Aerobic degradation of cis-dichloroethene by the marine bacterium Marinobacter salsuginis strain 5N-3.

An aerobic bacterium, designated strain 5N-3 (NBRC 113055), that degrades cis-dichloroethene (cDCE) was isolated from a sea sediment in Japan. Strain 5N-3 was able to degrade a certain amount of cDCE in the presence of pyruvate without the action of inducers. In the presence of inducers, such as phenol and benzene, the strain completely removed cDCE. By the application of 16S ribosomal RNA (16S rRNA) gene sequencing and average nucleotide identity analyses, the strain 5N-3 was identified as Marinobacter salsuginis. On the other hand, identified species of Marinobacter are not known to degrade cDCE at all. A draft genome sequence analysis of the strain 5N-3 suggested that the dmp-homologous operon (operon for phenol degradation) may be contributing to the aerobic degradation of cDCE. This is the first report on an aerobic marine bacterium that has been found to degrade cDCE.

Rhizobacter gummiphilus sp. nov., a rubber-degrading bacterium isolated from the soil of a botanical garden in Japan.

A rubber-degrading bacterium, the strain NS21, that was isolated from a soil sample in a botanical garden in Japan (Imai et al., 2011) was examined by phenotypic, phylogenetic and chemotaxonomic approaches to determine its taxonomic position. The strain NS21 was motile, possessing a single polar flagellum and a facultatively anaerobic straight rod. Analysis of the 16S rRNA and gyrB gene sequences of NS21 revealed a close relationship to the genus Rhizobacter. The predominant quinone type was Q-8. The G+C content of the NS21 genomic DNA was 70.8 mol%. The major fatty acids were C16:0, C17:0 cyclo, C18:1ω7c and C16:1ω7c and/or iso-C15:0 2-OH. C12:0 2-OH was present. The DNA-DNA hybridization experiments indicated that the DNA relatedness values of the strain NS21 to R. dauci H6(T) and R. fulvus Gsoil 322(T) were lower than 24%. The phenotypic characteristics showed obvious dissimilarities when compared to closely related species. On the basis of these taxonomic properties, a novel species is proposed as Rhizobacter gummiphilus sp. nov., with the strain NS21(T) (NBRC 109400(T), BCC 58006(T)) as the type strain. The emended description of the genus Rhizobacter was also presented.

Oleibacter marinus gen. nov., sp. nov., a bacterium that degrades petroleum aliphatic hydrocarbons in a tropical marine environment.

Three Gram-negative, motile, mesophilic, aerobic, rod-shaped bacterial strains, designated 2O1(T), 1O14 and 1O18, were isolated from Indonesian seawater after enrichment with crude oil and a continuous supply of supplemented seawater. The strains exhibited high n-alkane-degrading activity, which indicated that the strains were important degraders of petroleum aliphatic hydrocarbons in tropical marine environments. Phylogenetic analyses based on 16S rRNA gene sequences of members of the Gammaproteobacteria showed that the isolates formed a coherent and distinct cluster in a stable lineage containing Oceanobacter kriegii IFO 15467(T) (96.4-96.5 % 16S rRNA gene sequence similarity) and Thalassolituus oleivorans MIL-1(T). DNA G +C content was 53.0-53.1 mol%. The major fatty acids were C(16 : 0), C(16 : 1)ω7 and C(18 : 1)ω9 and the hydroxy fatty acids were C(12 : 0) 3-OH and C(10 : 0) 3-OH. The polar lipids were phosphatidylglycerol, a ninhydrin-positive phospholipid(s) and glycolipids. The major quinone was Q-9 (97-99 %), which distinguished the isolates from Oceanobacter kriegii NBRC 15467(T) (Q-8; 91 %). On the basis of phenotypic, genotypic and chemotaxonomic data, including DNA-DNA hybridization, the isolates represent a novel genus and species, for which the name Oleibacter marinus gen. nov., sp. nov. is proposed. The type strain of Oleibacter marinus is 2O1(T) (=NBRC 105760(T) =BTCC B-675(T)).

Euzebya tangerina gen. nov., sp. nov., a deeply branching marine actinobacterium isolated from the sea cucumber Holothuria edulis, and proposal of Euzebyaceae fam. nov., Euzebyales ord. nov. and Nitriliruptoridae subclassis nov.

A tangerine-coloured, Gram-positive actinobacterial strain, designated F10(T), was isolated from the abdominal epidermis of a sea cucumber, Holothuria edulis, collected in seawater off the coast of Japan. A 16S rRNA gene sequence analysis indicated that strain F10(T) was a member of the class Actinobacteria and was most closely related to Nitriliruptor alkaliphilus ANL-iso2(T) (87.4 % sequence similarity). Phylogenetic analyses showed that strain F10(T) represented a novel, deep-rooted, and distinct phylogenetic lineage within the class Actinobacteria and clustered with N. alkaliphilus and uncultured bacteria. The organism had meso-diaminopimelic acid as the diagnostic diamino acid in the cell-wall peptidoglycan, and rhamnose and galactose as the diagnostic cell-wall sugars. Strain F10(T) contained C16 :0ω7c, C16:0 and C17:1ω8c as the major cellular fatty acids. The predominant isoprenoid quinone was MK-9 (H4).The G+C content of the DNA was 68.3 mol%. Based on data from the current polyphasic study, it is proposed that the new marine isolate be placed in a novel genus and be considered a novel species designated Euzebya tangerina gen. nov., sp. nov. within the new family, order and subclass Euzebyaceae fam. nov., Euzebyales ord. nov. and Nitriliruptoridae subclassis nov. in the class Actinobacteria. The type strain of Euzebya tangerina is F10(T) (=NBRC 105439(T) =KCTC 19736(T)).

Phycisphaera mikurensis gen. nov., sp. nov., isolated from a marine alga, and proposal of Phycisphaeraceae fam. nov., Phycisphaerales ord. nov. and Phycisphaerae classis nov. in the phylum Planctomycetes.

Three strains, FYK2301M01(T), FYK2301M18 and FYK2301M52, all being Gram-negative, spherical, motile and facultatively anaerobic, were isolated from a marine alga (Porphyra sp.) collected on Mikura Island, Japan. Colonies of the strains were circular and pink-pigmented on Marine Agar 2216 (Difco) at 25 degrees C. Cells of the strains reproduced by binary fission. The G+C content of the DNA was 73 mol%. The major isoprenoid quinone was MK-6. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the strains are the members of the WPS-1 group (Nogales et al., 2001) comprising no validly described taxa within the phylum Planctomycetes. The highest similarity value of the 16S rRNA gene sequences of the strains to those in the established bacterial taxa was only 78.7% to Planctomyces brasiliensis DSM 5305(T). From the taxonomic data obtained in this study, it is proposed that the new marine isolates be placed in a novel genus and species named Phycisphaera mikurensis gen. nov., sp. nov. within a new family, order and class Phycisphaeraceae fam. nov., Phycisphaerales ord. nov. and Phycisphaerae classis nov. in the phylum Planctomycetes. The type strain of Phycisphaera mikurensis is FYK2301M01(T) (= NBRC 102666(T) = KCTC 22515(T)).

Iamia majanohamensis gen. nov., sp. nov., an actinobacterium isolated from sea cucumber Holothuria edulis, and proposal of Iamiaceae fam. nov.

A novel, Gram-positive bacterial strain, F12(T), was isolated from the abdominal epidermis of a sea cucumber, Holothuria edulis, collected from seawater off the coast of Japan. According to 16S rRNA gene sequence analysis, this strain represents a novel, deep-rooting lineage within the class Actinobacteria and clusters with uncultured bacteria and Acidimicrobium ferrooxidans. Compared to species with validly published names, the highest 16S rRNA gene sequence similarity (89.8 %) was to Acidimicrobium ferrooxidans DSM 10331(T). Phylogenetic analyses showed that strain F12(T) represents a distinct phylogenetic lineage related closely to the genus Acidimicrobium. Strain F12(T) contained MK-9(H(6)) as the major menaquinone, whilst 17 : 0, 17 : 1omega8c, 15 : 0 and 16 : 0 were the major cellular fatty acids. The cell-wall peptidoglycan of strain F12(T) was composed of meso-diaminopimelic acid as the diagnostic diamino acid, alanine and glutamic acid (1 : 2 : 1). The cell-wall sugars detected were rhamnose, mannose, arabinose, galactose and xylose. The G+C content of the DNA was 74.4 mol%. From the taxonomic data obtained in this study, the name Iamia majanohamensis gen. nov., sp. nov. is proposed for the isolate, with type strain F12(T) (=NBRC 102561(T)=DSM 19957(T)). The name Iamiaceae fam. nov. is also proposed for the distinct phyletic line represented by the genus Iamia.

Acanthopleuribacter pedis gen. nov., sp. nov., a marine bacterium isolated from a chiton, and description of Acanthopleuribacteraceae fam. nov., Acanthopleuribacterales ord. nov., Holophagaceae fam. nov., Holophagales ord. nov. and Holophagae classis nov. in the phylum 'Acidobacteria'.

Strain FYK2218(T) was isolated from a specimen of the chiton Acanthopleura japonica, which had been collected from a beach on the Boso peninsula in Japan. Phylogenetic analyses based on 16S rRNA gene sequences revealed that the strain belonged to the phylum 'Acidobacteria'. The most closely related type strains to strain FYK2218(T) were Holophaga foetida TMBS4(T) (83.6 % 16S rRNA gene sequence similarity) and Geothrix fermentans H-5(T) (83.6 %) in subdivision 8 of the 'Acidobacteria'. Cells of FYK2218(T) were motile, rod-shaped, Gram-negative, mesophilic and strictly aerobic. The G+C content of the strain was 56.7 mol%. The strain had isoprenoid quinones MK-6 and MK-7 as major components. Major fatty acids of the strain were iso-C(15 : 0), iso-C(17 : 0), C(16 : 0) and C(20 : 5)omega3c (cis-5,8,11,14,17-eicosapentaenoic acid). From the taxonomic data obtained in this study, it is proposed that the new marine isolate be placed into a novel genus and species named Acanthopleuribacter pedis gen. nov., sp. nov. within the new family, order and class Acanthopleuribacteraceae fam. nov., Acanthopleuribacterales ord. nov. and Holophagae classis nov. The family Holophagaceae fam. nov. is also described. The type strain of Acanthopleuribacter pedis is FYK2218(T) (=NBRC 101209(T) =KCTC 12899(T)).

Sneathiella glossodoripedis sp. nov., a marine alphaproteobacterium isolated from the nudibranch Glossodoris cincta, and proposal of Sneathiellales ord. nov. and Sneathiellaceae fam. nov.

A novel marine bacterium, designated strain MKT133(T), was isolated from the foot epidermis of a nudibranch, Glossodoris cincta (Mollusca), collected in seawater off the coast of Japan at a depth of 4 m. This bacterium was Gram-negative, motile, mesophilic and strictly aerobic, with small rod-shaped cells. Colonies of the strain after 4-5 days incubation on marine agar 2216 at 30 degrees C were less than 1 mm in diameter. The strain required salt for growth and contained Q-10 as the predominant respiratory quinone, C(18 : 1) omega 7c, C(16 : 0) and C(17 : 1) as major cellular fatty acids and C(14 : 0) 3-OH as a hydroxy fatty acid. 16S rRNA gene sequence analysis showed that the isolate had highest similarity to Sneathiella chinensis, with 97.2 % sequence similarity to the type strain. Our phylogenetic analysis also revealed that this clade represents a distinct lineage and forms a deep branch with less than 90 % 16S rRNA gene sequence similarity to the members of the eight known orders within the Alphaproteobacteria. Sufficient differences exist to distinguish this strain from Sneathiella chinensis. The name Sneathiella glossodoripedis sp. nov. is proposed, with the type strain MKT133(T) (=IAM 15419(T) =KCTC 12842(T)). The novel order Sneathiellales ord. nov. and family Sneathiellaceae fam. nov. are proposed for the distinct phyletic line represented by the genus Sneathiella.

Emended descriptions of the genus Lewinella and of Lewinella cohaerens, Lewinella nigricans and Lewinella persica, and description of Lewinella lutea sp. nov. and Lewinella marina sp. nov.

Two strains, MKG-38(T) and FYK2402M69(T), were isolated from a marine sediment sample and a sea snail, respectively, both collected on the Pacific coast of Japan. Phylogeny of these new isolates based on 16S rRNA gene sequences indicated that they are members of the genus Lewinella. Morphological, physiological and biochemical properties of these two isolates, together with the type strains of the three previously described species of the genus Lewinella, were characterized. The new isolates were Gram-negative, aerobic, rod-shaped, chemo-organotrophic and able to degrade starch and CM-cellulose. A comparative polyphasic study showed that these two isolates represent two novel species of the genus Lewinella, for which the names Lewinella marina sp. nov. (type strain, MKG-38(T)=NBRC 102633(T)=NCIMB 14312(T)) and Lewinella lutea sp. nov. (type strain, FYK2402M69(T)=NBRC 102634(T)=NCIMB 14313(T)) are proposed. Emended descriptions of the genus Lewinella (Sly et al. 1998) and of Lewinella cohaerens, Lewinella nigricans and Lewinella persica are also proposed.

Pseudovibrio ascidiaceicola sp. nov., isolated from ascidians (sea squirts).

Two bacterial strains, F423T and F10102, were isolated from two ascidians, Polycitor proliferus and Botryllidae sp., respectively, which were collected from a beach on the Boso peninsula in Japan. Cells of both isolates were motile, rod-shaped and formed star-shaped aggregates in the early stage of exponential growth, but were coccoid in stationary growth phase. The results of 16S rRNA gene sequence analysis, fatty acid analysis, DNA-DNA hybridization experiments and physiological and biochemical tests indicated that the two strains were members of a novel species of the genus Pseudovibrio for which the name Pseudovibrio ascidiaceicola sp. nov. is proposed. The type strain is F423T (=NBRC 100514T=IAM 15084T=DSM 16392T=KCTC 12308T).