Lyudmila A. Romanenko

Psychrobacter submarinus sp. nov. and Psychrobacter marincola sp. nov., psychrophilic halophiles from marine environments

Two novel psychrophilic, halophilic, Psychrobacter-like bacteria, strains KMM 225T and KMM 277T, were isolated from sea water and the internal tissues of an ascidian Polysyncraton sp. specimen, respectively, and characterized using a polyphasic approach, which included phenotypic, genotypic, chemotaxonomic and phylogenetic analyses. The novel marine isolates were Gram-negative, aerobic, coccoid, oxidase- and catalase-positive, non-pigmented, non-motile, psychrophilic and halophilic and they utilized a restricted spectrum of carbon sources. Strains KMM 225T and KMM 277T required sea water or sodium ions for growth and were tolerant of up to 12-15% (w/v) NaCl. Growth of strains KMM 225T and KMM 277T was observed at 4-35 and 7-35 degrees C, respectively. The DNA G+C contents of KMM 225T and KMM 277T were respectively 46-8 and 50.7 mol %. Comparison of almost complete 16S rDNA sequences of strains KMM 225T and KMM 277T revealed that both strains were phylogenetically most closely related to each other (99.9% sequence similarity) and slightly less related to Psychrobacter glacincola, with 97.2 and 97.8% similarity, respectively. DNA-DNA reassociation between KMM 225T and KMM 277T revealed 15% similarity, whereas similarity to other Psychrobacter species was 14-25%. Strains KMM 225T and KMM 277T differed from one another in their growth temperature, organic substrate utilization, antibiotic sensitivity and DNA G+C content. Both strains examined could be distinguished from all previously described Psychrobacter species by their physiological, genotypic and phylogenetic characteristics. On the basis of the physiological and molecular properties of the novel isolates, the names Psychrobacter submarinus sp. nov. (type strain KMM 225T = DSM 14161T) and Psychrobacter marincola sp. nov. (type strain KMM 277T = DSM 14160T) are proposed.

Structure of an acidic O-specific polysaccharide of Pseudoalteromonas haloplanktis type strain ATCC 14393 containing 2-acetamido-2-deoxy-d- and -l-galacturonic acids and 3-(N-acetyl-d-alanyl)amino-3,6-dideoxy-d-glucose

Abstract An acidic O-specific polysaccharide was obtained from the lipopolysaccharide of Pseudoalteromonashaloplanktis ATCC 14393 and found to contain d -galactose, 3-(N-acetyl- d -alanyl)amino-3,6-dideoxy- d -glucose ( d -Qui3N d AlaAc), 2,4-diacetamido-2,4,6-trideoxy- d -glucose ( d -QuiNAc4NAc), 2-acetamido-2-deoxy- d - and - l -galacturonic acids ( d - and l -GalNAcA), and O-acetyl groups. On the basis of Smith degradation and 1H and 13C NMR spectroscopic studies, including 2D COSY, TOCSY, NOESY, 1H, 13C HMQC, and HMBC experiments, the following structure of the pentasaccharide repeating unit of the polysaccharide was established: →4 )- α - l - Gal p NAcA -(1 →3 )- β - d - Qui p NAc4NAc -(1 →2 )- β - d - Qui p3 N d AlaAc -(1 →4 )- α - d - Gal p NAcA -(1 →4 )- α - d - Gal p2,6 Ac 2 -(1 → where O-acetylation of the galactose residue at each position is partial (50–70%).

Lysobacter spongiicola sp. nov., isolated from a deep-sea sponge

An aerobic, Gram-negative bacterium, strain KMM 329(T), was isolated from a deep-sea sponge specimen from the Philippine Sea and subjected to a polyphasic taxonomic investigation. Comparative 16S rRNA gene sequence analysis showed that strain KMM 329(T) clustered with the species of the genus Lysobacter. The highest level of 16S rRNA gene sequence similarity (97.0 %) was found with respect to Lysobacter concretionis KCTC 12205(T); lower values (96.4-95.2 %) were obtained with respect to the other recognized Lysobacter species. The value for DNA-DNA relatedness between strain KMM 329(T) and L. concretionis KCTC 12205(T) was 47 %. Branched fatty acids 16 : 0 iso, 15 : 0 iso, 11 : 0 iso 3-OH and 17 : 1 iso were found to be predominant. Strain KMM 329(T) had a DNA G+C content of 69.0 mol%. On the basis of the phenotypic, chemotaxonomic, DNA-DNA hybridization and phylogenetic data, strain KMM 329(T) represents a novel species of the genus Lysobacter, for which the name Lysobacter spongiicola sp. nov. is proposed. The type strain is KMM 329(T) (=NRIC 0728(T) =JCM 14760(T)).

Structure of a highly acidic O-specific polysaccharide of lipopolysaccharide of Pseudoalteromonas haloplanktis KMM 223 (44-1) containing l-iduronic acid and d-QuiNHb4NHb

An acidic O-specific polysaccharide was obtained by mild acid degradation of the lipopolysaccharide isolated by phenol-water extraction of Pseudoalteromonas haloplanktis strain KMM 223 (44-1). L-Iduronic acid (IdoA) was found to be a component of the polysaccharide and identified by NMR spectroscopy and after carboxyl-reduction followed by acid hydrolysis and acetylation, by GLC-MS as 2,3,4-tri-O-acetyl-1,6-anhydroidose. On the basis of 1H and 13C NMR spectroscopic studies, including 1D NOE, 2D NOESY, HSQC and HMBC experiments, the following structure of the branched pentasaccharide repeating unit of the polysaccharide was established: -->4)-beta-D-GlcpAI-(1-->4)-beta-D-GlcpAII-(1-->3)-beta-D-++ +QuipNHb4NHbII- (1-->2)-alpha-L-IdopA-(-->4 increases 1 alpha-D-QuipNAc4NAcI where QuiNAc4NAc and QuiNHb4NHb are 2,4-diacetamido-2,4,6-trideoxyglucose and 2,4,6-tri-deoxy-2,4- di[(S)-3-hydroxybutyramido]glucose, respectively. This is the first report of L-iduronic acid in a lipopolysaccharide and of D-QuiNHb4NHb in nature.

Halomonas halocynthiae sp. nov., isolated from the marine ascidian Halocynthia aurantium

The marine bacterium strain KMM 1376T was isolated from gill tissue of the ascidian Halocynthia aurantium, an inhabitant of the coastal waters of the Sea of Japan. Strain KMM 1376T is an aerobic, gram-negative, non-motile, encapsulated, non-pigmented, slightly halophilic bacterium that is characterized by fimbria-like structures, growth in 0.5-15% NaCl at 7-35 degrees C and absence of acid production from many carbohydrates. The DNA G+C content is 54 mol%. The main fatty acids are C16:0, C16.1omega7c and C18:1omega9c. Comparative 16S rDNA sequence analysis of strain KMM 1376T revealed 92.7-95.3% sequence similarity to members of Halomonas. Based on phenotypic and phylogenetic characteristics, it is proposed that the novel isolate be classified as Halomonas halocynthiae sp. nov., with the type strain KMM 1376T (= DSM 14573T).

Structure of an acidic polysaccharide from a marine bacterium Pseudoalteromonas distincta KMM 638 containing 5-acetamido-3,5,7,9-tetradeoxy-7- formamido-L-glycero-L-manno-nonulosonic acid

An acidic polysaccharide was obtained from the lipopolysaccharide of Pseudoalteromonas distincta strain KMM 638, isolated from a marine sponge, and found to contain D-GlcA, D-GalNAc, 2-acetamido-2,6-dideoxy-D-glucose (D-QuiNAc) and two unusual acidic amino sugars: 2-acetamido-2-deoxy-D-galacturonic acid (D-GalNAcA) and 5-acetamido-3,5,7,9-tetradeoxy-7-formamido-L-glycero-L-manno-nonulosonic acid (Pse5Ac7Fo, a derivative of pseudaminic acid). Oligosaccharides were derived from the polysaccharide by partial acid hydrolysis and mild alkaline degradation and characterised by electrospray ionisation (ESI) MS and 1H and 13C NMR spectroscopy. Based on these data and NMR spectroscopic studies of the initial and O-deacetylated polysaccharides, including quaternary carbon detection, 2D COSY, TOCSY, ROESY, H-detected 1H,13C HMQC and HMBC experiments, the following structure of the branched pentasaccharide repeating unit was established: [structure: see text].

Winogradskyella arenosi sp. nov., a member of the family Flavobacteriaceae isolated from marine sediments from the Sea of Japan.

An aerobic, Gram-negative, yellow-pigmented, gliding bacterial strain, designated R60(T), was isolated from a marine sediment sample obtained from the Sea of Japan and was characterized by using a polyphasic approach. 16S rRNA gene sequence analysis showed that strain R60(T) belonged to the genus Winogradskyella, sharing <97 % sequence similarity with the type strains of recognized Winogradskyella species. The main fatty acids of strain R60(T) were iso-C(15 : 0) 3-OH, iso-C(16 : 0) 3-OH, iso-C(17 : 0) 3-OH, anteiso-C(17 : 0) 3-OH and iso-C(15 : 0), consistent with its assignment to the genus Winogradskyella. On the basis of phenotypic distinctiveness and phylogenetic divergence, strain R60(T) is considered to represent a novel species of the genus Winogradskyella, for which the name Winogradskyella arenosi sp. nov. is proposed. The type strain is R60(T) (=KMM 3968(T) =NRIC 0748(T)= JCM 15527(T)).

Occurrence and antagonistic potential of Stenotrophomonas strains isolated from deep-sea invertebrates

Stenotrophomonas maltophilia is known to be of significance as opportunistic pathogen as well as a source of biocontrol and bioremediation activities. S. maltophilia strains have been isolated from rhizospheres, soil, clinical material, aquatic habitats, but little is known about Stenotrophomonas strains recovered from marine environments. During a survey of the biodiversity of Pseudomonas-like bacteria associated with deep-sea invertebrates six Stenotrophomonas strains were isolated from sponge, sea urchin, and ophiura specimens collected from differing Pacific areas, including the Philippine Sea, the Fiji Sea and the Bering Sea. 16S rRNA gene sequence analysis confirmed an assignment of marine isolates to the genus Stenotrophomonas as it placed four strains into the S. maltophilia CIP 60.77T cluster and two related to the S. rhizophila DSM 14405T. Together with a number of common characteristics typical of S. maltophilia and S. rhizophila marine isolates exhibited differences in pigmentation, a NaCl tolerance, a range of temperatures, which supported their growth, substrate utilization pattern, and antibiotics resistance. Strains displayed hemolytic and remarkable inhibitory activity against a number of fungal cultures and Gram-positive microorganisms, but very weak or none against Candida albicans. This is the first report on isolation, taxonomic characterization and antimicrobial activity of Stenotrophomonas strains isolated from deep-sea invertebrates.

Luteimonas vadosa sp. nov., isolated from seashore sediment.

An aerobic, Gram-stain-negative, yellow-pigmented, non-motile, rod-shaped bacterium designated strain KMM 9005(T) was isolated from a sediment sample collected from the Sea of Japan seashore. Comparative 16S rRNA gene sequence analysis showed that strain KMM 9005(T) belonged to the genus Luteimonas and was most closely related to Luteimonas cucumeris KCTC 23627(T) (96.5 % sequence similarity) and Luteimonas aquatica LMG 24212(T) (96.1 % sequence similarity). Strain KMM 9005(T) was characterized by the presence of thin fimbriae, the major ubiquinone Q-8, by the predominance of iso-C17 : 1 followed by iso-C16 : 0, iso-C15 : 0 and iso-C17 : 0 in its fatty acid profile, weak hydrolytic capacity and the inability to assimilate most organic substrates. Based on these distinctive phenotypic characteristics and phylogenetic analysis, strain KMM 9005(T) represents a novel species of the genus Luteimonas, for which the name Luteimonas vadosa sp. nov. is proposed. The type strain is KMM 9005(T) ( = NRIC 0881(T) = JCM 18392(T)).

Description of Cobetia amphilecti sp. nov., Cobetia litoralis sp. nov. and Cobetia pacifica sp. nov., classification of Halomonas halodurans as a later heterotypic synonym of Cobetia marina and emended descriptions of the genus Cobetia and Cobetia marina

A group of five Gram-negative, aerobic, halotolerant, non-pigmented bacteria isolated from shallow sediment samples and invertebrate specimens collected from the Gulf of Alaska and the Sea of Japan was subjected to taxonomic study. On the basis of 16S rRNA gene sequence analysis, the novel isolates were affiliated to the genus Cobetia, sharing the highest sequence similarity of 99.3-99.9 % with Cobetia marina DSM 4741(T). DNA-DNA hybridization experiments between and among the novel strains and C. marina DSM 4741(T) and Cobetia crustatorum JCM 15644(T) revealed that the five strains represent three separate genospecies, which could be differentiated in their morphological, physiological and biochemical characteristics. Halomonas halodurans NBRC 15607(T) was included in this study as it has recently been reported to exhibit high 16S rRNA gene sequence similarity to C. marina DSM 4741(T), and it showed a high DNA relatedness value of 96 % with C. marina DSM 4741(T), indicating that they belong to the same species. On the basis of phylogenetic analysis, DNA-DNA hybridization and phenotypic characterization, three novel species are proposed, named Cobetia amphilecti sp. nov. (type strain KMM 1561(T) = NRIC 0815(T) = CCUG 49560(T)), Cobetia litoralis sp. nov. (type strain KMM 3880(T) =NRIC 0814(T) =CCUG 49563(T)) and Cobetia pacifica sp. nov. (type strain KMM 3879(T) = NRIC 0813(T) = CCUG 49562(T)). It is also proposed that Halomonas halodurans is a later heterotypic synonym of Cobetia marina, and emended descriptions of the genus Cobetia and the species Cobetia marina are provided.