- Volume 56, Issue 3, 2006

A xylanolytic bacterium, US15T, was isolated from swamp forest soil in Ulsan, Korea. The cells of the novel strain were Gram-positive, non-motile, short-rod-shaped and showed chemotaxonomic properties that were consistent with its classification in the genus Microbacterium. Chemotaxonomic results showed MK-12 and MK-11 as major menaquinones, predominating iso- and anteiso-branched cellular fatty acids, glucose, galactose and mannose as cell-wall sugars, peptidoglycan-type B2β with glycolyl residues and a DNA G+C content of 66·5 mol%. Phylogenetic analysis based on 16S rRNA gene sequencing showed that strain US15T was closely related to Microbacterium arborescens IFO 3750T, Microbacterium imperiale IFO 12610T and Microbacterium ulmi LMG 20991T (96·9, 96·8 and 96·2 % similarities, respectively), and formed a separate lineage within the genus Microbacterium. Combined genotypic and phenotypic data showed that strain US15T (=DSM 16915T=KCTC 19080T) merits recognition as the type strain of a novel species within the genus Microbacterium, for which the name Microbacterium paludicola sp. nov. is proposed.

Two actinomycete strains, designated N7-3T and N4-6T, were isolated from a natural cave on Jeju Island, Republic of Korea, by using a dilution method, and were subjected to physiological, chemical and molecular characterization. The nearly complete sequences of the 16S rRNA gene were aligned and compared with those of representatives of the genus Amycolatopsis. Phylogenetic analysis showed that the organisms belong to the family Pseudonocardiaceae and formed two distinct lineages within the evolutionary radius of the genus Amycolatopsis. The chemotaxonomic and morphological properties support their classification in the genus Amycolatopsis. The 16S rRNA gene sequence data revealed that the closest relatives of strains N7-3T and N4-6T were Amycolatopsis sulphurea (97·9 % similarity) and Amycolatopsis albidoflavus (98·7 % similarity), respectively. The combination of physiological and genetic data supported the observation that the organisms could be distinguished from each other and from established species of the genus Amycolatopsis. The names Amycolatopsis jejuensis sp. nov. and Amycolatopsis halotolerans sp. nov. are proposed for the two novel species, with N7-3T (=NRRL B-24427T=JCM 13280T) and N4-6T (=NRRL B-24428T=JCM 13279T) as the respective type strains.

Actinomycete strain H9T was isolated from coastal sediment of the Jiao-Dong peninsula (near Tsingdao city) in China, and was identified by means of polyphasic taxonomy. The strain grew autotrophically on modified nitrifying medium and heterotrophically on Luria–Bertani medium, with NaCl ranging from 0 to 8 % (w/v) (optimal growth at 3·5 %). The 16S rRNA gene sequence similarities of strain H9T to members of the genus Pseudonocardia ranged from 93·0 to 97·5 %, indicating that strain H9T was phylogenetically related to members of the genus Pseudonocardia. Strain H9T had type IV cell wall and type PIII phospholipid, and its major menaquinone was MK-8 (H4). DNA–DNA relatedness values between strain H9T and Pseudonocardia kongjuensis, Pseudonocardia autotrophica and Pseudonocardia compacta were 42, 13 and 11 %, respectively. These results support the conclusion that H9T represents a novel species within the genus Pseudonocardia, for which the name Pseudonocardia ammonioxydans sp. nov. is proposed, with the type strain H9T (=CGMCC 4.1877T=JCM 12462T).

A novel actinomycete, strain N3-2T, was isolated from a natural cave on Jeju Island, Republic of Korea, using a dilution method and was subjected to polyphasic taxonomy. The almost complete 16S rRNA gene sequence was determined by direct sequencing of the purified PCR product and was compared with those of representatives of the genus Nocardia. It was revealed from the phylogenetic analysis that the organism forms a distinct clade between the Nocardia salmonicida cluster and the Nocardia alba branch within the evolutionary radius occupied by the genus Nocardia of the family Nocardiaceae. The organism showed 16S rRNA gene sequence similarity of 97·4 % with its nearest phylogenetic neighbours, namely N. salmonicida and N. alba. The chemotaxonomic properties, such as the principal amino acid of peptidoglycan, predominant menaquinone and polar lipids, supported the classification in the genus Nocardia. The organism was readily differentiated from Nocardia species with validly published names by a broad set of phenotypic properties and its unique phylogenetic position; the name Nocardia jejuensis sp. nov. is proposed, with N3-2T (=JCM 13281T=NRRL B-24430T) as the type strain.

The taxonomic status of a bacterium isolated from a perianal swab of a patient with a bone marrow transplant was characterized using a polyphasic taxonomic approach. Chemotaxonomic investigations revealed the presence of cell wall chemotype IV, short chain mycolic acids that co-migrated with those extracted from members of the genus Dietzia, and a dihydrogenated menaquinone with eight isoprene units as the predominant menaquinone. Generic assignment was confirmed by 16S rRNA gene sequencing. Comparative analysis of the 16S rRNA gene sequence showed that this isolate constitutes a distinct phyletic line within the genus Dietzia, displaying 97·5–98·7 % sequence similarity with Dietzia species with validly published names. The isolate could be distinguished from the type strain of Dietzia maris (1·6 % sequence divergence) and other species of the genus Dietzia by DNA–DNA hybridization, as well as by using a set of biochemical tests. Genotypic and phenotypic data show that the strain merits classification as a novel species of the genus Dietzia for which the name Dietzia cinnamea sp. nov. is proposed; the type strain is IMMIB RIV-399T (=DSM 44904T=CCUG 50875T).

A yellow-pigmented, non-motile, Gram-negative bacterium, designated Fg 69T, was isolated from a sediment sample collected in Chazhma Bay (Sea of Japan). The novel organism grew at 10–35 °C, was neutrophilic and required 3–10 % NaCl for optimal growth. Strain Fg 69T was able to degrade starch and to hydrolyse gelatin and Tween 80 weakly but not casein or agar. Predominant cellular fatty acids comprised n-C15 and n-C16 branched-chain and straight-chain saturated and unsaturated fatty acids, including iso-C15 : 0 (5 %), anteiso-C15 : 0 (11 %), C15 : 0 (9 %), iso-C15 : 1 (5 %), iso-C16 : 0 (8 %), C16 : 0 (5 %) and C16 : 1 ω7 (5 %) and iso- and anteiso-branched 2-OH and 3-OH C15 : 0 to C17 : 0 fatty acids (26 % in total). The G+C content of the DNA was 40·4 mol%. 16S rRNA gene sequence data indicated that strain Fg 69T belonged to the genus Salegentibacter but was distinct from recognized Salegentibacter species (94–95 % sequence similarity). Based on these results, a novel species, Salegentibacter flavus sp. nov., is proposed. The type strain is Fg 69T (=KMM 6000T=CIP 107843T).

Eight strains of anaerobic Gram-negative bacilli isolated from infections of the skin and soft tissues were subjected to a comprehensive range of phenotypic and genotypic tests. 16S rRNA gene sequence analysis revealed the strains to constitute a homogeneous group, distinct from species with validly published names but related to a cluster including Prevotella buccae, Prevotella dentalis and Prevotella baroniae. A novel species, Prevotella bergensis sp. nov., is proposed to accommodate these strains. Prevotella bergensis is saccharolytic and produces acetic and succinic acids as end products of fermentation. The G+C content of the DNA of the type strain is 48 mol%. The type strain of Prevotella bergensis is 94067913T (=DSM 17361T=CCUG 51224T).

A rod-shaped bacterium, designated CL-TF13T, was isolated from a tidal flat in Ganghwa, Korea. Analysis of the 16S rRNA gene sequence revealed an affiliation with the genus Tenacibaculum. The sequence similarities between CL-TF13T and type strains of members of the genus Tenacibaculum were from 94·2 to 97·4 %. Cells were motile by means of gliding. Strain CL-TF13T grew on solid medium as pale-yellow colonies with an irregular spreading edge. The strain was able to grow in NaCl at a range of 3–5 %. They grew within a temperature range of 5–40 °C and at pH range of 6–10. The major fatty acids were summed feature 3 (C16 : 1 ω7c and/or iso-C15 : 0 2-OH, 19·6 %), iso-C15 : 0 (18·8 %) and iso-C17 : 0 3-OH (13·6 %). Fatty acids such as C18 : 3 ω6c (6,9,12) (1·5 %) and summed feature 4 (iso I- and/or anteiso B-C17 : 1, 1·3 %) were uniquely found in minor quantities in CL-TF13T among Tenacibaculum species. The DNA G+C content was 30 mol%. According to physiological data, fatty-acid composition and 16S rRNA gene sequence, CL-TF13T could be assigned to the genus Tenacibaculum but distinguished from the recognized species of the genus. Therefore, strain CL-TF13T (=KCCM 42115T=JCM 13039T) represents a novel species, for which the name Tenacibaculum litoreum sp. nov. is proposed.

The prevalence of leptospirosis is generally high in domestic animals and rodents in Tanzania. Identification of Leptospira isolates from cattle was carried out to establish prevalent Leptospira serovars. Serological typing was done based on monoclonal antibodies and the standard cross-agglutination absorption test. Molecular typing involved pathogenic- and saprophytic-specific PCRs and a PCR specifically amplifying DNA from the species Leptospira kirschneri. DNA fingerprinting with primers derived from sequences of insertion elements IS1500 and IS1533 was carried out. Both serological and molecular characterization indicated that one of the Leptospira isolates, coded RM1, represents a new serovar of the species L. kirschneri of serogroup Icterohaemorrhagiae. The serovar name Sokoine is proposed for this new Leptospira isolate.

Isolates of Leptospira from two human cases of leptospirosis in Denmark and France were studied using DNA–DNA relatedness, G+C content, 16S rRNA gene sequence data and pulsed-field gel electrophoresis. These isolates differed from previously described species of Leptospira and are defined as Leptospira broomii sp. nov. The type strain is 5399T (=ATCC BAA-1107T=KIT 5399T).

Phylogenetic and phenotypic analysis of cultivable marine bacteria isolated from laboratory cultures of two paralytic shellfish toxin-producing dinoflagellates, Gymnodinium catenatum and Alexandrium tamarense, showed the presence of a novel group of Gram-negative, aerobic, moderately halophilic and hydrocarbon-degrading bacteria, related to the genus Marinobacter. The strains, designated DG893T, DG1136 and ATAM407-13, grew optimally in media with 3–6 % NaCl and at 25–30 °C, and all could utilize n-hexadecane and n-tetradecane as the sole carbon source. The strains had a 16S rRNA gene sequence similarity of 94·2–94·3 % to Marinobacter hydrocarbonoclasticus ATCC 27132, and a similarity of 97·5–97·8 % to the closest phylogenetically related type strain, Marinobacter flavimaris DSM 16070T. DNA–DNA hybridization levels to M. flavimaris and other Marinobacter type strains were ⩽42 %, while DNA–DNA reassociation values among DG893T, DG1136 and ATAM407-13 were ⩾83 %. The DNA G+C content was 54–55 mol% and the major isoprenoid quinone was ubiquinone-9. On the basis of phenotypic, chemotaxonomic, DNA–DNA hybridization and phylogenetic analysis, it is proposed that these three strains represent a novel species, Marinobacter algicola sp. nov. The type strain is DG893T (=DSM 16394T=NCIMB 14009T).

An aerobic, Gram-negative bacterial isolate, strain DX5-10T, was isolated from coastal sediment of the East China Sea. The taxonomy of strain DX5-10T was studied by phenotypic and phylogenetic methods. Strain DX5-10T was motile, formed faint-yellowish colonies and was positive for catalase reaction and weakly positive for oxidase reaction. The nearly complete 16S rRNA gene of strain DX5-10T was obtained and sequence analysis indicated that strain DX5-10T represented an independent lineage within the Roseobacter clade of Alphaproteobacteria. Strain DX5-10T was phylogenetically related to members of the genera Roseobacter, Loktanella, Roseisalinus, Silicibacter, Antarctobacter, Sulfitobacter, Salipiger, Ruegeria and Roseivivax, and the sequence identities among them were less than 95·0 %. The predominant respiratory ubiquinone of strain DX5-10T was Q-10 and the DNA G+C content of strain DX5-10T was 63·3 mol%. Therefore, strain DX5-10T represents a novel species of a novel genus, for which the name Yangia pacifica gen. nov., sp. nov. is proposed, with the type strain DX5-10T (=CGMCC 1.3455T=JCM 12573T).

A Gram-negative, rod-shaped, non-motile, non-spore-forming, pink-pigmented bacterium, SV97T, was isolated from a wetland soil near Ny-Ålesund, Svalbard Islands, Norway (78° N). On the basis of 16S rRNA gene sequence similarity, strain SV97T was shown to belong to the Alphaproteobacteria and was highly related to a number of non-characterized Methylocystis strains with GenBank accession nos AJ458507 and AJ458502 (100 %) and AF177299, AJ458510, AJ458467, AJ458471, AJ431384, AJ458475, AJ458484, AJ458501 and AJ458466 (99 %). The most closely related type strains were Methylocystis parvus OBBPT (97·2 %) and Methylocystis echinoides IMET 10491T (97 %). The closest related recognized species within the genus Methylosinus was Methylosinus sporium NCIMB 11126T (96·0 % similarity). Chemotaxonomic and phenotypic data (C18 : 1 ω8 as the major fatty acid, non-motile, no rosette formation) supported the affiliation of strain SV97T to the genus Methylocystis. The results of DNA–DNA hybridization and physiological and biochemical tests allowed genotypic and phenotypic differentiation of strain SV97T from the two recognized Methylocystis species. Strain SV97T therefore represents a novel species, for which the name Methylocystis rosea sp. nov. is proposed, with the type strain SV97T (=DSM 17261T=ATCC BAA-1196T).

A Gram-negative bacterium, designated LA33BT, was isolated from water collected from a hypersaline lake on uninhabited Laysan Atoll in the Northwestern Hawaiian Islands. Cells of strain LA33BT are motile, straight rods that grow between 4 and 45 °C and in media containing 1–17·5 % (w/v) NaCl. The strain oxidizes carbohydrates, nucleosides, amino acids and organic acids presented as sole carbon sources and constitutive lipolytic and proteolytic enzymes are expressed. Over 75 % of the fatty acid pool is cis-11-octadecenoic acid (18 : 1ω7c). Comparative sequence analysis of the 16S rRNA gene indicates that the strain forms a new lineage in the α-2 subclass of the Proteobacteria, with the closest recognized strains being Stappia aggregata NCIMB 2208T and Roseibium denhamense JCM 10543T, with which it shares 94–95 % sequence similarity. Strain LA33BT differs phenotypically from extant Stappia and Roseibium species, however, in that it is a moderate thermophile, it requires NaCl and tolerates higher NaCl concentrations and it does not express β-galactosidase or oxidize glycerol. On the basis of genotypic data and phenotypic characteristics, we propose that strain LA33BT does not belong to the genera Stappia or Roseibium and that it represents the type species of a new genus, Nesiotobacter. Strain LA33BT (=ATCC BAA-994T=CIP 108449T) is proposed as the type strain of the type species of this genus, with the name Nesiotobacter exalbescens gen. nov., sp. nov.

Five Gram-negative, motile, spiral-shaped strains were isolated from a sulfide spring (D-412T), active sludge of wastewater (D-419T, D-420, D-424) and industrial wastewater (D-416). Comparative 16S rRNA gene sequence analysis showed that the isolates belong to the family Comamonadaceae, within the class Betaproteobacteria, but fall into a distinct cluster. On the basis of phenotypic, chemotaxonomic and phylogenetic data, a new genus, Giesbergeria gen. nov., is proposed, including five species. The type species of the genus is Giesbergeria voronezhensis sp. nov. (type strain D-419T=DSM 12825T=CIP 107340T=VKM B-2350T) and other novel members of the genus are Giesbergeria kuznetsovii sp. nov. (type strain D-412T=DSM 12827T=VKM B-2352T), Giesbergeria giesbergeri comb. nov. (basonym Aquaspirillum giesbergeri), Giesbergeria sinuosa comb. nov. (basonym Aquaspirillum sinuosum) and Giesbergeria anulus comb. nov. (basonym Aquaspirillum anulus). Using the same criteria, isolate D-416 (=DSM 12826) was identified as a strain of [Aquaspirillum] metamorphum. Strain D-416, the type strain of [A.] metamorphum and the type strain of [Aquaspirillum] psychrophilum form a distinct cluster within the family Comamonadaceae (97–97·2 % 16S rRNA gene sequence similarity) and share phenotypic and chemotaxonomic properties. Therefore, it is proposed that these strains are reclassified as members of a new genus, Simplicispira gen. nov., as Simplicispira metamorpha comb. nov. (the type species) and Simplicispira psychrophila comb. nov., respectively.

Two Gram-negative, rod-shaped, non-spore-forming bacteria (CCUG 39402T and CCUG 39797), isolated from different water sources, were investigated in a polyphasic study. The two isolates shared 100 % 16S rRNA gene sequence similarity and it was shown that they belonged to the Betaproteobacteria, most closely related to Polaromonas vacuolata (97·8 %) and Polaromonas naphthalenivorans (97·8 %). A polyamine pattern with 2-hydroxyputrescine and putrescine, as well as ubiquinone Q-8, were in agreement with characteristics of Betaproteobacteria. The presence of diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine, and major fatty acids C16 : 1 ω7c, C16 : 0 and C17 : 0 cyclo supported the affiliation of the two strains to the genus Polaromonas. The results of DNA–DNA hybridization and physiological and biochemical tests allowed genotypic and phenotypic differentiation of the two isolates from the two Polaromonas species with validly published names. They therefore represent a novel species, for which the name Polaromonas aquatica sp. nov. is proposed, with the type strain CCUG 39402T (=CIP 108776T).

Two Gram-negative, rod-shaped bacterial strains, KSL-102T and KSL-110, were isolated from an alkaline soil in Korea, and their taxonomic positions were investigated by use of a polyphasic study. The two strains grew optimally at pH 7·0–8·0 and 30 °C without NaCl. They contained Q-10 as the predominant ubiquinone. The major fatty acids were C18 : 1 ω7c and C16 : 0 on trypticase soy agar, but 11-methyl C18 : 1 ω7c was also a major component when the two strains were cultivated on LMG medium no. 221. Their DNA G+C contents were 68·4–68·7 mol%. Strains KSL-102T and KSL-110 exhibited three nucleotide differences in their 16S rRNA gene sequences and a mean DNA–DNA relatedness value of 85 %. Phylogenetic trees based on 16S rRNA gene sequences showed that the two strains fell within the evolutionary radiation encompassed by the genus Brevundimonas. Levels of 16S rRNA gene sequence similarity between the two strains and the type strains of recognized Brevundimonas species ranged from 96·3 to 98·4 %. DNA–DNA relatedness levels between the two strains and recognized Brevundimonas species were 8–21 %. On the basis of phenotypic, phylogenetic and genetic data, strains KSL-102T and KSL-110 were classified in the genus Brevundimonas as members of a novel species, for which the name Brevundimonas kwangchunensis sp. nov. is proposed. The type strain is KSL-102T (=KCTC 12380T=DSM 17033T).

Two strains of free-living diazotrophs isolated from soil from a rice paddy field were characterized by using a polyphasic approach. The novel strains, A-7T and A-4, were found to be very closely related, with 99·9 % 16S rRNA gene sequence similarity and a DNA–DNA hybridization value of 89·5 %, suggesting that they represent a single species. 16S rRNA gene sequence analyses indicated that the two strains fell within the Zoogloea lineage, with less than 96·7 % sequence similarity to other Zoogloea species. Chemotaxonomic characteristics of the novel strains, including DNA G+C content (65·1 mol%), the major quinone system (Q-8), predominant fatty acids (16 : 1ω7c and 16 : 0) and major hydroxy fatty acids (3-OH 10 : 0 and 3-OH 12 : 0), are similar to those of the genus Zoogloea. The novel strains showed positive results for floc formation which is accepted as confirmatory for species of the genus Zoogloea. However, the novel strains can be distinguished from the other species of Zoogloea by physiological characteristics. The name Zoogloea oryzae sp. nov. is therefore proposed for the novel strains with strain A-7T (=IAM 15218T=CCTCC AB 2052005T) as the type strain. Phylogenetic and chemotaxonomic analyses indicate that strain ATCC 19623, designated as a reference strain of Zoogloea ramigera, does not belong to the genus Zoogloea but to a new genus of Alphaproteobacteria. The name Crabtreella saccharophila gen. nov., sp. nov. is proposed for strain ATCC 19623T (=IAM 12669T).