Anatolii M. Lysenko

Arenibacter gen. nov., new genus of the family Flavobacteriaceae and description of a new species, Arenibacter latericius sp. nov

Five dark-orange-pigmented, Gram-negative, rod-shaped, non-motile, aerobic bacterial strains were isolated from sandy sediment samples collected in the South China Sea in the Indian Ocean, from a holothurian, Apostichopus japonicus, in the Sea of Japan and from a brown alga, Chorda filum, from the Sea of Okhotsk in the Pacific Ocean. Phenotypic data were collected, demonstrating that the bacteria are chemo-organotrophic and require seawater-based media for growth. Polar lipids were analysed and 27% of the total extract comprised phosphatidylethanolamine as the major component. The predominant cellular fatty acids were branched-chain saturated and unsaturated [i-C15:0, i-C15:1, a-C15:0, C15:0, C16:1(n-7)]. The DNA base composition was 37.5-38.2 mol % G+C. The level of DNA homology of the five isolates was 83-94%, indicating that these isolates belong to the same species. A 16S rDNA sequence of the type strain KMM 426T was determined and phylogenetic analysis, based on neighbour-joining and Fitch-Margoliash methods, revealed that the type strain formed a distinct phyletic line in a clade corresponding to the family Flavobacteriaceae and represented a new genus. From the results of this polyphasic taxonomic analysis, it is proposed that the bacterial strains be classified in a new genus, Arenibacter gen. nov., and species, Arenibacter latericius sp. nov. The type strain is KMM 426T (VKM B 2137DT = LMG 19694T = CIP 106861T).

Pseudoalteromonas issachenkonii sp. nov., a bacterium that degrades the thallus of the brown alga Fucus evanescens.

Eleven non-pigmented strains of gram-negative, aerobic, marine bacteria with polar flagella were isolated from the thallus of the brown alga Fucus evanescens collected in the Kraternaya Bight of the Kurile Islands in the Pacific Ocean. These organisms were conspecific and exhibited high levels of genetic relatedness (up to 91%). The G+C contents of the DNAs of these strains were 42.9-43.3 mol%. These halophilic bacteria had bacteriolytic, proteolytic and haemolytic activities and degraded algal polysaccharides, synthesizing a number of glycoside hydrolases (fucoidanases, laminaranases, alginases, agarases, pullulanases, beta-glucosidases, beta-galactosidases, beta-N-acetylglucosaminidases and beta-xylosidases). By 16S rDNA analysis, the bacteria were shown to belong to the genus Pseudoalteromonas, a member of the gamma-subclass of the Proteobacteria. DNA from the strains isolated from the brown alga showed 27-54% genetic relatedness with respect to DNAs of other type strains of the genus Pseudoalteromonas. The phenotypic characteristics, together with the genetic evidence, indicate that this group of epiphytic bacteria represents a distinct species, Pseudoalteromonas issachenkonii sp. nov., for which the type strain is KMM 3549T (= LMG 19697T = CIP 106858T).

Pseudoalteromonas maricaloris sp. nov., isolated from an Australian sponge, and reclassification of [Pseudoalteromonas aurantia] NCIMB 2033 as Pseudoalteromonas flavipulchra sp. nov.

A marine, gram-negative, aerobic bacterium that produced cytotoxic, lemon-yellow, chromopeptide pigments that inhibited the development of sea urchin eggs has been isolated from the Australian sponge Fascaplysinopsis reticulata Hentschel. The cells of the organism were rod-shaped with a single polar flagellum and they required NaCl for growth (0.5-10%) with optimum growth at 1-3% NaCl. The temperature for growth was 10-37 degrees C, with optimum growth at 25-30 degrees C. Growth occurred at pH values from 6.0 to 10.0, with optimum growth at pH 6.0-8.0. Major phospholipids were phosphatidylethanolamine, phosphatidylglycerol and lyso-phosphatidylethanolamine. Of 26 fatty acids with 11-19 carbon atoms that were detected, 16:1omega7, 16:0, 17:1omega8 and 18:1omega7 were predominant. The DNA G+C content was 38.9 mol%. All of these phenotypic and chemotaxonomic characters place the organism in the genus Pseudoalteromonas (Gauthier et al, 1995). These data are consistent with the phylogenetic analyses that confirmed that strain KMM 636T is a member of the Pseudoalteromonas cluster in the gamma-subclass of the Proteobacteria. DNA-DNA hybridization experiments revealed that the levels of relatedness between the DNA of the strain studied and DNAs of type strains of the species that clustered together (on the basis of 16S rDNA sequences) and [Pseudoalteromonas aurantia] NCIMB 2033 ranged from 19 to 35%, and that the DNA-DNA homology between [P. aurantia] NCIMB 2033 and other phylogenetically and/or phenotypically similar type strains ranged from 32 to 52%. According to the polyphasic evidence presented in this study, it is proposed that strain KMM 636T (= LMG 19692T = CIP 106859T) be classified as Pseudoalteromonas maricaloris sp. nov. and [P. aurantia] NCIMB 2033 be reclassified as Pseudoalteromonas flavipulchra NCIMB 2033T (= KMM 3630T = LMG 20361T) sp. nov.

Pseudoalteromonas ruthenica sp. nov., isolated from marine invertebrates

On the basis of phenotypic and genotypic characteristics and analysis of 165 rRNA sequences, a novel species belonging to the genus Pseudoalteromonas is described. Two pale-orange-pigmented strains, KMM 300T and KMM 290, isolated respectively from a mussel, Crenomytilus grayanus, and a scallop, Patinopecten yessoensis, are marine, gram-negative, aerobic, rod-shaped bacteria that produce a number of antimicrobial compounds. The strains are able to degrade gelatin, elastin, starch, DNA and Tween 80. Chitin and agar are not degraded. The isolates from marine invertebrates grew at NaCl concentrations of 1-9% and a temperature range of 10-35 degrees C and did not utilize most of the wide range of carbohydrates tested, with the exception of D-glucose, cellobiose and sucrose. The DNA G+C content was 48.4-48.9 mol%. The level of DNA homology of the two strains was 98%. DNA from the strains isolated from marine invertebrates showed 5-15% genetic relatedness to the DNA of other type strains of the genus Pseudoalteromonas. 16S rRNA analysis indicated a clear affiliation of the novel bacteria to other species of the genus. The strains are assigned to a novel species, Pseudomonas ruthenica sp. nov., with the type strain KMM 300T (= LMG 19699T = CIP 106857T).

Pseudomonas extremorientalis sp. nov., isolated from a drinking water reservoir.

On the basis of phenotypic and genotypic characteristics and 16S rDNA sequence analysis, a novel species belonging to the genus Pseudomonas sensu stricto was identified. The saprophytic, fluorescent bacterium, designated KMM 3447(T), was isolated from a drinking water reservoir near Vladivostok City, Russia. The novel organism was a Gram-negative, aerobic, rod-shaped bacterium that produced a cyclic depsipeptide with surface-active properties. It degraded casein, but did not degrade gelatin, starch, agar or Tween 80. The bacterium was also haemolytic. Growth of the novel bacterium occurred between 4 and 35 degrees C. The predominant cellular fatty acids of the novel pseudomonad were C16:0, C16:1(n-7), C18:1(n-7) and C17.0 cyclo; branched fatty acids were only found in trace amounts. The G+C content of the novel bacterium was 61.0 mol%. 16S rDNA sequence analysis indicated that the novel bacterium had a clear affiliation with Pseudomonas fluorescens and species closely related to this recognized pseudomonad. DNA-DNA hybridization experiments showed that the novel bacterium bound at low levels (27-53%) with the DNA of the type strains of its nearest phylogenetic relatives, namely Pseudomonas tolaasii, Pseudomonas veronii, Pseudomonas orientalis and Pseudomonas rhodesiae, indicating that the novel bacterium represented a novel species within the genus Pseudomonas, for which the name Pseudomonas extremorientalis is proposed; the type strain is KMM 3447(T) (= LMG 19695(T)).

Retrieval of the species Alteromonas tetraodonis Simidu et al. 1990 as Pseudoalteromonas tetraodonis comb. nov. and emendation of description.

A polyphasic taxonomy study was undertaken of three strains of Pseudoalteromonas haloplanktis subsp. tetraodonis (Simidu et al. 1990) Gauthier et al. 1995. DNA was prepared from each of the strains and genomic relatedness was measured by DNA-DNA hybridization. Strains KMM 458T and IAM 14160T shared 99% genetic relatedness, but were only 48-49% related to the type strain of Pseudoalteromonas haloplanktis subsp. haloplanktis, IAM 12915T. The third strain, P. haloplanktis subsp. tetraodonis A-M, showed 83% genetic similarity with P. haloplanktis subsp. haloplanktis IAM 12915T and 32% with KMM 458T. From these results, it is concluded that strains KMM 458T and IAM 14160T comprise a separate species, originally described as Alteromonas tetraodonis, whereas strain A-M belongs to the species Pseudoalteromonas haloplanktis. Based on phenotypic and chemotaxonomic data, genomic fingerprint patterns, DNA-DNA hybridization data and phylogenetic analysis of 16S rRNA, it is proposed that the species Alteromonas tetraodonis be retrieved and recognized as Pseudoalteromonas tetraodonis comb. nov. (type strain IAM 14160T = KMM 458T).

Pseudoalteromonas translucida sp. nov. and Pseudoalteromonas paragorgicola sp. nov., and emended description of the genus.

On the basis of phenotypic and genotypic characteristics and analysis of 16S rRNA sequences, two novel species belonging to the genus Pseudoalteromonas are described. A pale-orange-pigmented strain, KMM 3548T, isolated from a sponge and a non-pigmented strain, KMM 520T, isolated from sea water are marine, gram-negative, aerobic, rod-shaped organisms. One of the strains, KMM 520T, had bipolar flagella. Both strains had the ability to degrade gelatin, DNA and Tween 80 but not chitin or agar. Strain KMM 520T decomposed elastin and grew at NaCl concentrations of 1-8%, while strain KMM 3548T grew at 1-6% NaCl. The temperature range for both strains was 4-30 degrees C. The DNA G+C contents were 46.3 (KMM 520T) and 41.1 mol% (KMM 3548T). The level of DNA relatedness between the two strains was 20%. DNA from strain KMM 520T showed 8-34% genetic relatedness and that of KMM 3548T showed 17-53% relatedness to the DNA of other type strains of the genus Pseudoalteromonas. 16S rRNA analysis indicated a clear affiliation of these novel bacteria with the genus Pseudoalteromonas. The type strains of the novel species are Pseudoalteromonas translucida sp. nov. KMM 520T (= LMG 19696T = ATCC BAA-3157T) and Pseudoalteromonas paragorgicola sp. nov. KMM 3548T (= LMG 19694T = ATCC BAA-322T).