- Volume 60, Issue 6, 2010

A Gram-staining-positive, moderately halophilic bacterium, designated strain Amb31T, was isolated from water of the hypersaline lake Aran-Bidgol in Iran and characterized taxonomically using a polyphasic approach. Cells were rods, motile and able to produce ellipsoidal endospores at a central position in swollen sporangia. Strain Amb31T was facultatively anaerobic and catalase- and oxidase-positive. The strain grew in a complex medium supplemented with 3–25 % (w/v) NaCl (optimum 7.5–10 %). Optimal growth was at 30–35 °C and pH 7.5. Phylogenetic analysis based on 16S rRNA gene sequence comparisons showed that strain Amb31T belonged to the genus Lentibacillus; it exhibited 16S rRNA gene sequence similarity values of 96.8 and 96.4 % to Lentibacillus salicampi SF-20T and Lentibacillus salinarum AHS-1T, respectively, and values of 95.9–94.7 % to the type strains of other recognized species of Lentibacillus. The cell-wall peptidoglycan of strain Amb31T was based on meso-diaminopimelic acid and MK-7 was the respiratory isoprenoid quinone. The major fatty acids were anteiso-C15 : 0 (44.7 %), iso-C16 : 0 (21.4 %) and anteiso-C17 : 0 (15.9 %) and the polar lipid pattern consisted of phosphatidylglycerol, diphosphatidylglycerol, five phospholipids and a glycolipid. The DNA G+C content was 44.1 mol%. All these features confirmed the placement of strain Amb31T within the genus Lentibacillus and the strain could be clearly differentiated from strains of the other species of Lentibacillus on the basis of several phenotypic, genotypic and chemotaxonomic features. DNA–DNA relatedness with the type strain of the most closely related strain, L. salicampi DSM 16425T, was 28 %. Therefore, strain Amb31T represents a novel species of the genus Lentibacillus, for which the name Lentibacillus persicus sp. nov. is proposed. The type strain is Amb31T (=CCM 7683T =CECT 7524T =DSM 22530T =LMG 25304T).

A novel halophilic bacterium, designated strain MSS4T, was isolated from the solar salterns of Mesolongi, Greece. The micro-organism, a motile, Gram-stain-positive, aerobic rod, proliferated at salinities of 1.0–4.0 M NaCl, with optimal growth at 2.5 M NaCl. Endospores were not observed. Strain MSS4T showed optimal growth at 37 °C and pH 8.0. The G+C content of its DNA was 47.2 mol%. The polar lipid pattern of strain MSS4T consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidic acid and phosphatidylethanolamine. It possessed anteiso-C15 : 0, C18 : 0, C16 : 0 and anteiso-C17 : 0 as the major fatty acids (altogether representing 84.7 % of the total). The predominant isoprenoid quinone was MK-7. The cell-wall peptidoglycan contained meso-diaminopimelic acid. 16S rRNA gene sequence analysis showed that the new isolate has 96.1 % similarity to Bacillus qingdaonensis CM1T and Bacillus aidingensis 17-5T, 95.5 % to Bacillus salarius BH169T and lower similarity to other Bacillus species. These results justify the assignment of strain MSS4T to a novel species within the genus Bacillus, for which the name Bacillus halochares sp. nov. is proposed. The type strain is MSS4T (=LMG 24571T =DSM 21373T).

A moderately thermophilic chemoheterotrophic bacterium, strain Mat9-16T, was isolated from microbial mats developed in hot spring water streams from Yumata, Nagano, Japan. Cells of strain Mat9-16T were strictly anaerobic, Gram-stain-negative, non-sporulating, non-motile and short to long rods (2.0–15.5 μm in length). Strain Mat9-16T grew fermentatively with optimum growth at 45 °C, pH 7.0–7.5 and 1 % NaCl (w/v). Phylogenetic analysis based on the 16S rRNA gene revealed that strain Mat9-16T was affiliated with an uncultivated lineage, and the nearest cultivated neighbours were green sulfur bacteria belonging to the class Chlorobea with 77–83 % sequence similarity. However, strain Mat9-16T could not grow phototrophically and did not possess light-harvesting structures, morphologically and genetically, such as the chlorosomes of green sulfur bacteria. On the basis of phenotypic features and phylogenetic position, a novel genus and species are proposed for strain Mat9-16T, to be named Ignavibacterium album gen. nov., sp. nov. (=NBRC 101810T =DSM 19864T). We also propose to place the cultivated bacterial lineage accommodating the sole representative Mat9-16T in a novel class, Ignavibacteria classis nov. In addition, we present a formal description of the phylum-level taxon ‘Chlorobi’ as Chlorobi phyl. nov.

We report the isolation of a novel bacterium, strain C1T, from the midgut of the tsetse fly Glossina palpalis gambiensis, one of the vector insects responsible for transmission of the trypanosomes that cause sleeping sickness in sub-Saharan African countries. Strain C1T is a motile, facultatively anaerobic, rod-like bacterium (0.8–1.0 μm in diameter; 2–6 μm long) that grows as single cells or in chains. Optimum growth occurred at 25–35 °C, at pH 6.7–8.4 and in medium containing 5–20 g NaCl l−1. The bacterium hydrolysed urea and used l-lysine, l-ornithine, citrate, pyruvate, d-glucose, d-mannitol, inositol, d-sorbitol, melibiose, amygdalin, l-arabinose, arbutin, aesculin, d-fructose, d-galactose, glycerol, maltose, d-mannose, raffinose, trehalose and d-xylose; it produced acetoin, reduced nitrate to nitrite and was positive for β-galactosidase and catalase. The DNA G+C content was 53.6 mol%. It was related phylogenetically to members of the genus Serratia, family Enterobacteriaceae, the type strain of Serratia fonticola being its closest relative (99 % similarity between 16S rRNA gene sequences). However, DNA–DNA relatedness between strain C1T and S. fonticola DSM 4576T was only 37.15 %. Therefore, on the basis of morphological, nutritional, physiological and fatty acid analysis and genetic criteria, strain C1T is proposed to be assigned to a novel Serratia species, Serratia glossinae sp. nov. (type strain C1T =DSM 22080T =CCUG 57457T).

An aerobic, Gram-staining-negative, rod or ovoid-shaped bacterial isolate, strain NH52JT, was isolated from a sandy sediment sample from the South China Sea. Strain NH52JT exhibited tumbling motility, formed beige or faint pink colonies, gave a positive reaction in tests for catalase and oxidase and required NaCl for growth. Optimal growth was observed at pH 7.8–9.3, at 30 °C and in the presence of 2.0–4.0 % (w/v) NaCl. The novel strain did not synthesize bacteriochlorophyll a, and the DNA G+C content was 62 %. The major fatty acids were C18 : 1 ω7c, C16 : 0 and C18 : 1 ω7c 11-methyl. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain NH52JT was affiliated to the genus Roseovarius of the class Alphaproteobacteria. Roseovarius pacificus and Roseovarius aestuarii were the most closely related recognized species to strain NH52JT with 16S rRNA gene sequence similarity values of 95.0 and 95.7 %, respectively. Sequence similarity values between strain NH52JT and other phylogenetically related species were all below 95.0 %. Based on the phenotypic, chemotaxonomic and phylogenetic data presented, strain NH52JT is considered to represent a novel species of the genus Roseovarius, for which the name Roseovarius nanhaiticus sp. nov. is proposed. The type strain is NH52JT (=LMG 24840T=CCTCC AB 208317T=MCCC 1A03543T).

A Gram-negative, non-pigmented, non-motile, chemoheterotrophic marine bacterium, designated strain IMCC2782T, was isolated from a surface seawater sample of the Yellow Sea, Korea. On the basis of 16S rRNA gene sequence analysis and phenotypic and genotypic characterization, strain IMCC2782T is shown to represent a novel species of the genus Litoricola. Strain IMCC2782T grew at 15–37 °C and tolerated 7.5 % NaCl. Based on 16S rRNA gene sequences, strain IMCC2782T was related most closely to the type strain of Litoricola lipolytica (97.1 % similarity) but showed less than 90 % similarity to the type strains of other bacterial species. The predominant fatty acids were mono-unsaturated C18 : 1 and C16 : 1. The G+C content of the DNA of strain IMCC2782T was 59.6 mol%. A low level of DNA–DNA relatedness (28.1 %) together with several phenotypic characteristics, including enzyme activities, served to differentiate strain IMCC2782T from the type strain of L. lipolytica. Therefore, strain IMCC2782T is considered to represent a novel species of the genus Litoricola, for which the name Litoricola marina sp. nov. is proposed. The type strain is IMCC2782T (=KCTC 22683T =NBRC 105824T).

A chemo-organotrophic, aerobic, facultatively anaerobic, non-motile strain, MWH-C5T, isolated from the water column of the oligomesotrophic Lake Mondsee (Austria), was characterized phenotypically, phylogenetically and chemotaxonomically. The predominant fatty acids of the strain were C16 : 1 ω7c/ω6c, C16 : 0, C12 : 1 and C8 : 0-3OH, the major quinone was ubiquinone Q-8 and the G+C content of the DNA of the strain was 55.5 mol%. 16S rRNA gene similarity to the closest related type strains was 96.6 % (Curvibacter delicatus LMG 4328T) and 95.7 % (Rhodoferax fermentans FR3T). Phylogenetic analysis of 16S rRNA gene sequences revealed the affiliation of the strain with the family Comamonadaceae (Betaproteobacteria); however, the phylogenetic position of the strain did not support an affiliation to any previously described genus within this family. A family-wide comparison of traits revealed that the strain possesses a unique combination of DNA G+C content, major fatty acids and major 3-hydroxy fatty acid. Furthermore, the strain differs in several traits from the closest related genera. Based on the phylogeny of the strain and differences from closely related genera, we propose to establish the new genus and species Limnohabitans curvus gen. nov., sp. nov. to accommodate this strain. The type strain of Limnohabitans curvus is MWH-C5T (=DSM 21645T =CCUG 56720T). The type strain is closely related to a large number of uncultured bacteria detected by cultivation-independent methods in various freshwater systems.

A Gram-negative, non-motile, pale-yellow, rod-shaped bacterial strain, DS-42T, was isolated from a soil in Korea and its taxonomic position was investigated by a polyphasic study. Strain DS-42T grew optimally at 25 °C and pH 7.0–8.0. Strain DS-42T did not form nodules on three different legumes, and the nodD and nifH genes were also not detected by PCR. Strain DS-42T contained Q-10 as the predominant ubiquinone. The major cellular fatty acid was C18 : 1 ω7c. The DNA G+C content was 60.8 mol%. Phylogenetic analyses based on 16S rRNA, atpD and recA gene sequences showed that strain DS-42T belonged to the genus Rhizobium. Strain DS-42T showed 16S rRNA gene sequence similarity of 94.1–97.7 % to the type strains of recognized Rhizobium species. DNA–DNA relatedness between strain DS-42T and the type strains of Rhizobium huautlense, R. galegae, R. loessense and R. cellulosilyticum was 13–19 %, indicating that strain DS-42T was distinct from them genetically. Strain DS-42T can also be differentiated from these four phylogenetically related Rhizobium species by various phenotypic properties. On the basis of phenotypic properties, phylogenetic distinctiveness and genetic data, strain DS-42T is considered to represent a novel species of the genus Rhizobium, for which the name Rhizobium soli sp. nov. is proposed. The type strain is DS-42T (=KCTC 12873T =JCM 14591T).

A novel betaproteobacterium, designated strain Ch07T, was isolated from granules from the wastewater-treatment bioreactor of an alcohol fermentation factory in South Korea. In order to determine its taxonomic position, the novel strain was characterized using a polyphasic approach. The new strain was Gram-negative, facultatively anaerobic, non-spore-forming, motile and short rod-shaped. 16S rRNA gene sequence analysis revealed that strain Ch07T belonged to the class Betaproteobacteria, being related to Pusillimonas noertemannii BN9T (gene sequence similarity 97.30 %), Achromobacter xylosoxidans subsp. xylosoxidans DSM 10346T (97.09 %), Bordetella pertussis DSM 5571T (97.01 %), Pigmentiphaga kullae DSM 13608T (96.68 %) and Castellaniella defragrans DSM 1214T (96.47 %). The results of DNA–DNA hybridization tests showed that reassociation values were less than 62 % with respect to these closely related type strains. Chemotaxonomic data showed that strain Ch07T possessed ubiquinone Q-8. The G+C content of the genomic DNA was 67.9±0.1 mol%. The major polyamine of strain Ch07T was putrescine. The major polar lipids of strain Ch07T were phosphatidylethanolamine, followed by diphosphatidylglycerol and phosphatidylglycerol. When strain Ch07T was incubated on tryptic soy agar, the major cellular fatty acids were C16 : 0, C17 : 0 cyclo, summed feature 3 (C16 : 1 ω7c/iso C15 : 0 2-OH) and summed feature 5 (C18 : 1 ω7c/ω9t/ω12t). The results of DNA–DNA hybridizations, in combination with the chemotaxonomic and physiological data, demonstrated that strain Ch07T represents a novel species of a new genus, for which the name Parapusillimonas granuli gen. nov., sp. nov. is proposed. The type strain of the type species is Ch07T (=KCTC 12668T=LMG 24012T).

A novel Gram-negative, aerobic, orange-pigmented bacterial strain, designated K7-2T, was isolated from seawater of Gangjin Bay, Korea, and subjected to a polyphasic taxonomic study. Strain K7-2T contained ubiquinone-10 (Q-10) as the predominant respiratory lipoquinone and did not produce bacteriochlorophyll a. Major fatty acids were C18 : 1 ω7c (51.4 %), iso-C15 : 0 2-OH and/or C16 : 1 ω7c (15.0 %) and C17 : 1 ω6c (8.8 %). Major polar lipids were phosphatidylethanolamine and phosphatidylcholine. The DNA G+C content was 61.6 mol%. Phylogenetic analyses based on 16S rRNA gene sequences revealed that strain K7-2T formed a distinct phylogenetic lineage within the cluster comprising Erythrobacter strains. Similarities between the 16S rRNA gene sequences of strain K7-2T and the type strains of Erythrobacter species ranged from 95.0 % (Erythrobacter litoralis DSM 8509T) to 96.8 % (Erythrobacter citreus RE35F/1T). On the basis of polyphasic taxonomic data, strain K7-2T (=KCTC 22330T=JCM 15420T) is classified in a novel species within the genus Erythrobacter, for which the name Erythrobacter gangjinensis sp. nov. is proposed.

Resequencing of the 16S rRNA gene of the type strain of Herbaspirillum putei Ding and Yokota 2004 revealed 99.9 % sequence similarity to that of the type strain of Herbaspirillum huttiense (Leifson 1962) Ding and Yokota 2004. This high phylogenetic relatedness of H. putei and H. huttiense was confirmed by the results of DNA–DNA hybridization between H. huttiense DSM 10281T and H. putei ATCC BAA-806T (reassociation value 96 %). Therefore, it is proposed to reclassify the type strain of H. putei as a strain of H. huttiense. However, the genome of the type strain of H. putei is about 0.9 Mb larger than that of the H. huttiense type strain. This results in a decrease in the reassociation value in the reciprocal DNA–DNA hybridization to 72 %, a level slightly above the threshold for delineating bacterial species. These data and distinctive phenotypic characteristics indicate that the name Herbaspirillum putei is a later heterotypic synonym of Herbaspirillum huttiense and permit the description of two novel subspecies, Herbaspirillum huttiense subsp. huttiense subsp. nov. (type strain ATCC 14670T =JCM 21423T =DSM 10281T) and Herbaspirillum huttiense subsp. putei subsp. nov., comb. nov. (type strain 7-2T =JCM 21495T =ATCC BAA-806T). Three bacterial strains, IEH 4430T, IEH 4515 and IEH 8757, isolated from water were found to be the closest relatives of these strains. Strain IEH 8757 was classified as a strain of H. huttiense subsp. putei. Studies of genotypic and phenotypic features of strains IEH 4430T and IEH 4515 showed that the strains represent a novel species, which is most closely related to H. huttiense and for which the name Herbaspirillum aquaticum sp. nov. is proposed (type strain IEH 4430T =DSM 21191T =ATCC BAA-1628T).

A Gram-stain-negative, rod-shaped, oxidase-positive, non-spore-forming bacterium (Sa15T) was isolated from the air in a duck barn. On the basis of 16S rRNA gene sequence similarity studies, the organism was grouped into the class gammaproteobacteria in the neighbourhood of the genus Fulvimonas. The quinone system consisted exclusively of ubiquinone Q-8. The polar lipid profile was mainly composed of the major lipids diphosphatidylglycerol, phosphatidylmonomethylethanolamine and phosphatidylcholine, and moderate amounts of phosphatidylglycerol and an unidentified lipid. This profile was substantially different from that of Fulvimonas soli LMG 19981T examined concurrently. The polyamine pattern showed the predominant amine spermidine. Major fatty acids (iso-C15 : 0, iso-C17 : 1 ω9c and iso-C17 : 0) were in agreement with its phylogenetic affiliation in the vicinity of Fulvimonas; however, differences in the polar lipid and fatty acid patterns and the polyamine profiles could be observed as well. On the basis of DNA–DNA pairing results, chemotaxonomic data and physiological and biochemical data, the strain can be clearly differentiated from Fulvimonas soli. It is evident that this organism represents a novel genus, for which the name Pseudofulvimonas gallinarii gen. nov., sp. nov. is proposed, with the type strain Sa15T (=DSM 21944T =CCM 7599T).

A Gram-negative, aerobic, mesophilic, non-spore-forming, chemotrophic, chlorophyll-lacking, nitrogen-fixing bacterium, designated strain USBA 355T, was isolated from the saline spring ‘Salado de Consotá’ situated in the Colombian Andes. The non-flagellated cells of strain USBA 355T were straight to slightly curved rods (0.6–0.7 × 3.0–3.5 μm). Growth occurred optimally at 30 °C (growth temperature range between 20 and 40 °C), at pH 6.5–6.7 (pH growth range between 5.0 and 8.0) and at 0.5 % NaCl (w/v) (range between 0 and 4 %). The major quinone present was Q-10 and the predominant fatty acids identified were C19 : 0 cyclo ω8c, C18 : 1 ω7c and C18 : 0. The G+C content of the chromosomal DNA was 71±1 mol%. 16S rRNA gene sequence analysis indicated that strain USBA 355T formed a distant phylogenetic line of descent with members of the genus Thalassobaculum, family Rhodospirillaceae, class Alphaproteobacteria (90 % gene sequence similarity). Comparison of the phylogenetic, chemotaxonomic and physiological features of strain USBA 355T with all other members of the family Rhodospirillaceae suggested that it represents a novel genus and species for which the name Tistlia consotensis gen. nov., sp. nov. is proposed. The type strain of the type species is USBA 355T (=JCM 15529T=KCTC 22406T).

A bacterial strain, designated Nsw-4T, was isolated from a water sample of Niao-Song Wetland Park in Taiwan and was characterized by using a polyphasic taxonomic approach. Strain Nsw-4T was Gram-negative, aerobic, ivory-coloured, rod-shaped and motile by means of a polar flagellum. Growth occurred at 15–37 °C, pH 6.0–8.0 and 0–2 % NaCl. Phylogenetic analyses based on 16S rRNA gene sequences showed that the strain belonged to the genus Deefgea and that its closest neighbour was Deefgea rivuli WB 3.4-79T (96.9 %). The results of physiological and biochemical tests allowed the clear phenotypic differentiation of this isolate from D. rivuli WB 3.4-79T. The major fatty acids were C16 : 1ω7c and C16 : 0. The G+C content of the genomic DNA was 53.7 mol%. On the basis of 16S rRNA gene sequence analysis and the chemotaxonomic and physiological data, strain Nsw-4T should be classified as representing a novel species and the second member of the genus Deefgea, for which the name Deefgea chitinilytica sp. nov. is proposed. The type strain is Nsw-4T (=BCRC 17934T=LMG 24817T).

Thirty-nine yeast strains were recovered from a field nest of a primitive and putative novel attine ant species in the genus Myrmicocrypta (Hymenoptera: Formicidae: tribe Attini). Yeasts isolated from the fungus garden and waste deposit included Candida dubliniensis, Candida oleophila, Cryptococcus haglerorum and Hanseniaspora uvarum. In addition, one morphological type was isolated overwhelmingly. Sequencing data of partial large-subunit (LSU) rDNA and the internal transcribed spacer (ITS) region coupled with morphological and physiological characterization accommodated this morphotype in a separate taxonomic position in relation to the known species of Trichosporon (Basidiomycota: Trichosporonales). Here, we propose a novel yeast species named Trichosporon chiarellii sp. nov. based on the description of 34 isolates; the type strain is strain FCP 540806T (=CBS 11177T).

During an investigation of the microbiology of Ghanaian cocoa fermentations, a number of yeast isolates with unusual pheno- and genotypic properties representing three possible novel species were isolated. Members of Group A divided by multilateral budding and ascospores were not produced. Group B strains produced true hyphae and ascospores were not produced. Group C representatives divided by budding and formed chains and star-like aggregates. Ascospores were not produced. Sequence analysis of the 26S rRNA gene (D1/D2 region) revealed that the Group A isolates were phylogenetically most closely related to Saturnispora mendoncae (gene sequence similarity 92.4 %), Saturnispora besseyi (88.8 %), Saturnispora saitoi (88.8 %) and Saturnispora ahearnii (88.3 %). Members of Group B were most closely related to representatives of the genera Dipodascus and Galactomyces and the asporogenous genus Geotrichum, but in all cases with 26S rRNA gene (D1/D2 region) similarities below 87 %. For Group C, the most closely related species were Candida rugopelliculosa (92.4 %), Pichia occidentalis (91.6 %) and Pichia exigua (91.9 %). The very low gene sequence similarities obtained for the three groups of isolates clearly indicated that they represented novel species. Repetitive Palindromic PCR (Rep-PCR) of the isolates and their closest phylogenetic relatives confirmed that the new isolates belonged to previously undescribed species. In conclusion, based on the genetic and phenotypic results, the new isolates were considered to represent three novel species, for which the names Candida halmiae (group A, type strain G3T=CBS 11009T=CCUG 56721T); Geotrichum ghanense (group B, type strain G6T=CBS 11010T=CCUG 56722T) and Candida awuaii (group C, type strain G15T=CBS 11011T=CCUG 56723T) are proposed.

During a survey of the culturable soil fungal population in samples collected in Taylor Valley, South Victoria Land, Antarctica, 13 basidiomycetous yeast strains with orange-coloured colonies were isolated. Phylogenetic analyses of internal transcribed spacer (ITS) and partial LSU rRNA gene sequences showed that the strains belong to the Dioszegia clade of the Tremellales (Tremellomycetes, Agaricomycotina), but did not correspond to any of the hitherto recognized species. Two novel species, Dioszegia antarctica sp. nov. (type strain ANT-03-116T =CBS 10920T =PYCC 5970T) and Dioszegia cryoxerica sp. nov. (type strain ANT-03-071T =CBS 10919T =PYCC 5967T), are described to accommodate ten and three of these strains, respectively. Analysis of ITS sequences demonstrated intrastrain sequence heterogeneity in D. cryoxerica. The latter species is also notable for producing true hyphae with clamp connections and haustoria. However, no sexual structures were observed. The two novel species can be considered obligate psychrophiles, since they failed to grow above 20 °C and grew best between 10 and 15 °C.

Three teleomorphic ascomycetous yeast isolates (TA11TR-1T, TA11TR-4 and TA11TR-6) from orchard soil from Tai'an, Shandong province, China, were shown to represent a novel species within the genus Kazachstania based on phenotypic characterization and sequence analyses of the 18S rRNA gene, internal transcribed spacer (ITS) regions and 26S rDNA gene D1/D2 domain. The name Kazachstania taianensis sp. nov. (type strain TA11TR-1T =AS 2.4160T =CBS 11405T) is proposed. K. taianensis sp. nov. clustered in a branch together with Kazachstania sinensis, Kazachstania naganishii and the Kazachstania telluris complex with moderate bootstrap support in the neighbour-joining tree reconstructed from combined 18S and D1/D2 sequences. The novel species possessed unusual ITS 1 (338 bp) and ITS 2 (488 bp) sequences. The total length of the ITS–5.8S rDNA gene region of the species was 983 bp, being much longer than those of other ascomycetous yeast species described so far.