- Volume 56, Issue 4, 2006

The taxonomic position of two mycolic-acid-producing actinomycetes, isolates J81T and J82, which were recovered from activated sludge foam, was clarified. Comparative 16S rRNA gene sequence studies indicated that the organisms formed a distinct lineage within the Corynebacterineae 16S rRNA gene tree. The taxonomic integrity of this group was underpinned by a wealth of phenotypic data, notably characteristic rudimentary right-angled branching. In addition, isolate J81T contained the following: meso-diaminopimelic acid, arabinose and galactose; N-glycolated muramic acid residues; a dihydrogenated menaquinone with eight isoprene units as the predominant isoprenologue; a fatty acid profile rich in oleic and palmitoleic acids and with relatively small proportions of myristic, stearic and tuberculostearic acids; mycolic acids with 44–52 carbons; and diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol and phosphatidylinositol mannosides as major polar lipids. Strain J81T was found to have a chemotaxonomic profile that serves to distinguish it from representatives of all of the other taxa classified as belonging to the suborder Corynebacterineae. In the light of these data, it is proposed that the two isolates be classified in a novel monospecific genus. The name proposed for this taxon is Millisia brevis gen. nov., sp. nov.; strain J81T (=DSM 44463T=NRRL B-24424T) is the type strain of Millisia brevis.

Taxonomic studies were performed on 13 clinical isolates (ten of which were epidemiologically related) that had been previously identified as Oerskovia turbata. Comparative phylogenetic analysis, based on 16S rRNA gene sequences, indicated that the isolates are closely related to Cellulosimicrobium cellulans with sequence similarity values ranging from 99·5 to 99·8 %. Chemotaxonomic results (fatty acid profiles and menaquinones) supported the inclusion of these isolates in the genus Cellulosimicrobium. The DNA G+C content was 74·5 mol%. The results of DNA–DNA reassociation, whole-cell sugars (with galactose as the characteristic whole sugar) and phenotypic properties, including antimicrobial resistance, indicated that these isolates are representatives of a novel species of the genus Cellulosimicrobium. The name Cellulosimicrobium funkei sp. nov. is proposed for the novel strains, with strain W6122T (=ATCC BAA-886T=DSM 16025T=CCUG 50705T) as the type strain. The definition of this novel Cellulosimicrobium species will assist in the understanding of the epidemiology and clinical significance of these micro-organisms.

A taxonomic study was performed on 13 bacterial strains isolated from preputial swabs of European bison (Bison bonasus) bulls suffering from balanoposthitis. The isolates were Gram-positive, non-motile, facultatively anaerobic, diphtheroid-shaped cells. Based on biochemical profiles and BOX-PCR-generated genomic fingerprints, the isolates were grouped into two clusters represented by four and nine strains, respectively. Strains 1(W3/01)T and 2(W106/04)T, selected as representatives of the two clusters, shared 97·2 % 16S rRNA gene sequence similarity. The highest gene sequence similarities found (95·5–96·4 %) were to Arcanobacterium pyogenes DSM 20630T and Arcanobacterium bernardiae DSM 9152T, demonstrating that the novel strains are members of the genus Arcanobacterium, but are not members of a recognized species. The polar lipid profiles of the two novel strains displayed the major characteristics also found in A. pyogenes DSM 20630T and Arcanobacterium haemolyticum DSM 20595T. Detection of a quinone system with MK-10(H4) as the predominant compound confirmed phylogenetic relatedness of the novel strains to A. pyogenes and separated them from the type species of the genus, A. haemolyticum, which contains MK-9(H4) as the predominant quinone. Results from DNA–DNA hybridizations clearly demonstrated that strains 1(W3/01)T and 2(W106/04)T represent separate species. Based on these data, two novel species of the genus Arcanobacterium are described, for which the names Arcanobacterium bialowiezense sp. nov. [type strain 1(W3/01)T=DSM 17162T=NCTC 13354T] and Arcanobacterium bonasi sp. nov. [type strain 2(W106/04)T=DSM 17163T=NCTC 13355T] are proposed.

Strain EJ-46T, a novel pleomorphic, aerobic, extremely halophilic member of the Archaea was isolated from sediment of the saline Lake Ejinor, in Inner Mongolia, China. This organism was neutrophilic and required at least 15 % (2·5 M) NaCl for growth. MgCl2 was not required. The isolate was able to grow at pH 6·0–9·0. Optimum growth occurred in media containing 20 % (3·4 M) NaCl at pH 7·0–7·5. Polar lipid analysis revealed the presence of phosphatidylglycerol and phosphatidylglycerol phosphate methyl ester, derived from both C20C20 and C20C25 glycerol diethers. Four glycolipids were detected, one of which may be novel. The DNA G+C content was 60·3 mol%. 16S rRNA gene analysis revealed that strain EJ-46T was a member of the phylogenetic group defined by the family Halobacteriaceae, and the highest 16S rRNA gene similarity values of 94·9 and 94·8 % were obtained with the haloalkaliphilic species of the genus Natronococcus, Natronococcus occultus and Natronococcus amylolyticus, respectively. Based on the phenotypic, genotypic and phylogenetic analyses, it is proposed that the novel isolate should be classified as representing a new genus and species, for which the name Halovivax asiaticus gen. nov., sp. nov. is proposed. The type strain is EJ-46T (=CGMCC 1.4248T=CECT 7098T).

A marine bacterium, GW1-1T, capable of degrading benzo[a]pyrene (BaP), was isolated from estuarine sediments of the South Sea (the Korea Strait), Korea, after an enrichment culture maintained for 2 years in a medium supplemented with a mixture of BaP and pyrene. The strain formed yellowish-brown colonies on marine agar 2216. Cells were strictly aerobic, non-motile, Gram-negative rods and produced non-diffusible carotenoid pigments. Optimal growth occurred in the presence of 1 % (w/v) NaCl and at pH 7 and 33–36 °C. No growth occurred without supplementation with either CaCl2 or MgCl2, even in the presence of NaCl. Phylogenetic analysis based on the nearly complete sequence of the 16S rRNA gene revealed that the isolate formed a phyletic lineage with the genera Gelidibacter (93·9–94·7 % gene sequence similarity), Subsaximicrobium (93·3 %) and Subsaxibacter (93·9 %). The isolate also showed high sequence similarities to Gaetbulibacter saemankumensis (94·5 %), Algibacter lectus (94·2 %), members of the genus Bizionia (93·6–94·3 %) and Formosa algae (93·2 %), even though it belonged to a different phyletic line. The major respiratory quinones of the isolate were menaquinones MK-5 and MK-6. The DNA G+C content was 51·4 mol%. Dominant fatty acids were i-15 : 0, a-15 : 0, i-15 : 1ω10c and 16 : 1. On the basis of this polyphasic taxonomic evidence, strain GW1-1T is classified as a member of a novel genus and species in the family Flavobacteriaceae, for which the name Yeosuana aromativorans gen. nov., sp. nov. is proposed. The type strain of the type species is GW1-1T (=KCCM 42019T=JCM 12862T).

A rod-shaped marine bacterium, designated strain CL-TF09T, isolated from a tidal flat in Ganghwa, Korea, was characterized based on its physiological and biochemical features, fatty acid profile and phylogenetic position. 16S rRNA gene sequence analysis revealed a clear affiliation with the family Flavobacteriaceae. Strain CL-TF09T showed the closest phylogenetic relationship with the genera Tenacibaculum and Polaribacter; sequence similarities between CL-TF09T and the type strains of Tenacibaculum and Polaribacter species ranged from 90·7 to 91·8 %. Cells of strain CL-TF09T were non-motile and grew on solid media as yellow colonies. The strain grew in the presence of 1–5 % sea salts, within a temperature range of 5–30 °C and at pH 7–8. The strain had iso-C15 : 0 3-OH (17·4 %), iso-C15 : 0 (16·7 %), anteiso-C15 : 0 (15·1 %) and iso-C16 : 0 3-OH (13·4 %) as predominant fatty acids. The DNA G+C content was 33·9 mol%. Based on the physiological, fatty acid composition and phylogenetic data presented, strain CL-TF09T is considered to represent a novel genus and species of the family Flavobacteriaceae, for which the name Lutibacter litoralis gen. nov., sp. nov. is proposed. The type strain is CL-TF09T (=KCCM 42118T=JCM 13034T).

A Gram-negative, non-motile, non-spore-forming bacterial strain, DS-44T, was isolated from soil from Dokdo in Korea, and its taxonomic position was investigated by using a polyphasic approach. It grew optimally at 25 °C and in the presence of 2 % (w/v) NaCl. Strain DS-44T contained MK-7 as the predominant menaquinone and iso-C15 : 0 and C16 : 1 ω7c and/or iso-C15 : 0 2-OH as the major fatty acids. The DNA G+C content was 49·0 mol%. Phylogenetic analyses based on 16S rRNA gene sequences revealed that strain DS-44T belongs to the genus Algoriphagus of the phylum Bacteroidetes. Similarity values between the 16S rRNA gene sequences of strain DS-44T and those of the type strains of recognized Algoriphagus species were in the range 93·8–95·7 %, making it possible to categorize strain DS-44T as a species that is separate from previously described Algoriphagus species. On the basis of its phenotypic properties and phylogenetic distinctiveness, strain DS-44T (=KCTC 12545T=CIP 108837T) was classified in the genus Algoriphagus as the type strain of a novel species, for which the name Algoriphagus terrigena sp. nov. is proposed.

The taxonomic position of four Gram-negative, rod-shaped, golden-yellow-coloured bacteria isolated from marine sediments was determined. Analysis of the almost complete 16S rRNA gene sequences indicated that these isolates belong to the family Flavobacteriaceae. An unclassified bacterium, NBRC 15975, was found to be the closest relative, showing 16S rRNA gene sequence similarity of 93 %; other related genera shared only 87·9–90·5 % similarity. In contrast, the four isolates shared high levels of 16S rRNA gene sequence similarity (99·3–99·7 %) and high DNA–DNA reassociation values (93–104 %). The isolates could be differentiated phenotypically from other genera by the abilities to reduce nitrate and to degrade gelatin, casein and starch. The only respiratory quinone was MK-6, and the major fatty acids were iso-C15 : 0, iso-C15 : 1, anteiso-C15 : 0, iso-C17 : 1 ω9c and iso-C17 : 0 3-OH. The DNA G+C content was 38–40 mol%. Differentiating phenotypic characteristics and large phylogenetic distances between the isolates and previously published genera indicated that the isolates constitute a novel genus, for which the name Sediminicola gen. nov. is proposed. The type species is Sediminicola luteus sp. nov. (type strain CNI-3T=NBRC 100966T=LMG 23246T).

A sponge-associated strain, KMM 7019T, was investigated in a polyphasic taxonomic study. The bacterium was strictly aerobic, heterotrophic, Gram-negative, yellow-pigmented, motile by gliding and oxidase-, catalase-, β-galactosidase- and alkaline phosphatase-positive. A phylogenetic analysis based on 16S rRNA gene sequences revealed that strain KMM 7019T is closely related to members of the genus Salegentibacter, namely Salegentibacter holothuriorum, Salegentibacter mishustinae and Salegentibacter salegens (97·7–98 % sequence similarities). The DNA–DNA relatedness between the strain studied and Salegentibacter species ranged from 27 to 31 %, clearly demonstrating that KMM 7019T belongs to a novel species of the genus Salegentibacter, for which the name Salegentibacter agarivorans sp. nov. is proposed. The type strain is KMM 7019T (=KCTC 12560T=LMG 23205T).

A pale-pink strain (CC-FR2-10T) from the rhizosphere of the sacred tree Ficus religiosa L. in Taiwan was investigated by using a polyphasic taxonomic approach. The cells were Gram-positive, rod-shaped and non-spore-forming. Phylogenetic analyses using the 16S rRNA gene sequence of the isolate indicated that the organism belongs to the genus Deinococcus, the highest sequence similarities being found with Deinococcus grandis (96·1 %), Deinococcus radiodurans (94·3 %), Deinococcus radiopugnans (93·2 %), Deinococcus indicus (93·0 %), Deinococcus proteolyticus (92·5 %), Deinococcus murrayi (92·4 %) and Deinococcus geothermalis (90·7 %). The DNA–DNA relatedness with respect to D. grandis DSM 3963T was 17·9 %. Chemotaxonomic data revealed that strain CC-FR2-10T contains only menaquinone MK-8 as the respiratory quinone, unknown phosphoglycolipids as the predominant polar lipids and 16 : 1ω7c, 17 : 1ω8c and 17 : 1ω9c iso as the predominant fatty acids. The biochemical and chemotaxonomic properties demonstrate that strain CC-FR2-10T represents a novel species, for which the name Deinococcus ficus sp. nov. is proposed. The type strain is CC-FR2-10T (=CCUG 53391T=CIP 108832T).

To evaluate multilocus sequence analysis (MLSA) for taxonomic purposes in the delineation of species within Borrelia burgdorferi sensu lato, seven relevant loci of various strains for which extensive DNA–DNA reassociation data were available were sequenced. MLSA delineation proved to be fully concordant with conventional methods. Our analysis confirmed the delineation of a novel species, Borrelia spielmanii sp. nov., previously known as ‘Borrelia spielmani’ Richter et al. 2004 , with strain PC-Eq17N5T (=DSM 16813T=CIP 108855T) as the type strain.

Strains 18T, 61T and 77 were isolated from two permanently cold fjord sediments on the west coast of Svalbard. The three psychrotolerant strains, with temperature optima at 20–23 °C, were able to grow at the freezing point of sea water, −2 °C. The strains oxidized important fermentation products such as hydrogen, formate and lactate with sulfate as the electron acceptor. Sulfate could be replaced by sulfite, thiosulfate or elemental sulfur. Poorly crystalline and soluble Fe(III) compounds were reduced in sulfate-free medium, but no growth occurred under these conditions. In the absence of electron acceptors, fermentative growth was possible. The pH optimum for the strains was around 7·1. The DNA G+C contents were 43·3 and 42·0 mol% for strains 18T and 61T, respectively. Strains 18T, 61T and 77 were most closely related to Desulfovibrio hydrothermalis (95·0–95·7 % 16S rRNA gene sequence similarity). Strains 18T and 77, exhibiting 99·9 % sequence similarity, represent a novel species for which the name Desulfovibrio frigidus sp. nov. is proposed. The type strain is strain 18T (=DSM 17176T=JCM 12924T). Strain 61T was closely related to strains 18T and 77 (97·6 and 97·5 % 16S rRNA gene sequence similarity), but on the basis of DNA–DNA hybridization strain 61T represents a novel species for which the name Desulfovibrio ferrireducens sp. nov. is proposed. The type strain is strain 61T (=DSM 16995T=JCM 12925T).

Strain T5-04T, a Gram-negative, non-spore-forming, rod-shaped bacterium, was isolated from soil of a ginseng field in South Korea and characterized in order to determine its taxonomic position. 16S rRNA gene sequence analysis revealed that strain T5-04T belongs to the α-4 subgroup of the Proteobacteria, and the highest degrees of sequence similarity determined were to Sphingomonas asaccharolytica IFO 10564T (97·5 %), Sphingomonas koreensis JSS26T (97·1 %), Sphingomonas mali IFO 15500T (96·7 %) and Sphingomonas pruni IFO 15498T (96·6 %). Chemotaxonomic data revealed that strain T5-04T possesses ubiquinone Q-10 predominantly, C18 : 1 as the predominant fatty acid and sphingoglycolipids, all of which corroborate its assignment to the genus Sphingomonas. The results of DNA–DNA hybridization and physiological and biochemical tests clearly demonstrated that strain T5-04T represents a distinct species. Based on polyphasic evidence, T5-04T (=KCTC 12210T=NBRC 100801T=IAM 15213T) should be classified as the type strain of a novel Sphingomonas species, for which the name Sphingomonas soli sp. nov. is proposed.

A novel Pseudomonas species, for which the name Pseudomonas otitidis sp. nov. is proposed, was identified from clinical specimens of infected human ears. Forty-one pseudomonads (34 from patients with acute otitis externa, six from patients with acute otitis media with otorrhoea and one from a patient with chronic suppurative otitis media) were recovered that did not match any known species. On the basis of genetic analyses and biochemical characterization, these isolates were shown to belong to the genus Pseudomonas. 16S rRNA gene sequence analysis and DNA–DNA hybridization studies indicated that this novel bacterium is closely related to, but different from, Pseudomonas aeruginosa. A description of this species is based on polyphasic studies of 11 clinical isolates. The type strain of Pseudomonas otitidis is MCC10330T (=ATCC BAA-1130T=DSM 17224T).

A marine bacterial strain, designated FR1311T, was isolated from a sea-water sample from Ganghwa Island, South Korea. Cells were Gram-negative, facultatively anaerobic, catalase- and oxidase-positive, motile, oval or rod-shaped and halophilic (optimum sea-salt concentration for growth of 5–6 %). Phylogenetic analysis of its 16S rRNA gene sequence revealed that it represented a distinct line of descent within the genus Photobacterium. The major fatty acids were straight-chain saturated (C16 : 0) and monounsaturated fatty acids (C16 : 1 ω7c and C18 : 1 ω7c). The predominant respiratory lipoquinone was Q-8. The DNA G+C content was 44 mol%. The phenotypic features of strain FR1199T were similar to those of Photobacterium damselae subsp. damselae and Photobacterium damselae subsp. piscicida, but several physiological and chemotaxonomic properties readily distinguish the new isolate from them. On the basis of the polyphasic results revealed in this study, FR1311T is considered to be the type strain of a novel species, for which the name Photobacterium ganghwense sp. nov. is proposed. The type strain is FR1311T (=IMSNU 60287T=KCTC 12328T=JCM 12487T).

A Gram-negative, motile and oval- or rod-shaped bacterial strain, DS-43T, was isolated from soil from Dokdo, Korea, and its taxonomic position was investigated by using a polyphasic approach. The strain grew optimally at pH 7·0–8·0 and at 30 °C in the presence of 0·5 % (w/v) NaCl. Strain DS-43T had Q-8 as the predominant ubiquinone and C16 : 0, C16 : 1 ω7c and/or iso-C15 : 0 2-OH, C18 : 1 ω7c and C17 : 0 cyclo as the major fatty acids, which are consistent with the corresponding data for the type strain (KCTC 12459T) of Variovorax paradoxus. The DNA G+C content was 66·0 mol%. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain DS-43T was most closely related to Variovorax paradoxus, the only recognized species of the genus. The level of 16S rRNA gene sequence similarity between strain DS-43T and V. paradoxus IAM 12373T was 97·3 %. DNA–DNA relatedness data and differential phenotypic properties demonstrated that strain DS-43T is distinguishable from V. paradoxus. On the basis of phenotypic, phylogenetic and genetic data, it is proposed that strain DS-43T (=KCTC 12544T=CIP 108838T) be classified in the genus Variovorax as a member of a novel species, Variovorax dokdonensis sp. nov.

Comparative 16S rRNA gene sequence analysis indicates that two distinct sublineages exist within the genus Alcaligenes: the Alcaligenes faecalis lineage, comprising Alcaligenes aquatilis and A. faecalis (with the three subspecies A. faecalis subsp. faecalis, A. faecalis subsp. parafaecalis and A. faecalis subsp. phenolicus), and the Alcaligenes defragrans lineage, comprising A. defragrans. This phylogenetic discrimination is supported by phenotypic and chemotaxonomic differences. It is proposed that the A. defragrans lineage constitutes a distinct genus, for which the name Castellaniella gen. nov. is proposed. The type strain for Castellaniella defragrans gen. nov., comb. nov. is 54PinT (=CCUG 39790T=CIP 105602T=DSM 12141T). Finally, on the basis of data from the literature and new DNA–DNA hybridization and phenotypic data, the novel species Castellaniella denitrificans sp. nov. (type strain NKNTAUT=DSM 11046T=CCUG 39541T) is proposed for two strains previously identified as strains of A. defragrans.

Within a collection of marine strains that were shown to contain the photosynthesis reaction-centre genes pufL and pufM, a novel group of alphaproteobacteria was found and was characterized phenotypically. The 16S rRNA gene sequence data suggested that the strains belonged to the order Rhizobiales and were closest (98·5 % sequence similarity) to the recently described species Hoeflea marina. The cells contained bacteriochlorophyll a and a carotenoid, presumably spheroidenone, in small to medium amounts. Cells of the novel strains were small rods and were motile by means of single polarly inserted flagella. Good growth occurred in complex media with 0·5–7·0 % sea salts, at 25–33 °C (optimum, 31 °C) and at pH values in the range 6–9. With the exception of acetate and malate, organic carbon sources tested supported poor growth or no growth at all. Growth factors were required; these were provided by small amounts of yeast extract, but not by standard vitamin solutions. Growth occurred under aerobic to microaerobic conditions, but not under anaerobic conditions, either in the dark or light. Nitrate was not reduced. Photosynthetic pigments were formed at low to medium salt concentrations, but not at the salt concentration of sea water (3·5 %). On the basis of smaller cell size, different substrate utilization profile and photosynthetic pigment content, the novel strains can be classified as representatives of a second species of Hoeflea, for which the name Hoeflea phototrophica sp. nov. is proposed. The type strain of Hoeflea phototrophica sp. nov. is DFL-43T (=DSM 17068T=NCIMB 14078T).