closely with strains of the genus Pyrobaculum than with those in the genus Thermoproteus . Based on phylogenetic comparison of rRNA gene sequences and ribosomal proteins, we propose that strain V24StaT be reclassified as Pyrobaculum neutrophilum comb. nov., with the type strain V24StaT ( = DSM 2338T = JCM 9278T). An emended description of the genus Pyrobaculum is also presented.

A thorough phenotypic and genotypic analysis of 150 strains belonging to the Mycobacterium terrae complex resulted in the identification of a number of previously unreported sequevars (sqvs) within the species known to belong to the complex. For the species Mycobacterium arupense , three sqvs were detected in the 16S rRNA gene, six sqvs in the hsp65 gene and 15 sqvs in the rpoB gene; in Mycobacterium senuense two sqvs were present in each of the three genetic regions; in Mycobacterium kumamotonense four, two and nine sqvs were found, respectively, and in M. terrae three, four and six sqvs were found, respectively. The inappropriate inclusion of Mycobacterium triviale within the M. terrae complex was confirmed. The limited utility of biochemical tests and of mycolic acid analyses for the differentiation of the members of M. terrae complex was also confirmed. The survey allowed the recognition of three previously undescribed species that were characterized by unique sequences in the 16S rRNA, hsp65 and rpoB genes. Mycobacterium engbaekii sp. nov. (proposed previously 40 years ago but never validly published) was characterized by pink photochromogenic pigmentation and rapid growth; phylogenetically it was related to Mycobacterium hiberniae . The type strain of this species, of which eight strains were investigated, is ATCC 27353T ( = DSM 45694T). A cluster of 24 strains was the basis for the description of Mycobacterium heraklionense sp. nov., which has an intermediate growth rate and is unpigmented; nitrate reductase activity is typically strong. Closely related to M. arupense with respect to the 16S rRNA gene, M. heraklionense sp. nov. could be clearly differentiated from the latter species in the other genetic regions investigated. The type strain is NCTC 13432T ( = LMG 24735T = CECT 7509T). Mycobacterium longobardum sp. nov., represented in the study by seven strains, was characterized by a unique phylogenetic location within the M. terrae complex, clearly divergent from any other species. The type strain is DSM 45394T ( = CCUG 58460T).

A Gram-positive-staining, rod-shaped, facultatively oligotrophic bacterial strain, designated MB18T, was isolated from a water sample collected from the River Mahananda at Siliguri (26° 44′ 23.20′ N, 88° 25′ 22.89′ E), West-Bengal, India. On the basis of 16S rRNA gene sequence similarity, the closest relative of this strain was Brevibacterium epidermidis NCDO 2286T (96 % similarity). The DNA G+C content of strain MB18T was 64.6 mol%. Chemotaxonomic data [MK-8(H2) as the major menaquinone, galactose as the sole cell-wall sugar, meso-diaminopimelic acid as the diagnostic cell-wall diamino acid, phosphatidylglycerol and diphosphatidylglycerol as constituents of the polar lipids, anteiso-C15 : 0, anteiso-C17 : 0 and iso-C15 : 0 as the major fatty acids] supported the affiliation of strain MB18T to the genus Brevibacterium . The results of DNA G+C content, 16S rRNA gene sequence analysis and biochemical and physiological analyses allowed genotypic and phenotypic differentiation of strain MB18T from its nearest neighbour B. epidermidis . The isolate therefore represents a novel species, for which the name Brevibacterium siliguriense sp. nov. is proposed; the type strain is MB18T ( = DSM 23676T = LMG 25772T).

A novel Gram-stain-positive rod-shaped actinobacterium was isolated from a soil conditioner made from poultry manure. The isolate, designated strain MN-6-aT, contained anteiso-C15 : 0 and anteiso-C17 : 0 as the major fatty acids, and MK-7(H2) and MK-8(H2) as the major menaquinones. Phosphatidylglycerol was a major polar lipid. The G+C content of the genomic DNA was 67.4 mol%. Phylogenetic analysis showed that strain MN-6-aT was closely related to Brevibacterium salitolerans TRM 415T with 97.1 % 16S rRNA gene sequence similarity. DNA–DNA hybridization showed that strain MN-6-aT had 10.2 % genomic relatedness with B. salitolerans TRM 415T. On the basis of phenotypic, phylogenetic and chemotaxonomic data obtained in this study, strain MN-6-aT represents a novel species of the genus Brevibacterium , for which the name Brevibacterium yomogidense sp. nov. is proposed. The type strain is MN-6-aT ( = JCM 17779T = DSM 24850T).

An actinomycete, designated SA181T, was isolated from Saharan soil in the Hoggar region (south Algeria) and was characterized taxonomically by using a polyphasic approach. The morphological and chemotaxonomic characteristics of the isolate were consistent with the genus Saccharothrix , and 16S rRNA gene sequence analysis confirmed that strain SA181T was a novel member of the genus Saccharothrix . DNA–DNA hybridization values between strain SA181T and its closest phylogenetic neighbours, the type strains of Saccharothrix longispora , Saccharothrix texasensis and Saccharothrix xinjiangensis , were clearly below the 70 % threshold. The genotypic and phenotypic data showed that the isolate represents a novel species of the genus Saccharothrix , for which the name Saccharothrix hoggarensis sp. nov. is proposed, with the type strain SA181T ( = DSM 45457T  = CCUG 60214T).

Strain D10-9-5T was isolated from mangrove soil in Samut Sakhon province, Thailand. A polyphasic approach was used to determine the taxonomic position of the strain. The strain presented single rough spores on substrate mycelium and no aerial mycelium. Chemotaxonomic data supported the assignment of strain D10-9-5T to the genus Micromonospora based on the presence of meso-diaminopimelic acid and glycolyl muramic acid in the peptidoglycan, ribose, mannose, galactose, xylose and glucose as whole-cell sugars, MK-10(H4) (14.8 %), MK-10(H6) (46.7 %) and MK-10(H8) (27.5 %) as the predominant isoprenoid quinones, iso-C15 : 0 (17.9 %), anteiso-C17 : 0 (14.6 %), iso-C17 : 0 (9.6 %), C17 : 0 (8.0 %), iso-C16 : 0 (7.7 %) and C17 : 1ω8c (7.0 %) as the major cellular fatty acids, and diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannosides and phosphatidylethanolamine as the predominant phospholipids in the cell wall. The 16S rRNA gene sequence and phylogenetic analysis showed that strain D10-9-5 was closely related to Micromonospora marina JCM 12870T (99.6 %), Micromonospora coxensis JCM 13248 T (99.4 %), Micromonospora aurantiaca JCM 10878T (99.3 %), Micromonospora humi JCM15292T (99.3 %), Micromonospora halophytica JCM 3125T (99.1%) and Micromonospora chalcea JCM 3031T (99.1 %). Strain D10-9-5T could be clearly distinguished from related members of the genus Micromonospora by its physiological and biochemical characteristics as well as its phylogenetic position and level of DNA–DNA relatedness. Therefore, the strain represents a novel species for which the name Micromonospora maritima sp. nov. is proposed; the type strain is D10-9-5T ( = JCM 17013T = NBRC 108767T = PCU 322T = TISTR 2000T).

An actinomycete strain, designated strain SH2-13T, was isolated from a marine sediment sample collected from the Andaman Sea of Thailand. Applying a polyphasic approach, the isolate was identified as a member of the genus Micromonospora using morphological and chemotaxonomic characteristics, including the presence of meso-diaminopimelic acid in the peptidoglycan. Whole-cell sugars were arabinose, galactose, glucose, rhamnose, ribose and xylose. Diagnostic polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannosides and phosphoglycolipids. The major menaquinones were MK-10(H2), MK-10(H4) and MK-10(H6). 16S rRNA gene sequence analysis revealed similarity to Micromonospora marina JSM1-1T (99.1 %), Micromonospora coxensis 2-30-b(28)T (99.1 %), Micromonospora aurantiaca DSM 43813T (98.8 %) and Micromonospora chalcea DSM 43026T (98.7 %). However, a combination of DNA–DNA hybridization results and phenotypic properties indicated that strain SH2-13T ( = NBRC 107934T = BCC 45601T) should be classified as the type strain of a novel species, with the proposed name Micromonospora sediminicola sp. nov.

The taxonomic position of a soil isolate, strain BK147T, was established using data from a polyphasic study. The organism showed a combination of chemotaxonomic and morphological characteristics consistent with its classification in the genus Actinomadura . It formed a distinct phyletic line in the phylogenetic tree based on 16S rRNA gene sequences of members of the genus Actinomadura and was most closely, albeit loosely, related to Actinomadura bangladeshensis DSM 45347T, Actinomadura meyerae DSM 44715T and Actinomadura napierensis NRRL B-24319T but was readily distinguished from these strains using a range of phenotypic properties. Based on the combined genotypic and phenotypic data it is proposed that isolate BK147T ( = KACC 20919T = NCIMB 14771T = NRRL B-24852T) be classified as the type strain of a novel species of the genus Actinomadura , for which the name Actinomadura xylanilytica sp. nov. is proposed.

A novel actinomycete strain, 7402J-48T, was isolated from an air sample collected from Jeju island, Republic of Korea. Cells were Gram-positive, aerobic, flagellated, short rods. Strain 7402J-48T showed highest 16S rRNA gene sequence similarity (98.6 %) with Angustibacter luteus TT07R-79T, and had relatively low sequence similarities (below 95.1 %) with other members of the family Kineosporiaceae . The cell wall of strain 7402J-48T contained alanine, glutamic acid and 2,6-diaminopimelic acid, suggesting A1γ-type peptidoglycan. The menaquinones were MK-9(H4) and MK-8(H4). The acyl type of the cell-wall muramic acid was acetyl. Diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannoside and an unknown lipid were present. The cellular fatty acid profile comprised a large amount of anteiso-C15 : 0, moderate amounts of C16 : 0, summed feature 3 (including C16 : 1ω7c and/or C16 : 1ω6c), C17 : 0, iso-C15 : 0, C16 : 0 N alcohol and C17 : 1ω8c, and small amounts of other fatty acids. The DNA G+C content of strain 7402J-48T was 73 mol%. Based on its phenotypic and genotypic characteristics, strain 7402J-48T ( = KACC 15527T = NBRC 108730T) is proposed as the type strain of a novel species, Angustibacter aerolatus sp. nov. An emended description of the genus Angustibacter is provided.

A strictly aerobic, Gram-stain-positive actinobacterial strain was isolated from a rhizosphere soil of a wild plant (Peucedanum japonicum Thumb.) collected on Mara Island, Jeju, Republic of Korea. Cells of strain RS-50T were oxidase-negative, catalase-positive, short rods and motile by means of a polar flagellum; the colonies were orange, circular, smooth and convex. meso-Diaminopimelic acid and glucose were the diagnostic diamino acid in the cell wall and the whole-cell sugar, respectively. The major menaquinone was MK-9(H4). The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannoside, two unknown phospholipids and an unknown lipid. The major fatty acids were iso-C15 : 0, iso-C16 : 0, anteiso-C15 : 0 and anteiso-C17 : 0. The DNA G+C content was 73.6 mol%. In 16S rRNA gene sequence-based phylogenetic trees, strain RS-50T formed a tight cluster with Angustibacter luteus (99.2 % sequence similarity); both were loosely related to the suborders Kineosporiineae and Micrococcineae . The DNA–DNA relatedness value of the isolate to A. luteus KACC 14249T was 22.3±0.9 %. On the basis of the results of phenotypic analyses and DNA–DNA hybridization experiments, strain RS-50T is considered to represent a novel species of the genus Angustibacter , for which the name Angustibacter peucedani sp. nov. is proposed. The type strain is RS-50T ( = KCTC 19628T = DSM 45329T). The description of the genus Angustibacter is emended.

Six strains, TKU 25, TKU 28, TKU 30, TKU 31T, TKU 33 and TKU 34, were isolated from the oral cavity of a chimpanzee (Pan troglodytes). Colonies of strains grown on Mitis–Salivarius agar were similar in morphology to that of Streptococcus mutans . The novel strains were Gram-stain-positive, facultatively anaerobic cocci that lacked catalase activity. Analysis of the partial 16S rRNA gene sequences of these isolates showed that the most closely related strain was the type strain of S. mutans (96.4 %). The next closely related strains to the isolates were the type strains of Streptococcus devriesei (94.5 %) and Streptococcus downei (93.9 %). These isolates could be distinguished from S. mutans by inulin fermentation and alkaline phosphatase activity (API ZYM system). The peptidoglycan type of the novel isolates was Glu–Lys–Ala3. Strains were not susceptible to bacitracin. On the basis of phenotypic characterization, partial 16S rRNA gene and two housekeeping gene (groEL and sodA) sequence data, we propose a novel taxon, Streptococcus troglodytae sp. nov.; the type strain is TKU 31T ( = JCM 18038T = DSM 25324T).

A Gram-stain-negative, non-motile, aerobic, endospore forming and rod-shaped bacterium, designated strain L10T, was isolated from marine sediment collected from the South Korean coast. The organism grew optimally under conditions of 30 °C, 1 % (w/v) NaCl and pH 6.0. It was oxidase-negative and catalase-positive. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain L10T was associated with the genus Paenibacillus and most closely related to Paenibacillus barcinonensis BP-23T (98.2 % similarity). The major fatty acids of strain L10T were iso-C14 : 0, anteiso-C15 : 0 and iso-C16 : 0. The cell-wall peptidoglycan was the A1γ type, and the predominant isoprenoid quinone was menaquinone-7. Strain L10T contained two unidentified lipids, an unidentified amino-phospholipid, phosphatidylethanolamine, phosphatidylglycerol, and diphosphatidylglycerol. The G+C content of the genomic DNA was 44 mol% and the DNA–DNA hybridization values with closely related strains were below 14±2 %. Based on phenotypic, genotypic, and phylogenetic data, strain L10T should be classified as a novel species within the genus Paenibacillus . The name Paenibacillus oceanisediminis sp. nov. is proposed. The type strain is L10T ( = KACC 16203T = JCM 17814T).

A facultative anaerobic, non-motile, non-spore-forming, Gram-positive-staining, coccus-shaped bacterium was isolated from an abscess on the right foot of a chimpanzee (Pan troglodytes). The colonies were β-haemolytic. Catalase and oxidase activities were negative. The Lancefield group B antigen was expressed. On the basis of morphological and biochemical characteristics, the bacterium was tentatively identified as a streptococcal species. 16S rRNA gene sequence analysis indicated that the bacterium shared 96.7 %, 96.4 %, 96.1 %, 95.8 % and 95.7 % sequence similarities with Streptococcus gordonii , S. cristatus , S. intermedius , S. anginosus and S. constellatus , respectively. Phylogenetic analyses based on the sequences of the 16S rRNA gene and housekeeping genes encoding d-alanine : d-alanine ligase (ddl), the β-subunit of RNA polymerase (rpoB) and manganese-dependent superoxide dismutase (sodA) revealed that the bacterium represented a novel species closely related to, albeit different from, S. gordonii , S. cristatus and the anginosus streptococci. The name Streptococcus troglodytidis sp. nov. is proposed. The type strain is M09-11185T ( = ATCC BAA-2337T = KCTC 33006T).

Two indigo-reducing alkaliphilic strains, designated strain C40T and strain N214, were isolated from a fermented Polygonum Indigo (Polygonum tinctorium Lour.) liquor sample aged for 10 months and obtained from Date City, Hokkaido, Japan. 16S rRNA gene sequence phylogeny suggested that strains C40T and N214 were members of the genus Amphibacillus with the closest relative being Amphibacillus xylanus JCM 7361T (97.5 % 16S rRNA gene sequence similarity with strain C40T), which is the only strain having a 16S rRNA gene sequence similarity higher than 97 % with strain C40T. Cells of strain C40T were Gram-stain-positive, facultatively anaerobic, straight rods that were motile by means of peritrichous flagella. The strains grew between 17 and 39 °C (optimum, 35 °C) and in the pH range of 9.0–12.0. No isoprenoid quinone was detected and the DNA G+C content was 37.5–37.7 mol%. The whole-cell fatty acid profile mainly consisted of iso-C15 : 0 and anteiso-C15 : 0. DNA–DNA hybridization of strain C40T with Amphibacillus xylanus JCM 7361T revealed a DNA–DNA relatedness value of 10±3 %. Owing to the differences in phenotypic characteristics and phylogenetic analyses based on 16S rRNA gene sequences, as well as DNA–DNA relatedness data from reported species of the genus Amphibacillus , the isolates merit classification as a novel species in the genus Amphibacillus , for which the name Amphibacillus indicireducens sp. nov. is proposed. The type strain is C40T ( = JCM 17250T = NCIMB 14686T). An additional strain of the species is N214. An emended description of the genus Amphibacillus is provided.

A novel thermophilic bacterium, strain Kam1851T, was isolated from a terrestrial hot spring of the Uzon Caldera, Kamchatka Peninsula, Russia. Cells of strain Kam1851T were spore-forming rods with a Gram-positive type of cell wall. Growth was observed between 46 and 78 °C, and pH 5.5–8.5. The optimal growth (doubling time, 6.0 h) was at 60–65 °C and pH 6.5. The isolate was an obligate anaerobe growing in pre-reduced medium only. It grew on mineral medium with molecular hydrogen or formate as electron donors, and elemental sulfur, thiosulfate or polysulfide as electron acceptors. The main cellular fatty acids were C16 : 0 (34.2 %), iso-C16 : 0 (18 %), C18 : 0 (12.8 %) and iso-C17 : 0 (11.1 %). The G+C content of the genomic DNA of strain Kam1851T was 63 mol%. 16S rRNA gene sequence analysis showed that strain Kam1851T belonged to the order Thermoanaerobacterales , but it was not closely related to representatives of any genera with validly published names. The most closely related strains, which had no more than 89.2 % sequence similarity, were members of the genera Ammonifex and Caldanaerobacter . On the basis of its phylogenetic position and novel phenotypic features, isolate Kam1851T is proposed to represent a novel species in a new genus, Brockia lithotrophica gen. nov., sp. nov.; the type strain of Brockia lithotrophica is Kam1851T ( = DSM 22653T = VKM B-2685T).

A set of 25 urease-producing, yellow-pigmented enterococci was isolated from environmental sources. Phenotypic classification divided the isolates into two phena. Both phena were characterized using 16S rRNA gene sequence analysis, DNA base composition, rep-PCR fingerprinting and automated ribotyping. The obtained data distinguished the isolates from all members of the genus Enterococcus with validly published names and placed them in the Enterococcus faecalis species group. DNA–DNA hybridization experiments, pheS and rpoA sequencing and whole-cell protein electrophoresis provided conclusive evidence for the classification of each phenon as a novel species of the genus Enterococcus , for which the names Enterococcus ureilyticus sp. nov. (type strain CCM 4629T  = LMG 26676T  = CCUG 48799T), inhabiting water and plants, and Enterococcus rotai sp. nov. (type strain CCM 4630T  = LMG 26678T  = CCUG 61593T), inhabiting water, insects (mosquitoes) and plants, are proposed.

A study was performed on three isolates (LU2006-1T, LU2006-2 and LU2006-3), which were sampled independently from cheese in western Switzerland in 2006, as well as a fourth isolate (A11-3426), which was detected in 2011, using a polyphasic approach. The isolates could all be assigned to the genus Listeria but not to any known species. Phenotypic and chemotaxonomic data were compatible with the genus Listeria and phylogenetic analysis based on 16S rRNA gene sequences confirmed that the closest relationships were with members of this genus. However, DNA–DNA hybridization demonstrated that the isolates did not belong to any currently described species. Cell-wall-binding domains of Listeria monocytogenes bacteriophage endolysins were able to attach to the isolates, confirming their tight relatedness to the genus Listeria . Although PCR targeting the central portion of the flagellin gene flaA was positive, motility was not observed. The four isolates could not be discriminated by Fourier transform infrared spectroscopy or pulsed-field gel electrophoresis. This suggests that they represent a single species, which seems to be adapted to the environment in a cheese-ripening cellar as it was re-isolated from the same type of Swiss cheese after more than 5 years. Conjugation experiments demonstrated that the isolates harbour a transferable resistance to clindamycin. The isolates did not exhibit haemolysis or show any indication of human pathogenicity or virulence. The four isolates are affiliated with the genus Listeria but can be differentiated from all described members of the genus Listeria and therefore they merit being classified as representatives of a novel species, for which we propose the name Listeria fleischmannii sp. nov.; the type strain is LU2006-1T ( = DSM 24998T  = LMG 26584T).

This study addressed the taxonomic position and group classification of a phytoplasma responsible for virescence and phyllody symptoms in naturally diseased Madagascar periwinkle plants in western Malaysia. Unique regions in the 16S rRNA gene from the Malaysian periwinkle virescence (MaPV) phytoplasma distinguished the phytoplasma from all previously described ‘ Candidatus Phytoplasma ’ species. Pairwise sequence similarity scores, calculated through alignment of full-length 16S rRNA gene sequences, revealed that the MaPV phytoplasma 16S rRNA gene shared 96.5 % or less sequence similarity with that of previously described ‘ Ca. Phytoplasma ’ species, justifying the recognition of the MaPV phytoplasma as a reference strain of a novel taxon, ‘Candidatus Phytoplasma malaysianum’. The 16S rRNA gene F2nR2 fragment from the MaPV phytoplasma exhibited a distinct restriction fragment length polymorphism (RFLP) profile and the pattern similarity coefficient values were lower than 0.85 with representative phytoplasmas classified in any of the 31 previously delineated 16Sr groups; therefore, the MaPV phytoplasma was designated a member of a new 16Sr group, 16SrXXXII. Phytoplasmas affiliated with this novel taxon and the new group included diverse strains infecting periwinkle, coconut palm and oil palm in Malaysia. Three phytoplasmas were characterized as representatives of three distinct subgroups, 16SrXXXII-A, 16SrXXXII-B and 16SrXXXII-C, respectively.