- Volume 61, Issue 10, 2011

Two novel actinomycete strains, designated KB2-4T and KB9-2, were isolated from soil samples. Both isolates formed finger-shaped sporangia on short sporangiophores that emerged directly from substrate hyphae. The cell-wall peptidoglycan contained glutamic acid, glycine, alanine and meso-diaminopimelic acid; the whole-cell sugars were galactose, glucose, mannose, rhamnose, ribose and xylose. The diagnostic phospholipid was phosphatidylethanolamine. The predominant menaquinones were MK-9(H8) and MK-9(H6). Mycolic acids were not detected. The predominant cellular fatty acids (>20 %) were iso-C16 : 0 and anteiso-C17 : 0. For both strains, the G+C content of the genomic DNA was about 72 mol%. The morphological and chemotaxonomic characteristics of the two strains were typical of members of the genus Dactylosporangium. Phylogenetic analysis using 16S rRNA gene sequences also indicated that these strains should be classified in the genus Dactylosporangium and showed that the closest relative was Dactylosporangium salmoneum NRRL B-16294T (99.1 % 16S rRNA gene sequence similarity). DNA–DNA relatedness and some physiological and biochemical properties indicated that the isolates could be readily distinguished from their closest phylogenetic relatives. On the basis of these phenotypic and genotypic data, the isolates represent a novel species, for which the name Dactylosporangium tropicum sp. nov. is proposed. The type strain is strain KB2-4T ( = BCC 34760T  = JCM 15673T).

A Gram-reaction-positive, non-motile actinobacterium, designated strain 210121T, was isolated from rhizosphere soil of the mangrove fern Acrostichum speciosum. 16S rRNA gene sequence analysis showed that the isolate belonged to the genus Asanoa. DNA–DNA relatedness values between strain 210121T and the type strains of the three recognized species of the genus Asanoa were below the 70 % threshold recommended for distinguishing bacterial genomic species. The novel isolate contained glutamic acid, glycine, alanine and meso-A2pm as cell-wall amino acids, indicating peptidoglycan type A1γ. The characteristic whole-cell sugars were xylose, ribose, glucose and mannose. The predominant menaquinones were MK-9(H4), MK-9(H6) and MK-9(H8). The major fatty acids were iso-C16 : 0 (30.9 %), C17 : 0 (23.0 %), anteiso-C15 : 0 (14.9 %) and iso-C15 : 0 (12.3 %). The phospholipid profile comprised phosphatidylethanolamine, phosphatidylinositol mannosides and phospholipids of unknown structure containing glucosamine. The G+C content of the DNA was 70.3 mol%. On the basis of the phenotypic and genotypic data, strain 210121T ( = CGMCC 4.5593T  = DSM 45427T) represents a novel species of the genus Asanoa, for which the name Asanoa hainanensis sp. nov., is proposed. An emended description of the genus Asanoa is also proposed.

During the course of a study on the bacterial diversity in Western Ghats, India, an actinobacterial strain, designated PC IW02T, was isolated and characterized by a polyphasic taxonomic approach. Strain PC IW02T was a non-motile, Gram-positive, short rod that formed creamish white to yellow coloured colonies. 16S rRNA gene sequence analysis showed that the novel strain showed highest sequence similarity with type strains of members of the genus Dermacoccus: Dermacoccus barathri (96.6 %), Dermacoccus profundi (96.5 %), Dermacoccus abyssi (96.4 %) and Dermacoccus nishinomiyaensis (95.9 %). The phylogenetic tree suggested that strain PC IW02T could represent a member of a new genus of the family Dermacoccaceae with the genus Demetria as closest clade. Pairwise sequence alignment with Demetria terragena HKI 0089T and Kytococcus sedentarius DSM 20547T showed similarities of 94.2 and 93.7 %, respectively. Strain PC IW02T had MK-8(H4) as the major menaquinone. The major fatty acids were iso-C16 : 0 (43.4 %), iso-C16 : 1 H (17.2 %) and anteiso-C17 : 0 (9.9 %). The diagnostic cell-wall amino acid at position 3 of the peptide subunit was lysine; the interpeptide bridge consisted of Gly–Ser–Asp. The polar lipids present were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannosides and phosphatidylserine, along with two unknown phospholipids. The genomic DNA G+C content of the isolate was 77 mol%. On the basis of phenotypic characteristics, including chemotaxonomic data, and 16S rRNA gene sequence similarities, strain PC IW02T represents a novel species in a new genus of the family Dermacoccaceae for which the name Calidifontibacter indicus gen. nov., sp. nov. is proposed. The type strain of Calidifontibacter indicus is PC IW02T ( = MTCC 8338T = DSM 22967T = JCM 16038T). An emended description of the family Dermacoccaceae is provided.

A novel Gram-positive, strictly aerobic and non-motile bacterial strain, S6-4T, was isolated from a Korean alcohol fermentation starter. Optimal growth occurred at 37 °C, at pH 8 and in 1 % (w/v) NaCl. The isolate was positive for oxidase and catalase. It assimilated various sugars and acids were produced from several carbohydrates. The major cell-wall sugars were galactose and arabinose. The major fatty acids of strain S6-4T were C16 : 0, C17 : 1ω9c, C18 : 1ω9c and 10-methyl C18 : 0 (tuberculostearic acid). The predominant isoprenoid quinone was menaquinone MK-9(H2) and peptidoglycan amino acids were meso-diaminopimelic acid, alanine, glycine and glutamic acid. The strain contained mycolic acids. According to phylogenetic analysis based on 16S rRNA gene sequences, strain S6-4T was most closely related to Corynebacterium variabile DSM 20132T (98.1 % similarity). The genomic DNA G+C content of strain S6-4T was 73.6 mol% and DNA–DNA hybridization values with related strains were below 33±4 %. On the basis of phenotypic, genotypic and phylogenetic data, strain S6-4T represents a novel species in the genus Corynebacterium, for which the name Corynebacterium nuruki sp. nov. is proposed; the type strain is S6-4T ( = KACC 15032T  = JCM 17162T).

A Gram-reaction-positive, motile, coccus-shaped actinobacterium, designated strain T2A-S27T, was isolated from a roof tile in Oporto (Portugal) and studied using a polyphasic approach. The 16S rRNA gene sequence of the novel isolate showed high similarity to that of Kineococcus marinus KST3-3T (97.8 % sequence similarity). Strain T2A-S27T showed lower 16S rRNA gene sequence similarities with other members of the genus Kineococcus and members of the family Kineosporiaceae (<94 %). A phylogenetic tree, based on 16S rRNA gene sequences, showed that strain T2A-S27T formed a coherent clade with the type strain of K. marinus and Quadrisphaera granulorum. The isolate was characterized by the presence of meso-diaminopimelic acid in the cell-wall peptidoglycan, MK-9(H2) as the predominant menaquinone and a polar lipid profile consisting of diphosphatidylglycerol and phosphatidylglycerol. The fatty acid profile was dominated by anteiso-C15 : 0. The DNA G+C content was 76.9 mol%. The low level of DNA–DNA relatedness to K. marinus (46–47 %) and the results of the chemotaxonomic and physiological studies clearly distinguished strain T2A-S27T from recognized species of the genus Kineococcus. On the basis of its phylogenetic position and phenotypic traits, strain T2A-S27T ( = LMG 24148T  = CECT 7306T  = DSM 23768T) represents a novel species of a new genus in the family Kineosporiaceae, for which the name Pseudokineococcus lusitanus gen. nov., sp. nov. is proposed. The misclassified species K. marinus is transferred to the new genus as Pseudokineococcus marinus comb. nov. The type strain of Pseudokineococcus marinus is KST3-3T ( = KCCM 42250T  = NRRL B-24439T).

A novel acidothermophilic archaeon, strain Ar-4T, was isolated from a sulfuric hot spring in Tengchong, Yunnan, China. Cells of strain Ar-4T were Gram-staining-negative, irregular cocci and motile by means of flagella. Strain Ar-4T grew over a temperature range of 55–75 °C (optimum, 65 °C), a pH range of 2.5–5.5 (optimum, pH 3.5) and a NaCl concentration range of 0–1 % (w/v). The novel strain was aerobic and facultatively chemolithoautotrophic. The strain could extract metal ions from sulfidic ore. It was also able to oxidize reduced sulfur compounds. In addition, it was able to use heterogeneous organic materials for organotrophic growth. The main cellular lipids were calditoglycerocaldarchaeol (CGTE) and caldarchaeol (DGTE). The DNA G+C content of the strain was 40.2 mol%. Analysis of 16S rRNA gene sequences showed that strain Ar-4T was phylogenetically related to members of the genus Metallosphaera and had sequence similarities of 97.7 %, 97.0 % and 96.8 % with Metallosphaera hakonensis DSM 7519T, Metallosphaera sedula DSM 5348T and Metallosphaera prunae DSM 10039T, respectively. Strain Ar-4T showed DNA–DNA relatedness values of 47.5 %, 30.8 % and 29.1 % with M. hakonensis DSM 7519T, M. sedula DSM 5348T and M. prunae DSM 10039T, respectively. The differences in cell motility, the temperature and pH ranges for growth, the ability to utilize carbon sources, the DNA G+C content, and the low DNA–DNA relatedness values distinguished strain Ar-4T from recognized species of the genus Metallosphaera. On the basis of these results, it was concluded that strain Ar-4T represents a novel species of the genus Metallosphaera, for which the name Metallosphaera cuprina is proposed. The type strain is Ar-4T ( = JCM 15769T = CGMCC 1.7082T).

A methanogenic organism, designated strain HB-1T, from the domain Archaea was isolated from groundwater sampled from a subsurface Miocene formation located in Horonobe, Hokkaido, Japan. The strain grew on methanol, dimethylamine, trimethylamine, dimethylsulfide and acetate but not on monomethylamine, H2/CO2, formate, 2-propanol, 2-butanol or cyclopentanol. Cells were Gram-reaction-negative, non-motile, irregular cocci that were 1.4–2.9 µm in diameter and occurred singly or in pairs. The strain grew at 20–42 °C (optimum 37 °C), at pH 6.0–7.75 (optimum pH 7.0–7.25) and in 0–0.35 M NaCl (optimum 0.1 M). The G+C content of the genomic DNA was 41.4 mol%. 16S rRNA gene sequencing revealed that the strain was a member of the genus Methanosarcina but that it clearly differed from all recognized species of this genus (93.1–97.9 % sequence similarity). The phenotypic and phylogenetic features of strain HB-1T indicate that it represents a novel species of the genus Methanosarcina, for which the name Methanosarcina horonobensis sp. nov. is proposed. The type strain is HB-1T ( = DSM 21571T  = JCM 15518T  = NBRC 102577T).

Two aerobic, Gram-reaction-negative, golden-yellow pigmented and rod-shaped bacteria, designated strains A5Q-118T and A5Q-27, were isolated from an unidentified sea squirt that thrives in the coral reefs off the coast of Okinawa, Japan. Phylogenetic analyses based on the 16S rRNA gene sequence revealed that the novel isolates were affiliated with the family ‘Flammeovirgaceae’ of the phylum Bacteroidetes. Strains A5Q-118T and A5Q-27 shared 100 % sequence similarity with each other and showed <92 % similarity with other cultivated members of the family ‘Flammeovirgaceae’. The novel isolates were phenotypically and physiologically different from strains described previously. The DNA G+C content was 35.5–36.2 mol%, MK-7 was the major menaquinone and iso-C15 : 0 and C16 : 1ω5c were the major fatty acids. Based on the results of this polyphasic taxonomic study, it was concluded that strains A5Q-118T and A5Q-27 represent a novel species in a new genus of the family ‘Flammeovirgaceae’, for which the name Aureibacter tunicatorum gen. nov., sp. nov. is proposed. Proposal for designation of the Flammeovirgaceae fam. nov. is also presented. The type strain of Aureibacter tunicatorum is A5Q-118T ( = KCTC 23232T  = NBRC 107587T).

A Gram-negative, rod-shaped bacterial strain, YC6729T, was isolated from vermicompost collected at Masan, Korea, and its taxonomic position was investigated by a polyphasic taxonomic approach. Strain YC6729T grew optimally at 30 °C and at pH 6.5–8.5. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain YC6729T belongs to the genus Chitinophaga in the family Chitinophagaceae. It was related most closely to Chitinophaga terrae KP01T (96.4 % 16S rRNA gene sequence similarity), Chitinophaga ginsengisegetis Gsoil 040T (96.1 %), Chitinophaga arvensicola IAM 12650T (96.1 %) and Chitinophaga pinensis DSM 2588T (93.3 %). Strain YC6729T contained MK-7 as the major menaquinone and homospermidine as the major polyamine. The fatty acids of strain YC6729T were iso-C15 : 0, C16 : 1ω5c, iso-C17 : 0 3-OH, C16 : 0, anteiso-C18 : 0 and/or C18 : 2ω6,9c, iso-C15 : 0 2-OH and/or C16 : 1ω7c, C14 : 0, iso-C15 : 0 3-OH, iso-C15 : 1 G, C18 : 1ω5c, iso-C15 : 1 I and/or C13 : 0 3-OH, C13 : 0 2-OH, C16 : 0 3-OH and unknown fatty acid ECL 13.565. The polar lipid profile contained phosphatidylethanolamine, unknown aminolipids and unknown lipids. The total DNA G+C content of strain YC6729T was 48.9 mol%. The phenotypic, chemotaxonomic and phylogenetic data showed that strain YC6729T represents a novel species of the genus Chitinophaga, for which the name Chitinophaga eiseniae sp. nov. is proposed. The type strain is YC6729T ( = KACC 13774T  = DSM 22224T).

An orange-coloured, rod-shaped, non-motile, Gram-reaction-negative, strictly aerobic bacterial strain, designated JC2680T, was isolated from a seawater sample of Jeju Island, Korea. The isolate required sea salts for growth. Flexirubin-type pigments were absent. 16S rRNA gene sequence analysis indicated that the test strain belonged to the genus Aquimarina within the family Flavobacteriaceae, but shared relatively low levels of similarity (93.6–95.9 %) with the type strains of recognized Aquimarina species. The predominant cellular fatty acids [iso-C17 : 0 3-OH, iso-C15 : 0 and summed feature 9 (comprising 10-methyl C16 : 0 and/or iso-C17 : 1ω9c) and DNA G+C content (35 mol%) were consistent with the assignment of strain JC2680T to the genus Aquimarina. However, a number of phenotypic characteristics, namely inability to grow under microaerophilic conditions, differences in enzyme reactions, and absence of flexirubin-type pigments and gliding motility, clearly distinguished strain JC2680T from recognized species of the genus Aquimarina. The data presented thus indicate that strain JC2680T represents a novel species of the genus Aquimarina, for which the name Aquimarina addita sp. nov. is proposed. The type strain is JC2680T ( = KACC 14156T  = JCM 17106T).

A bacterial isolate, with an optimum growth temperature of about 50 °C and an optimum pH for growth between 7.5 and 8.5, was recovered from a hot spring in the Furnas area of the Island of São Miguel in the Azores. The novel isolate is orange-pigmented, forms non-motile, rod-shaped cells that stain Gram-negative, is strictly aerobic, oxidase-positive and catalase-negative. The major fatty acids of strain TU-20T are 16 : 1ω6c, 17 : 0 iso 3-OH and 15 : 0 iso. Menaquinone 6 is the major respiratory quinone. Based on 16S rRNA gene sequence analysis, physiological and biochemical characteristics, we describe a novel species of a novel genus represented by strain TU-20T ( = DSM 21410T  = LMG 24594T) for which we propose the name Schleiferia thermophila gen nov., sp. nov. We also propose the family Schleiferiaceae fam. nov. to accommodate this new genus.

A yellow-coloured, rod-shaped, Gram-reaction-negative, aerobic bacterial strain, designated JC2132T, was isolated from a tidal flat sediment sample from Ganghwa Island, Korea. The isolate required sea salts for growth. Cells produced non-diffusible carotenoid pigments, but flexirubin-type pigments were absent. Gliding motility was observed. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain JC2132T represented a distinct phyletic line that reflected a novel generic status within the family Flavobacteriaceae with relatively low sequence similarities (<95 %) to members of other genera with validly published names. The predominant isoprenoid quinone (MK-6) and DNA G+C content (35 mol%) were consistent with assignment of the isolate to the family Flavobacteriaceae, but overall phenotypic traits demonstrated that strain JC2132T was not closely affiliated with any previously described genera. Based on taxonomic data obtained using a polyphasic approach, it is proposed that strain JC2132T represents a novel species in a new genus belonging to the family Flavobacteriaceae, for which the name Snuella lapsa gen. nov., sp. nov. is proposed; the type strain of the type species is JC2132T ( = KACC 14152T = JCM 17111T).

A yellow-coloured, rod-shaped, Gram-reaction-negative and aerobic bacterial strain, designated JC2675T, was isolated from a seawater sample from Jeju Island, Korea. The isolate required sea salts for growth. Gliding motility was observed. Flexirubin-type pigments were absent. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain JC2675T represented a distinct phyletic line that reflected a novel generic status within the family Flavobacteraceae with relatively low gene sequence similarities (<95.7 %) to other recognized genera. The predominant isoprenoid quinone (MK-6) and DNA G+C content (30 mol%) were consistent with the assignment of the novel strain to the family Flavobacteriaceae, but overall phenotypic traits demonstrated that the novel strain was not closely affiliated with any previously described genus. Based on data from a study using a polyphasic taxonomic approach, it is proposed that strain JC2675T represents a new genus and novel species belonging to the family Flavobacteriaceae, for which the name Pontirhabdus pectinivorans gen. nov., sp. nov. is proposed. The type strain of the type species is JC2675T ( = KACC 14153T = JCM 17107T).

A Gram-staining-positive coccus, designated CC-SPL15-2T, was isolated from the rhizosphere of Sesuvium portulacastrum. By 16S rRNA gene sequence analysis, it was shown that strain CC-SPL15-2T belonged to the genus Salinicoccus. The isolate was most closely related to Salinicoccus hispanicus DSM 5352T (98.3 % 16S rRNA gene sequence similarity) and Salinicoccus roseus DSM 5351T (96.7 %); similarities to all other members of the genus Salinicoccus were <96.5 %. In accordance with characteristics of the genus Salinicoccus, the quinone system was mainly composed of menaquinone MK-6. The polar lipid profile exhibited the major components diphosphatidylglycerol, phosphatidylglycerol and an unidentified glycolipid. In the polyamine pattern, spermidine was the predominant compound. The fatty acids were anteiso-C15 : 0, iso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0, which supported the affiliation of strain CC-SPL15-2T to the genus Salinicoccus. DNA–DNA relatedness between strain CC-SPL15-2T and S. hispanicus CCUG 43288T was 42 and 32 % (reciprocal analysis). From these data as well as from physiological and biochemical tests, a clear differentiation of strain CC-SPL15-2T from S. hispanicus and other members of the genus Salinicoccus was possible. We propose that strain CC-SPL15-2T be assigned to a novel species, with the name Salinicoccus sesuvii sp. nov. The type strain is CC-SPL15-2T ( = DSM 23267T  = CCM 7756T).

Five strains (Mizu2-1T, Gon2-7, Koba6-1, Koyu2-2 and Miya2-2) of lactic acid bacteria (LAB) were isolated from flowers in Oze National Park, Japan, using anaerobic cultivation. The five isolates were found to share identical 16S rRNA gene sequences. The isolates exhibited low levels of 16S rRNA gene sequence similarity to known LAB; the closest recognized relatives of strain Mizu2-1T were the type strains of Lactobacillus kunkeei (94.9 %), Lactobacillus kefiri (94.1 %) and Lactobacillus buchneri (93.9 %). Comparative analyses of rpoA and pheS gene sequences demonstrated that the novel isolates did not show significant relationships to other Lactobacillus species. The strains were Gram-stain-positive, catalase-negative and heterofermentative. Anaerobic growth was better than aerobic growth. The isolates utilized a narrow range of carbohydrates as sources of carbon and energy, including glucose and fructose. On the basis of phenotypic characteristics and phylogenetic data, the isolates represent a novel species of the genus Lactobacillus, for which the name Lactobacillus ozensis sp. nov. is proposed. The type strain is Mizu2-1T ( = JCM 17196T  = DSM 23829T).

The development of molecular tools and in particular the use of 16S rRNA gene sequencing has had a profound effect on the taxonomy of many bacterial groups. Gram-positive organisms that encompass the genera Lactobacillus and Clostridium within the Firmicutes are examples of taxa that have undergone major revisions based on phylogenetic information. A consequence of these reorganizations is that a number of organisms are now recognized as being misclassified. Previous studies have demonstrated that Lactobacillus catenaformis and Lactobacillus vitulinus are phylogenetically unrelated to Lactobacillus sensu stricto, being placed within the Clostridia rRNA cluster XVII. Based on the phenotypic, chemotaxonomic and phylogenetic data presented, it is proposed that L. catenaformis and L. vitulinus be reclassified in two new genera, named respectively Eggerthia gen. nov., with the type species Eggerthia catenaformis gen. nov., comb. nov. (type strain DSM 20559T = ATCC 25536T = CCUG 48174T = CIP 104817T = JCM 1121T) and Kandleria gen. nov., with the type species Kandleria vitulina gen. nov., comb. nov. (type strain LMG 18931T = ATCC 27783T = CCUG 32236T = DSM 20405T = JCM 1143T).

An anaerobic, thermophilic bacterium, strain SET IS-9T, was isolated from an Icelandic hot spring. Cells of strain SET IS-9T are short, slightly curved, motile rods. The strain grows chemolithotrophically on CO, producing equimolar quantities of H2 and CO2. It also grows fermentatively on lactate or pyruvate in the presence of yeast extract (0.2 g l−1). Products of pyruvate fermentation are acetate, CO2 and H2. Growth occurs at 50–70 °C, with an optimum at 65 °C, and at pH 5.0–8.0, with an optimum at pH 5.5–6.0. The generation time during chemolithotrophic growth on CO under optimal conditions is 2.0 h. 16S rRNA gene sequence analysis suggested that the organism belongs to the genus Carboxydothermus. On the basis of phenotypic features and phylogenetic analysis, Carboxydothermus islandicus sp. nov. is proposed, with the type strain SET IS-9T ( = DSM 21830T  = VKM B-2561T). An emended description of the genus Carboxydothermus is also given.

An aerobic, saccharolytic, obligately thermophilic, motile, non-spore-forming bacterium, strain T49T, was isolated from geothermally heated soil at Hell’s Gate, Tikitere, New Zealand. On the basis of 16S rRNA gene sequence similarity, T49T is the first representative of a new class in the newly described phylum Armatimonadetes, formerly known as candidate division OP10. Cells of strain T49T stained Gram-negative and were catalase-positive and oxidase-negative. Cells possessed a highly corrugated outer membrane. The major fatty acids were 16 : 0, i17 : 0 and ai17 : 0. The G+C content of the genomic DNA was 54.6 mol%. Strain T49T grew at 50–73 °C with an optimum temperature of 68 °C, and at pH 4.7–5.8 with an optimum growth pH of 5.3. A growth rate of 0.012 h−1 was observed under optimal temperature and pH conditions. The primary respiratory quinone was MK-8. Optimal growth was achieved in the absence of NaCl, although growth was observed at NaCl concentrations as high as 2 % (w/v). Strain T49T was able to utilize mono- and disaccharides such as cellobiose, lactose, mannose and glucose, as well as branched or amorphous polysaccharides such as starch, CM-cellulose, xylan and glycogen, but not highly linear polysaccharides such as crystalline cellulose or cotton. On the basis of its phylogenetic position and phenotypic characteristics, we propose that strain T49T represents a novel bacterial genus and species within the new class Chthonomonadetes classis nov. of the phylum Armatimonadetes. The type strain of Chthonomonas calidirosea gen. nov., sp. nov. is T49T ( = DSM 23976T = ICMP 18418T).