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A bacterial strain, designated KMM 6244(T), was isolated from the sea urchin Strongylocentrotus intermedius and subjected to a polyphasic taxonomic investigation. The bacterium was found to be heterotrophic, aerobic, non-motile and spore-forming. Comparative phylogenetic analysis based on 16S rRNA gene sequencing placed the marine isolate in the genus Bacillus. The nearest neighbor of strain KMM 6244(T) was Bacillus decolorationis LMG 19507(T) with a 16S rRNA gene sequence similarity of 98.0%. Sequence similarities with the other recognized Bacillus species were less than 96.0%. The results of the DNA-DNA hybridization experiments revealed a low relatedness (37%) of the novel isolate with the type strain of B. decolorationis LMG 19507(T). Strain KMM 6244(T) grew at 4-45°C and with 0-12% NaCl. It produced catalase and oxidase and hydrolyzed aesculin, casein, gelatin and DNA. The predominant fatty acids were anteiso-C(15:0), iso-C(15:0), anteiso-C(17:0), C(15:0), iso-C(16:0) and iso-C(14:0). The DNA G + C content was 39.4 mol%. A combination of phylogenetic, genotypic and phenotypic data clearly indicated that strain KMM 6244(T) represents a novel species in the genus Bacillus, for which the name Bacillus berkeleyi sp. nov. is proposed. The type strain is KMM 6244(T) (KCTC 12718(T) = LMG 26357(T)).

A Gram-stain-negative, aerobic, yellow-pigmented, heterotrophic, sulfur-oxidizing, non-motile, rod-shaped bacterium, designated KMM 3898(T), was isolated from an internal tissue of the sand snail Umbonium costatum, collected from the shallow sediments of the Sea of Japan. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain KMM 3898(T) formed a distinct phylogenetic lineage within the class Gammaproteobacteria and was most closely related to Leucothrix mucor DSM 2157(T) (89.2 % 16S rRNA gene sequence similarity) and members of the genus Thiothrix (86.7-88.5 %). Chemotaxonomically, strain KMM 3898(T) contained the isoprenoid quinone Q-8, the polar lipids phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and an unknown phospholipid and the fatty acids C(18 : 1)ω7c, C(16 : 1)ω7c and C(16 : 0) as predominant components (>10 %). The DNA G+C content of strain KMM 3898(T) was 43.4 mol%. On the basis of phenotypic features and phylogenetic analysis, strain KMM 3898(T) represents a novel genus and species, for which the name Cocleimonas flava gen. nov., sp. nov. is proposed. The type strain is KMM 3898(T) (=NRIC 0757(T) =JCM 16494(T)).

Two Gram-negative, strictly aerobic, non-motile bacteria designated strains KMM 3851(T) and KMM 3842(T) were respectively isolated from a marine snail specimen (Umbonium costatum) and from surrounding sediments collected simultaneously from the shore of the Sea of Japan. Phylogenetic analysis based on 16S rRNA gene sequences showed that strains KMM 3851(T) and KMM 3842(T) were affiliated with the Roseobacter lineage of the class Alphaproteobacteria as a separate phylogenetic line adjacent to the members of the genus Thalassobacter. These novel isolates shared 98.5 % 16S rRNA gene sequence similarity and 15 % DNA-DNA relatedness. The major isoprenoid quinone of both strains was Q-10 and polar lipids consisted of phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol and aminophospholipids. In addition, strain KMM 3851(T) contained two unknown lipids, whereas strain KMM 3842(T) contained diphosphatidylglycerol. Fatty acid analysis revealed C(18 : 1)ω7c and C(16 : 0) as major components and small amounts of C(18 : 2). The DNA G+C contents were 60.4 mol% (KMM 3851(T)) and 58.5 mol% (KMM 3842(T)). Based on distinctive phenotypic characteristics, DNA-DNA hybridization data and phylogenetic distance, strains KMM 3851(T) and KMM 3842(T) should be classified as representatives of two novel species in a new genus, Litoreibacter gen. nov., with the type species Litoreibacter albidus sp. nov. (type strain KMM 3851(T) =NRIC 0773(T) =JCM 16493(T)) and a second species Litoreibacter janthinus sp. nov. (type strain KMM 3842(T) =NRIC 0772(T) =JCM 16492(T)).

Three Gram-negative, aerobic, halophilic, non-motile, yellowish-pigmented bacteria, designated KMM 3896, KMM 3897 and KMM 3899(T), were isolated from coastal seawater and sediment samples of the Sea of Japan, Russia. The isolates were shown to belong to the same species on the basis of 16S rRNA gene sequence similarities (99.6-99.9 %) and DNA-DNA relatedness (73-98 %). Phylogenetic analysis of 16S rRNA gene sequences demonstrated that the isolates formed a subcluster within the genus Kangiella in the class Gammaproteobacteria. 16S rRNA gene sequence similarities between strain KMM 3899(T) and its closest phylogenetic neighbours, Kangiella koreensis SW-125(T) and Kangiella aquimarina SW-154(T), were 96.6 and 95.5 %, respectively. On the basis of phenotypic differences and phylogenetic distances, it is proposed that strains KMM 3896, KMM 3897 and KMM 3899(T) are assigned to a novel species of the genus Kangiella, Kangiella japonica sp. nov. The type strain is KMM 3899(T) (=NRIC 0764(T) =JCM 16211(T)).

A Gram-negative, aerobic, yellow-pigmented, rod-shaped, non-motile bacterium, strain KMM 3895T, was isolated from a marine sandy sample collected offshore from the Sea of Japan. Phylogenetic analysis based on the 16S rRNA gene sequence placed strain KMM 3895T in the class Gammaproteobacteria, forming a separate branch, sharing 89.5 % sequence similarity with Nitrincola lacisaponensis 4CAT and 88-87% similarity with the other members of the cluster, including members of Kangiella, Spongiibacter, Alcanivorax and Microbulbifer. The major isoprenoid quinone was Q-8. Polar lipids consisted of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, aminophospholipid and an unknown phospholipid. Fatty acid analysis revealed C16:1omega7, iso-C16:0, iso-C18:0 and C18:1omega7 as dominant components. The DNA G+C content was determined to be 48.1 mol%. Based on its unique phenotypic characteristics and phylogenetic distance, the marine coastal sediment isolate KMM 3895T should be classified as a representative of a novel genus and species, for which the name Arenicella xantha gen. nov., sp. nov. is proposed. The type strain of Arenicella xantha is KMM 3895T (=NRIC 0759T =JCM 16153T).

A Gram-negative, aerobic, greyish-yellowish-pigmented, stenohaline, rod-shaped, non-motile bacterium, strain KMM 3900(T), was isolated from a coastal seawater sample collected from the Sea of Japan. Based on phylogenetic analysis, strain KMM 3900(T) was positioned within the Gammaproteobacteria on a separate branch adjacent to members of the genera Reinekea and Kangiella, sharing less than 88 % 16S rRNA gene sequence similarity with all recognized species of the Gammaproteobacteria. The major isoprenoid quinone was Q-8. Polar lipids consisted of phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and two unknown phospholipids. Fatty acid analysis revealed iso-C(15 : 0), C(16 : 1)omega7c and C(16 : 0) as the dominant components. The DNA G+C content was 43.8 mol%. Based on its unique phenotypic characteristics and phylogenetic remoteness, marine isolate KMM 3900(T) is considered to represent a novel genus and species, for which the name Marinicella litoralis gen. nov., sp. nov. is proposed. The type strain of Marinicella litoralis is KMM 3900(T) (=NRIC 0758(T) =JCM 16154(T)).

Two bacterial strains, KMM 3891(T) and KMM 3892, were isolated from internal tissues of the marine mollusc Umbonium costatum collected from the Sea of Japan. The novel isolates were Gram-negative, aerobic, faint pink-reddish-pigmented, rod-shaped, non-motile, stenohaline and psychrotolerant bacteria that were unable to degrade most tested complex polysaccharides. Polar lipids consisted of phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. Fatty acid analysis revealed C(17 : 1)omega6c, C(17 : 0), C(16 : 0) and C(16 : 1)omega7c as the dominant components. The major isoprenoid quinone was Q-7. The DNA G+C content of strain KMM 3891(T) was 51.7 mol%. According to phylogenetic analysis of 16S rRNA gene sequences, strains KMM 3891(T) and KMM 3892 were positioned within the Gammaproteobacteria as a separate branch, sharing <93 % sequence similarity to their phylogenetic relatives including Saccharophagus degradans, Microbulbifer species, Endozoicomonas elysicola, Simiduia agarivorans and Teredinibacter turnerae. Based on phenotypic characterization and phylogenetic distance, the novel marine isolates KMM 3891(T) and KMM 3892 represent a new genus and species, for which the name Umboniibacter marinipuniceus gen. nov., sp. nov. is proposed. The type strain of Umboniibacter marinipuniceus is KMM 3891(T) (=NRIC 0753(T) =JCM 15738(T)).

The phenotypic and phylogenetic characteristics of an aerobic, Gram-negative, motile, non-pigmented Alteromonas-like bacterium (designated strain KMM 3894(T)), isolated from a sandy sediment sample collected offshore of the Sea of Japan, were investigated. Comparative 16S rRNA gene sequence analysis revealed that strain KMM 3894(T) belonged to the genus Aestuariibacter and was most closely related to Aestuariibacter halophilus JC2043(T) (95.5 % sequence similarity). Fatty acid analysis showed C(16 : 1)omega7c, C(18 : 1)omega7c, and C(16 : 0) as the dominant components. Strain KMM 3894(T) could be differentiated from recognized species of the genus Aestuariibacter by its ability to grow at 4 degrees C and at 30 degrees C, the optimum temperature for growth, and its inability to utilize most carbohydrates. On the basis of the phenotypic, chemotaxonomic and phylogenetic data, strain KMM 3894(T) is considered to represent a novel species of the genus Aestuariibacter, for which the name Aestuariibacter litoralis sp. nov. is proposed. The type strain is KMM 3894(T) (=NRIC 0754(T)=JCM 15896(T)).

A Marinomonas-like bacterium, strain KMM 3893(T), was isolated from a marine sandy sediment collected close to shore from the Sea of Japan and subjected to a phenotypic and phylogenetic study. Comparative 16S rRNA gene sequence analysis confirmed the novel strain's assignment to the genus Marinomonas. Strain KMM 3893(T) constituted a separate phyletic line in the genus Marinomonas, sharing <97 % sequence similarity with respect to other recognized Marinomonas species. Chemotaxonomically, strain KMM 3893(T) contained the predominant fatty acids C(18 : 1)omega7c, C(16 : 1)omega7c and C(16 : 0) and had a DNA G+C content of 50.0 mol%. On the basis of the phylogenetic analysis and physiological and biochemical characterization, strain KMM 3893(T) represents a novel species of the genus Marinomonas, for which the name Marinomonas arenicola sp. nov. is proposed. The type strain is KMM 3893(T) (=NRIC 0752(T) =JCM 15737(T)).

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)).

Two novel Psychrobacter-like bacteria, strains KC 40(T) and KC 65, were isolated from a marine crustacean specimen collected from the Sea of Japan, and were characterized by using a polyphasic approach. Strains were selected on the basis of their ability to produce black-brown diffusible pigments on commonly used organic media, which appears to be a unique characteristic of recognized members of the genus Psychrobacter. Phylogenetic analyses based on both 16S rRNA and gyrB gene sequences showed that the novel isolates formed a separate cluster within the genus Psychrobacter. Strains KC 40(T) and KC 65 shared highest levels of 16S rRNA gene sequence similarity with Psychrobacter urativorans DSM 14009(T) (98.0 %), Psychrobacter pulmonis CCUG 46240(T) (97.9 %), Psychrobacter cibarius JG-219(T) (97.9 %), Psychrobacter faecalis Iso-46(T) (97.8 %), Psychrobacter aquimaris SW-210(T) (97.6 %), Psychrobacter namhaensis SW-242(T) (97.6 %) and Psychrobacter nivimaris 88/2-7(T) (97.6 %). DNA-DNA hybridization experiments revealed 84 % DNA-DNA relatedness between strains KC 40(T) and KC 65 but much lower levels of relatedness (7-35 %) between the novel strains and the type strains of recognized Psychrobacter species, confirming their assignment to a single novel species of the genus Psychrobacter. The two novel strains could be distinguished from recognized species of the genus Psychrobacter based on a combination of physiological and biochemical characteristics. On the basis of phenotypic and molecular properties, strains KC 40(T) and KC 65 are considered to represent a novel species of the genus Psychrobacter, for which the name Psychrobacter fulvigenes sp. nov. is proposed. The type strain is KC 40(T) (=KMM 3954(T)=NRIC 0746(T)=JCM 15525(T)).

A Sphingomonas-like bacterium, strain KC7(T), was isolated from a marine crustacean specimen obtained from the Sea of Japan and subjected to a polyphasic study. Comparative 16S rRNA gene sequence analysis positioned the novel strain in the genus Sphingomonas as an independent lineage adjacent to a subclade containing Sphingomonas trueperi LMG 2142(T), Sphingomonas pituitosa EDIV(T) and Sphingomonas azotifigens NBRC 15497(T). Strain KC7(T) shared highest 16S rRNA gene sequence similarity (96.1 %) with S. trueperi LMG 2142(T), Sphingomonas dokdonensis DS-4(T) and S. azotifigens NBRC 15497(T); similarities to strains of other recognized Sphingomonas species were lower (96.0-93.9 %). The strain contained sphingoglycolipid and the predominant fatty acids were C(16 : 1), C(16 : 0) and C(18 : 1); 2-OH C(14 : 0) was the major 2-hydroxy fatty acid. Previously, these lipids have been found to be characteristic of members of the genus Sphingomonas sensu stricto. On the basis of phylogenetic analysis and physiological and biochemical characterization, strain KC7(T) represents a novel species of the genus Sphingomonas, for which the name Sphingomonas japonica sp. nov. is proposed. The type strain is KC7(T) (=KMM 3038(T) =NRIC 0738(T) =JCM 15438(T)).

An aerobic, Gram-negative, non-pigmented, non-motile bacterium, KMM 3858(T), was isolated from a sea-ice sample collected from Peter the Great Bay of the Sea of Japan, Russia, and subjected to a phenotypic and phylogenetic study. Comparative analyses based on the 16S rRNA and recA gene sequences placed strain KMM 3858(T) within the genus Pseudochrobactrum. The major chemotaxonomic characteristics were found to be the presence of phosphatidylethanolamine, phosphatidylmonomethylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, an unknown aminolipid and phosphatidylcholine, major fatty acids C(18 : 1)omega7c and C(19 : 0) cyclo, and ubiquinone Q-10, confirming the affiliation of strain KMM 3858(T) to the genus Pseudochrobactrum. On the basis of the phylogenetic analysis and the physiological and biochemical characterization, strain KMM 3858(T) should be classified as representing a novel species of the genus Pseudochrobactrum, for which the name Pseudochrobactrum glaciei sp. nov. is proposed. The type strain is strain Pi26(T) (=KMM 3858(T)=NRIC 0733(T)=JCM 15115(T)).

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.

An aerobic, Gram-negative, motile, non-pigmented bacterium, strain KMM 3042(T), isolated from a deep-sea brittle star in the Fiji Sea, was subjected to a polyphasic taxonomic study. Phylogenetic analysis based on 16S rRNA gene sequences revealed marine Mn(II)-oxidizing isolate S185-2B as the closest neighbour of strain KMM 3042(T) (99.9 % sequence similarity). The two strains formed a distinct lineage within the genus Pseudomonas adjacent to the members of the Pseudomonas borbori cluster, sharing highest sequence similarity of 97.4 and 97.0 %, respectively, with P. borbori DSM 17834(T) and Pseudomonas flavescens DSM 12071(T). The DNA-DNA hybridization value (71 %) between strains KMM 3042(T) and S185-2B confirmed their assignment to the same species. On the basis of phylogenetic analysis, DNA-DNA hybridization and physiological and biochemical characterization, strains KMM 3042(T) and S185-2B should be assigned to a novel species of the genus Pseudomonas, for which the name Pseudomonas marincola sp. nov. is proposed. The type strain is KMM 3042(T) (=NRIC 0729(T) =JCM 14761(T)).

The marine bacterium "Pseudoalteromonas januaria" SUT 11 isolated from a seawater sample produced the rare cell-bound cyclic lipodepsipeptides A/A', B/B', and C/C'. The matrix-assisted laser desorption/ionization mass spectra indicated that one bromine atom presented in the peptides B/B' and C/C', whereas the component A/A' contained no bromine atom. The acyldepsipeptides A/A'-C/C' have an identical amino acid sequence, Thr-Val-Asn-Asn-Leu/allo-Ile, but differed in C-terminal amino acid and acyl moieties. Peptides A-C have Leu as a C-terminal amino acid, whereas peptides A'-C' have allo-Ile. Acyl moieties in peptides A/A', B/B', and C/C' have been found to consist of 11-(4'-hydroxyphenyl)-undeca-2,4,6,8,10-pentaenic acid, 9-(3'-bromo-4'-hydroxyphenyl)-nona-2,4,6,8-tetraenic acid, and 11-(3'-bromo-4'-hydroxyphenyl)-undeca-2,4,6,8,10-pentaenic acid, respectively. The structure of a main pair of peptides B/B' with molecular masses 843/845 Da has been determined by means of ultraviolet, infrared, and two-dimensional nuclear magnetic resonance spectroscopy. We have demonstrated that tandem nano-electrospray ionization mass spectrometry is a very efficient way for the fast and sensitive investigation of lipopeptides A/A' and C/C' with molecular masses 791 and 869/871 Da, respectively, which have been isolated in small amounts.

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)).

An aerobic, Gram-negative, non-motile, yellow-pigmented bacterium, strain KMM 3046(T), was isolated from a deep-sea brittle star from the Fiji Sea and was subjected to a polyphasic taxonomic analysis. Strain KMM 3046(T) grew at 5-32 degrees C and in the presence of 1-12 % (w/v) NaCl. It contained MK-6 as the predominant menaquinone and 3-OH i16 : 0, 3-OH i17 : 0 and 3-OH a17 : 0 as the major fatty acids. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain KMM 3046(T) forms a distinct evolutionary lineage within the family Flavobacteriaceae (phylum Bacteroidetes), displaying 92.3-91.9 % sequence similarity with respect to Salegentibacter species. On the basis of the phenotypic and phylogenetic data, strain KMM 3046(T) represents a novel genus and species of the family Flavobacteriaceae, for which the name Marixanthomonas ophiurae gen. nov., sp. nov. is proposed. The type strain of Marixanthomonas ophiurae is KMM 3046(T) (=NRIC 0684(T)=JCM 14121(T)).

An aerobic, Gram-negative, yellow-pigmented, non-motile bacterium, designated strain KMM 3882T, was isolated from a marine bivalve (Anadara broughtoni) collected from Peter the Great Bay, Sea of Japan, and was subjected to phenotypic and phylogenetic analyses. Strain KMM 3882T was found to exert a remarkable inhibitory activity against a number of Gram-positive micro-organisms. Phylogenetic analysis based on 16S rRNA gene sequences placed strain KMM 3882T within the genus Sphingomonas, as an independent lineage adjacent to Sphingomonas dokdonensis DS-4T and Sphingomonas panni DSM 15761T. Strain KMM 3882T showed the highest 16S rRNA gene sequence similarity to Sphingomonas dokdonensis DS-4T (97.3 %); similarities of 96.5-96.7 % were obtained with Sphingomonas pituitosa DSM 13101T, Sphingomonas azotifigens NBRC 15497T, Sphingomonas asaccharolytica NBRC 15499T, Sphingomonas trueperi DSM 7225T and Sphingomonas panni DSM 15761T. Chemotaxonomically, strain KMM 3882T contained sphingoglycolipid, C(16 : 0) and C(18 : 1) as predominant fatty acids and 2-OH C(14 : 0) as a major 2-hydroxy fatty acid, confirming the affiliation of strain KMM 3882T with the genus Sphingomonas. On the basis of phylogenetic analysis, DNA-DNA hybridization and physiological and biochemical characterization, strain KMM 3882T should be classified as representing a novel species of the genus Sphingomonas, for which the name Sphingomonas molluscorum sp. nov. is proposed. The type strain is KMM 3882T (=An 18T=NRIC 0685T=JCM 14122T=CIP 109223T).

The taxonomic position of three novel, marine, heterotrophic, aerobic, pigmented, gliding bacteria, isolated from the green alga Ulva fenestrata in the Sea of Japan, was determined. 16S rRNA gene sequence analysis revealed that the strains belong to the genus Arenibacter. The results of DNA-DNA hybridization experiments supported by phenotypic and chemotaxonomic data showed that the isolates represent a novel species of the genus Arenibacter, for which the name Arenibacter palladensis sp. nov. is proposed. The type strain is KMM 3961T (= LMG 21972T = CIP 108849T).

Two marine, heterotrophic, aerobic, yellow-pigmented, agarolytic bacterial strains that are motile by means of gliding were isolated from the green alga Acrosiphonia sonderi and from sea water. Comparative 16S rRNA gene sequence analysis revealed an affiliation between the strains studied and the genus Formosa, a member of the family Flavobacteriaceae. The level of sequence similarity between strain KMM 3901T and Formosa algae KMM 3553T was 99.1 %. The results of DNA-DNA hybridization experiments and phenotypic analysis indicated that the strains represent a novel species of the genus Formosa, for which the name Formosa agariphila sp. nov. is proposed, with KMM 3901T (= KCTC 12365T = LMG 23005T = DSM 15362T) as the type strain. The description of the genus Formosa is emended with newly obtained data.

A Gram-negative bacterium, designated strain LMG 22510T, was isolated from water of a pharmaceutical company steam generator. The cells had a ring-like and horseshoe-shaped morphology and possessed gliding motility. Phylogenetic analysis of the 16S rRNA gene sequence showed that the strain was a member of the Flexibacter group within the phylum 'Bacteroidetes'; its nearest neighbour was Spirosoma linguale (88.8 % sequence similarity). DNA base content, fatty acid composition and biochemical characteristics were determined. Genotypic and phenotypic data indicated that strain LMG 22510T could not be assigned to any recognized genus; therefore, a novel genus and species is proposed, Larkinella insperata gen. nov., sp. nov., with LMG 22510T (= NCIMB 14103T) as the type strain.

A comparative study of the lipid composition of 26 strains (including type strains) of marine Gammaproteobacteria belonging to the genera Shewanella, Alteromonas, Pseudoalteromonas, Marinobacterium, Microbulbifer, and Marinobacter was carried out. The bacteria exhibited genus-specific profiles of ubiquinones, phospholipids, and fatty acids, which can serve as reliable chemotaxonomic markers for tentative identification of new isolates. The studied species of the genus Shewanella were distinguished by the presence of two types of isoprenoid quinones, namely, ubiquinones Q-7 and Q-8 and menaquinones MK-7 and MMK-7; five phospholipids typical of this genus, namely, phosphatidylethanolamine (PE), phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), lyso-PE, and acyl-PG; and the fatty acids 15:0, 16:0, 16:1 (n-7), 17:1 (n-8), i-13:0, and i-15:0. The high level of branched fatty acids (38-45%) and the presence of eicosapentaenoic acid (4%) may serve as criteria for the identification of this genus. Unlike Shewanella spp., bacteria of the other genera contained a single type of isoprenoid quinone: Q-8 (Alteromonas, Pseudoalteromonas, Marinobacterium, and Microbulbifer) or Q-9 (Marinobacter). The phospholipid compositions of these bacteria were restricted to three components: two major phospholipids (PE and PG) and a minor phospholipid, bisphosphatidic acid (Alteromonas and Pseudoalteromonas) or DPG (Marinobacterium, Microbulbifer, and Marinobacter). The bacteria exhibited genus-specific profiles of fatty acids.

The taxonomic position of a marine, gliding, pink-pigmented, aerobic, heterotrophic and Gram-negative bacterium was established using a polyphasic approach. 16S rRNA gene sequence analysis indicated that the strain was a member of the phylum 'Bacteroidetes' in which it occupied a separate lineage. The predominant cellular fatty acids were C15 : 0 iso, C17 : 0 iso 3-OH, summed feature 3 and summed feature 4. The DNA G+C content was 48.7 mol%. Phylogenetic evidence and the results of phenotypic, genotypic and chemotaxonomic analyses strongly support the assignment of the newly isolated bacterium as a member of a novel genus and species, for which the name Pontibacter actiniarum gen. nov., sp. nov. is proposed. The type strain is KMM 6156T (=KCTC 12367T=LMG 23027T). It is also proposed that the illegitimate names Reichenbachia and Reichenbachia agariperforans are replaced with Reichenbachiella and Reichenbachiella agariperforans, respectively.

Five heterotrophic, aerobic, halotolerant and pigmented bacterial strains with gliding motility were isolated from Antarctic sea water; one other isolate was collected from the sea urchin Strongylocentrotus intermedius in the Gulf of Peter the Great in the Sea of Japan. 16S rRNA gene sequence analysis indicated that the strains are members of the family Flavobacteriaceae, the nearest neighbour (with 97.1 % sequence similarity) being the misclassified species [Cytophaga] marinoflava. DNA-DNA hybridization experiments and chemotaxonomic and phenotypic analyses demonstrated that the six novel isolates represent a single species distinct from [C.] marinoflava. On the basis of its separate phylogenetic lineage (the nearest neighbours show 92 % sequence similarity), [C.] marinoflava is reclassified as Leeuwenhoekiella marinoflava gen. nov., comb. nov. A second species of this new genus, Leeuwenhoekiella aequorea sp. nov., is proposed for the six novel isolates, with strain LMG 22550(T) (=CCUG 50091(T)) as the type strain.

Two Gram-negative, non-fermentative, non-denitrifying, non-pigmented, rod-shaped bacteria that were motile by means of polar flagella, designated strains KMM 330(T) and KMM 331, were isolated from a deep-sea sponge specimen and subjected to a polyphasic taxonomic study. The new isolates exhibited 16S rRNA gene sequence similarity of 99.9 %, and their mean level of DNA-DNA relatedness was 82 %. Phylogenetic analysis based on their 16S rRNA gene sequences placed the strains within the genus Pseudomonas as an independent deep clade. Strain KMM 330(T) shared highest sequence similarity (96.3 %) with each of Pseudomonas fulva NRIC 0180(T), Pseudomonas parafulva AJ 2129(T) and Pseudomonas luteola IAM 13000(T); sequence similarity to other recognized species of the genus Pseudomonas was below 95.7 %. The marine sponge isolates KMM 330(T) and KMM 331 could be distinguished from the other recognized Pseudomonas species based on a unique combination of their phenotypic characteristics, including growth in 8 or 10 % NaCl, the absence of pigments, the inability to denitrify and lack of carbohydrate utilization. On the basis of phylogenetic analysis, physiological and biochemical characterization, strains KMM 330(T) and KMM 331 should be classified as a novel species of the genus Pseudomonas, for which the name Pseudomonas pachastrellae sp. nov. is proposed. The type strain is KMM 330(T) (=JCM 12285(T)=NRIC 0583(T)=CCUG 46540(T)).

A novel marine bacterium isolated from the soft coral Paragorgia arborea in the Sea of Okhotsk was studied by using a polyphasic taxonomic approach. The strain, KMM 6029T, was strictly aerobic, heterotrophic, yellow-pigmented, non-motile by gliding, Gram-negative and oxidase-, catalase- and alkaline phosphatase-positive. From results of 16S rRNA gene sequence analysis, strain KMM 6029T occupied a distinct lineage within the family Flavobacteriaceae and showed 95.5 % similarity to its closest relative, Formosa algae. The DNA G+C content was 37.6 mol%. Major respiratory quinone was MK-6. The predominant fatty acids were i15 : 0, a15 : 0, i15 : 1, a15 : 1, i16 : 1, i16 : 0, i16 : 0 3-OH and summed feature 3 (i15 : 0 2-OH and/or 16 : 1omega7c). On the basis of phenotypic, chemotaxonomic, genotypic and phylogenetic characteristics the novel bacterium has been assigned to Bizionia gen. nov., as Bizionia paragorgiae gen. nov., sp. nov. The type strain is KMM 6029T (=KCTC 12304T=LMG 22571T).

A novel marine bacterium, strain KMM 6050T, was isolated from the sea urchin Strongylocentrotus intermedius, which inhabits the Sea of Japan. The strain studied was strictly aerobic, heterotrophic, yellow-orange-pigmented, motile by gliding, Gram-negative and oxidase-, catalase-, beta-galactosidase- and alkaline phosphatase-positive. The results of 16S rRNA gene sequence analysis showed that strain KMM 6050T occupies a distinct lineage within the family Flavobacteriaceae and is most closely related to the species Mesonia algae and Salegentibacter salegens (sequence similarity of 92.5-92.6 %). The DNA G+C content of KMM 6050T was 39.6 mol%. The major respiratory quinone was MK-6. The predominant fatty acids were i15 : 0, a15 : 0, 15 : 0, i16 : 1, i16 : 0, i16 : 0 3-OH and i17 : 0 3-OH. On the basis of phenotypic, chemotaxonomic, genotypic and phylogenetic characteristics, the novel bacterium has been assigned to the genus Gramella gen. nov., as Gramella echinicola sp. nov. The type strain is KMM 6050T (=KCTC 12278T=NBRC 100593T=LMG 22585T).

Three novel heterotrophic, Gram-negative, yellow-pigmented, aerobic, gliding, oxidase- and catalase-positive bacteria were isolated from algae collected in the Gulf of Peter the Great, Sea of Japan. 16S rRNA gene sequence analysis revealed that the strains studied represented members of the family Flavobacteriaceae and showed 93.5-93.8 % similarity with their closest relative, Psychroserpens burtonensis. The DNA G+C content of the strains was 34-37 mol%. The major respiratory quinone was MK-6. The predominant fatty acids were iso-C(15 : 0), anteiso-C(15 : 0), iso-C(15 : 1), iso-C(16 : 0)-3OH and iso-C(17 : 0)-3OH. On the basis of their phenotypic, chemotaxonomic, genotypic and phylogenetic characteristics, the newly described bacteria have been assigned to the new genus Winogradskyella gen. nov., as Winogradskyella thalassocola sp. nov. (type strain, KMM 3907T=KCTC 12221T=LMG 22492T=DSM 15363T), Winogradskyella epiphytica sp. nov. (type strain, KMM 3906T=KCTC 12220T=LMG 22491T=CCUG 47091T) and Winogradskyella eximia sp. nov. (type strain, KMM 3944T (=KCTC 12219T=LMG 22474T).

A marine, heterotrophic, Gram-negative, aerobic, yellow-pigmented, bacterium that was motile by gliding, isolated from the green alga Acrosiphonia sonderi, was studied by polyphasic taxonomic methods. 16S rRNA gene sequence analysis indicated that strain KMM 6031T formed a distinct lineage within the family Flavobacteriaceae. On the basis of phenotypic, chemotaxonomic, genotypic and phylogenetic analyses, the novel bacterium was classified as Pibocella ponti gen. nov., sp. nov. The type strain is KMM 6031T (=KCTC 12262T=NBRC 100591T=LMG 22573T).

The taxonomic position of three novel sea-water isolates was determined. The strains studied were strictly aerobic, heterotrophic, pigmented, motile by gliding, Gram-negative and oxidase-, catalase-, beta-galactosidase- and alkaline phosphatase-positive. 16S rRNA gene sequence phylogenetic analysis indicated that the strains KMM 6020T, KMM 6021 and KMM 6028 occupied a distinct lineage within the family Flavobacteriaceae. The major respiratory quinone was MK-6. The predominant fatty acids were i15 : 0, i15 : 1, i15 : 0 3-OH, i17 : 1omega9c and i17 : 0 3-OH. On the basis of phenotypic, chemotaxonomic, genotypic and phylogenetic characteristics, the novel bacteria were assigned to the genus Aquimarina gen. nov., as Aquimarina muelleri gen. nov., sp. nov. The type strain is KMM 6020T (=KCTC 12285T=LMG 22569T). From the results of the 16S rRNA gene sequence analysis and phenotypic features, the species [Cytophaga] latercula Lewin 1969 is proposed to be reclassified in the new genus Stanierella as Stanierella latercula gen. nov., comb. nov., with type strain CIP 104806T (=ATCC 23177T=NCIMB 1399T=LMG 1343T).

The taxonomic position of a novel marine bacterium isolated from the green alga Ulva fenestrata collected in the Sea of Japan was established. Cells of the strain studied, designated KMM 6017T, were strictly aerobic, heterotrophic, pink-pigmented, non-motile by gliding, Gram-negative and oxidase-, catalase-, beta-galactosidase- and alkaline phosphatase-positive. 16S rRNA gene sequence analysis indicated that the strain occupied a distinct lineage within the phylum 'Bacteroidetes' and formed a cluster with [Flexibacter] tractuosus and Reichenbachia agariperforans. The G+C content of the DNA of KMM 6017T was 40.2 mol%. The major respiratory quinone was MK-7. The predominant fatty acids were i15 : 1, i15 : 0 and i17 : 0 3-OH (34.2, 24 and 7.7 %, respectively). On the basis of phenotypic, chemotaxonomic, genotypic and phylogenetic characteristics, the novel bacterium was assigned to the genus Roseivirga gen. nov., as Roseivirga ehrenbergii gen. nov., sp. nov. The type strain is KMM 6017T (=KCTC 12282T=LMG 22567T).

Four marine heterotrophic, aerobic, pink-pigmented and non-motile bacterial strains were isolated from sea water and algae collected in the Sea of Japan. In a polyphasic taxonomic study, 16S rRNA gene sequence analysis indicated that the strains were phylogenetically highly related to Algoriphagus ratkowskyi LMG 21435T, a member of the phylum Bacteroidetes. Further phenotypic, chemotaxonomic and genomic analyses revealed that the strains represent three novel species of the genus Algoriphagus, for which the following names are proposed: Algoriphagus aquimarinus sp. nov., Algoriphagus chordae sp. nov. and Algoriphagus winogradskyi sp. nov., with type strains KMM 3958T (= LMG 21971T = CCUG 47101T), KMM 3957T (= LMG 21970T = CCUG 47095T) and KMM 3956T (= LMG 21969T = CCUG 47094T), respectively. The species Hongiella halophila Yi and Chun 2004 is transferred to the genus Algoriphagus as Algoriphagus halophilus comb. nov. because of its close phylogenetic relatedness to Algoriphagus species and analogous phenotypic and chemotaxonomic properties. The above-mentioned novel species descriptions and species reclassification justify emended descriptions of the genera Algoriphagus and Hongiella.

Three strains of the marine, gliding, pigmented, facultatively anaerobic, heterotrophic, Gram-negative bacteria were isolated from the green algae Acrosiphonia sonderi (Kütz) Kornm and Ulva fenestrata Ruprecht inhabiting the Sea of Japan. 16S rDNA sequence analysis indicated that the strains were members of the family Flavobacteriaceae, in which they occupied separate lineages. The predominant cellular fatty acids were i15 : 0, a15 : 0, i15 : 1, 15 : 0, 15 : 1omega6c, i15 : 0 3-OH and i17 : 0 3-OH. The DNA base compositions were 31-33 mol% G+C. Based on the phenotypic, genotypic, chemotaxonomic and phylogenetic analyses, the novel bacteria should be placed in a novel taxon as Algibacter lectus gen. nov., sp. nov. with type strain KMM 3902G (=KCTC 12103T=DSM 15365T).

Six novel gliding, heterotrophic, Gram-negative, yellow-pigmented, aerobic, oxidase- and catalase-positive bacteria were isolated from the green alga Ulva fenestrata, sea water and a bottom sediment sample collected in the Gulf of Peter the Great, Sea of Japan. 16S rRNA gene sequence analysis revealed that the strains studied were members of the family Flavobacteriaceae. On the basis of their phenotypic, chemotaxonomic, genotypic and phylogenetic characteristics, the novel bacteria have been assigned to the new genus Maribacter gen. nov., as Maribacter sedimenticola sp. nov., Maribacter orientalis sp. nov., Maribacter aquivivus sp. nov. and Maribacter ulvicola sp. nov., with the type strains KMM 3903T (=KCTC 12966T=CCUG 47098T), KMM 3947T (=KCTC 12967T=CCUG 48008T), KMM 3949T (=KCTC 12968T=CCUG 48009T) and KMM 3951T (=KCTC 12969T=DSM 15366T), respectively.

Four marine bacterial strains, designated KMM 3587T, KMM 3586, KMM 3821 and KMM 3822, were isolated from the sipuncula Phascolosoma japonicum, a common inhabitant of Troitza Bay in the Gulf of Peter the Great (Sea of Japan region), and from an unidentified hydrocoral species collected in Makarov Bay (Iturup Islands), Kuril Islands, North-West Pacific Ocean. The strains were characterized to clarify their taxonomic position. 16S rRNA gene sequences of KMM 3587T and KMM 3586 indicated 99% similarity to Shewanella colwelliana. Despite such a high level of 16S rRNA gene sequence similarity, DNA-DNA hybridization experiments demonstrated only 45-52% binding with DNA of S. colwelliana ATCC 39565T. The DNA G+C contents of the novel strains were 45 mol% and the shared level of DNA hybridization was conspecific (81-97%), indicating that they represent a single genospecies. The novel strains were mesophilic (able to grow at 10-34 degrees C), neutrophilic and haemolytic, and able to degrade gelatin, casein and Tween 20, 40 and 80, but not starch, agar, elastin, alginate or chitin. The major fatty acids were i13 : 0, i15 : 0, 16 : 0, 16 : 1omega7 and 17 : 1omega8 (68.9% of total). The major isoprenoid quinones were Q7 (47-62%) and Q8 (26-47%). Eicosapentaenoic acid was produced in minor amounts. Based on these data, the strains are assigned to a novel species, Shewanella affinis sp. nov. (type strain KMM 3587T=CIP 107703T=ATCC BAA-642T).

Two heterotrophic, aerobic, Gram-negative, pigmented and non-motile marine bacteria that were isolated from the green alga Ulva fenestrata were studied by polyphasic taxonomic methods. 16S rDNA sequence analysis indicated that strain KMM 3912T formed a distinct lineage within the family Flavobacteriaceae. On the basis of phenotypic, chemotaxonomic, genotypic and phylogenetic analyses, the novel bacteria were classified as Ulvibacter litoralis gen. nov., sp. nov. The type strain is KMM 3912T (=KCTC 12104T=CCUG 47093T).

The taxonomic position of four heterotrophic, aerobic, Gram-negative, non-motile and moderately halophilic marine bacteria, isolated from the green alga Acrosiphonia sonderi (Kütz) Kornm, was established. 16S rDNA sequence analysis indicated that the strains studied are members of the family Flavobacteriaceae, in which they form a distinct lineage. On the basis of phenotypic, chemotaxonomic, genotypic and phylogenetic data, the novel bacteria were classified as Mesonia algae gen. nov., sp. nov. The type strain is KMM 3909(T) (=KCTC 12089(T)=CCUG 47092(T)).

Although bacteria of the genus Shewanella belong to one of the readily cultivable groups of "Gammaproteobacteria", little is known about the occurrence and abundance of these microorganisms in the marine ecosystem. Studies revealed that of 654 isolates obtained from marine invertebrates (ophiuroid Amphiopholis kochii, sipuncula Phascolosoma japonicum, and holothurian Apostichopus japonicus, Cucumaria japonica), seawater and sediments of the North-West Pacific Ocean (i.e. the Sea of Japan and Iturup Is, Kurile Islands), 10.7% belonged to the genus Shewanella. The proportion of viable Shewanella species varied from 4% to 20% depending on the source of isolation. From the isolation study, representative strains of different phenotypes (from seventy presumptive Shewanella strains) were selected for detailed characterization using phenotypic, chemotaxonomic, and phylogenetic testing. 16S rDNA sequence-based phylogenetic analysis confirmed the results of tentative identification and placed the majority of these strains within only a few species of the genus Shewanella with 98-99% of 16S rDNA sequences identity mainly with S. japonica and S. colwelliana, suggesting that the strains studied might belong to these species. Numerically dominant strains of S. japonica were metabolically active and produced proteinases (gelatinases, caseinases), lipases, amylases, agarases, and alginases. Shewanella strains studied demonstrated weak antimicrobial and antifungal activities that might be an indication of their passive role in the colonization on living and non-living surfaces.

A promising producer of extracellular amylases, Aspergillus flavipes, was selected from 245 strains of marine fungi. Depending on the conditions of growth, this strain produced diverse amylolytic complexes. When grown on medium containing peptone and yeast extract (pH 7.0), A. flavipes synthesized three forms of amylase, differing in pH optimum (5.5, 6.0, and 7.5). A single form of the enzyme was synthesized either in the absence of peptone from the medium or at the initial pH value of the medium, equal to 8.6. The activity of the isolated amylase forms decreased in the presence of proteolytic enzymes. New, highly stable forms of amylase (with pH optima of 5.5 and 7.5 and maximum activity at 60-80 degrees C) were synthesized in the presence of diisopropyl fluorophosphate, an inhibitor of proteases.

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).

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).

The cellular phospholipids (PLs) and fatty acids (FAs) were investigated in type and environmental strains of Pseudoalteromonas, Alteromonas macleodii, A. infernus, and in three type strains of Marinomonas, M. communis, M. vaga, M. mediterranea. A total of 40 strains (19 strains in this study and 21 reported in previous papers), including Idiomarina abyssalis, I. zobellii, and Glaciecola punicea, G. pallidula, aerobic Alteromonas-like proteobacteria showed genus-characteristic patterns of phospholipids and fatty acids useful for genera discrimination. The PL patterns of surface cultures of alteromonads, pseudoalteromonads, and marinomonads consisted almost entirely of phosphatidyl ethanolamine and phosphatidyl glycerol presented in different proportions. Neither diphosphatidyl glycerol nor glycophospholipids were found in bacteria studied. In addition, the minor amount of a glycolipid was found in all strains studied. Bacteria of the genera Marinomonas, Idiomarina, and Glaciecola were clearly distinguished by presence of one of the major FAs: 18:1 (n-7), i15:0, and 16:1 (n-7), respectively. The amounts of these FAs reached up to 40-60% of total FAs. Members of Alteromonas and Pseudoalteromonas were characterized by different ratio of the following major FAs:16:1(n-7), 16:0, 17:1 (n-8), and 18:1 (n-7).

The study of the phospholipid composition of 14 type strains of marine proteobacteria of the genus Pseudoalteromonas showed that phospholipids are the main polar lipid constituents of membranes in these proteobacteria. The phospholipid patterns of the strains studied were found to be similar and involved five phospholipids typical of gram-negative bacteria, namely, phosphatidylethanolamine, phosphatidylglycerol, bisphosphatidic acid, lysophosphatidylethanolamine, and phosphatidic acid. The major phospholipids were phosphatidylethanolamine and phosphatidylglycerol, which add up to 89-97% of total phospholipids; bisphosphatidic acid was dominant among minor phospholipids. The prevalence of phosphatidylethanolamine (62-77% of total phospholipids) and the absence of diphosphatidylglycerol are the characteristic features of most bacteria of this genus. As in Escherichia coli, the phospholipid composition of the marine proteobacteria depended on the presence of magnesium in the medium.

A competitive ELISA using the intact toxin as a coating antigen for detecting palytoxin was developed. This immunoassay allows palytoxin (PTX) to be determined in the range of 6-250 ng/ml. In sensitivity, this determination is comparable with RIA but is three times inferior to ELISA using monoclonal antibodies. Inhibition experiments using some toxins of marine invertebrates proved the serological specificity of the palytoxin binding to antibodies. Both the indirect and competitive ELISA were used to find PTX-producing bacteria among 420 isolates of sea bacteria. It was found that gram-negative bacteria Aeromonas sp. and Vibrio sp. associated with toxic samples of the soft coral Palythoa sp. produced compounds antigenically related to PTX.

The 16S rRNA gene of Alteromonas distincta KMM 638T was amplified, cloned and sequenced. The nucleotide sequence was aligned with sequences of representative strains of Alteromonas, Moritella, Pseudoalteromonas and Shewanella. Results of phylogenetic analysis, using neighbour-joining and Fitch-Margoliash methods, clearly indicated that this species should be assigned to the genus Pseudoalteromonas. On the basis of polyphasic data obtained from previous work and this study, it is proposed that the species Alteromonas distincta be reclassified as Pseudoalteromonas distincta comb. nov. with type strain KMM 638T (= ATCC 700518T).

Y. pseudotuberculosis strains were grown at 6 degrees-8 degrees C and then incubated at 37 degrees C. 3-6 hours later serum resistance appeared in the strains having plasmid virulence and producing outer membrane polypeptide with a molecular weight of 120 kD, known as P1. 10-12 hours later serum resistance appeared in the strain having the virulence plasmid, but not producing P1, as well as in strains in which the plasmid was eliminated.

The protein moiety of endotoxin from Yersinia pseudotuberculosis was found to consist of two polypeptides with apparent molecular masses 40 and 14.5 kDa (4:1 w/w). The major protein (40 kDa) was isolated from the endotoxin pretreated with sodium deoxy cholate by gel chromatography on the Sephadex G-200 column. Comparative study of this protein and oligomeric form of porin from the outer membrane of Y. pseudotuberculosis using SDS--PAGE, velocity sedimentation, lipid bilayer experiments, chemical and serological analyses revealed their identity. The deoxycholate treatment of the endotoxin does not affect complexes of the major protein and LPS.