Structure of the O-specific polysaccharide from Pseudoalteromonas elyakovii sp. nov. CMM 162.

The structure of the O-specific polysaccharide from lipopolysaccharide of Pseudoalteromonas elyakovii sp. nov. CMM 162 on the basis of NMR data, Smith degradation and methylation study was elucidated as follows:-->2)-alpha-D-Glcp-(1-->4)-beta-D-GalpNAc-(1--> 3)-alpha-D-Galp-(1-->3)-beta-D-Galp-NAc-(1-->6)-alpha-D-Glcp-(1-->.

Structure of the acidic polysaccharide chain of the lipopolysaccharide of Shewanella alga 48055.

A lipopolysaccharide (LPS) with an acidic polysaccharide chain was isolated from the bacterium Shewanella alga strain 48055 and cleaved selectively at the glycosidic linkage of N-acetylneuraminic acid to give a tetrasaccharide. Studies of the tetrasaccharide and the O-deacylated LPS by 1H and 13C NMR spectroscopy, including 2D COSY, TOCSY, NOESY, rotating-frame NOE spectroscopy (ROESY), and H-detected 1H, 13C heteronuclear multiple-quantum coherence (HMQC) experiments, revealed the following structure of the polysaccharide repeating unit: -->3)-beta-D-GalpA6GroN-(1-->3)-beta-D-GlcpNAc-(1-->3)-alpha-D- GalpA6GroN- (1-->4)-alpha-Neup5Ac-(2--> where GroN is an amidically linked residue of 2-amino-1,3-propanediol (2-amino-2-deoxyglycerol). A similar structure, but with 2-acetamido-2,6-dideoxy-D-glucose instead of 2-acetamido-2-deoxy-D-glucose, has been reported previously for the polysaccharide chain of a non-O1 Vibrio cholerae H11 LPS [E. V. Vinogradov, O. Holst, J.E. Thomas-Oates, K.W. Broady, and H. Brade, Eur. J. Biochem., 210 (1992) 491-498].

[Elucidation of structure of lipid A from the marine Gram-negative bacterium Pseudoalteromonas haloplanktis ATCC 14393T]. / Ustanovlenie struktury lipida A iz morskoi gramotritsatel'noi bakterii Pseudoalteromonas haloplanktis ATSS 14393T.

The chemical structure of lipid A from the marine gamma-proteobacterium Pseudoalteromonas haloplanktis ATCC 14393T, a main product of lipopolysaccharide hydrolysis (1% AcOH), was determined using chemical methods and NMR spectroscopy. The lipid A was shown to be beta-1,6-glucosaminobiose 1,4'-diphosphate acylated with two (R)-3-hydroxyalkanoic acid residues at C3 and C3' and amidated with one (R)-3-hydroxydodecanoyl and one (R)-3-dodecanoyloxydodecanoyl residue at N2 and N2', respectively.

[Structure of glycerophosphate-containing O-specific polysaccharide from Pseudoalteromonas sp. KMM 639]. / Struktura glitserofosfatsoderzhashchego O-spetsificheskogo polisakharida Pseudoalteromonas sp. KMM 639.

On the basis of acid hydrolysis, dephosphorylation, methylation, and 13C NMR spectroscopy data, the O-specific polysaccharide of Pseudoalteromonas sp. KMM 639 was shown to be a glycerophosphate-containing polymer built of repeating disaccharide units of the following structure: [formula: see text]

[Microbiological monitoring of causative agents of sapronoses in the water of the Bogatinskoe water reservoir]. / Mikrobiologicheskii monitoring vozbuditelei sapronozov v bode Bogatinskogo vodokhranilishcha.

The results of the microbiological monitoring of potential causative agents of sapronoses in the water of the Bogatinskoye reservoir revealed that in the summer period of 1998 the mass accumulation of virulent Aeromonas sobria (up to 25% of the total number of heterotrophic bacteria) took place. The autumn period was characterized by a decrease in the number of A. sobria and the detection of bacteria of the genus Vibrio (up to 22%) with V. mimicus and V. metschnikovii identified among them in the water ecosystems of the southern regions of the Maritime Territory.

[Lipid composition of novel Shewanella species isolated from far Eastern seas].

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.

Characterization of Aeromonas and Vibrio species isolated from a drinking water reservoir.

AIMS: To study the phenotypic and chemotaxonomic (i.e. phospholipid and cellular fatty acid composition) characteristics of environmental Aeromonas spp. and Vibrio spp. isolated from a drinking water reservoir near Vladivostok City, and the application of some chemotaxonomic markers for discrimination of the two genera and species. METHODS AND RESULTS: Presumptive Aeromonas species were dominant in surface water samples (up to 25% of the total number of bacteria recovered). These strains were consistent with respect to the cultural and biochemical properties used to define the species Aeromonas sobria (seven strains) and Aer. popoffii (three strains). Vibrio mimicus (two strains) and Vibrio metschnikovii (one strain) were identified according to phenotypic features and cellular fatty acid composition. CONCLUSION: Environmental Aer. sobria isolates were atypical in their ability to grow at 42 degrees C, and were haemolytic, proteolytic and cytotoxic. Although it was present in a high proportion in the water samples, atypical Aer. sobria is not an indicator of polluted water. SIGNIFICANCE AND IMPACT OF THE STUDY: The incidence of Aeromonas in the drinking water reservoirs in the Far East of Russia is reported for the first time.

[Features of phospholipid composition of marine proteobacteria of Pseudoalteromonas species]. / Osobennosti fosfolipidnogo sostava morskikh proteobakterii roda Pseudoalteromonas.

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.

[Distribution of alpha-N-acetylgalactosaminidases in marine and freshwater microorganisms]. / Rasprostranenie alpha-N-atsetilgalaktozaminidaz sredi morskikh i presnovodnykh mikroorganizmov.

The distribution of the alpha-N-acetylgalactosidases producers among 856 marine bacteria isolated from sea water, sediments and associated with sponges, mollusks, echinoderms inhabited in different area of the Pacific Ocean and 423 Baikalian microorganisms from fresh water and sponges have been studied. About 24% as marine as Baikalian strains produced this enzyme. The most number of alpha-N-Ac-galactosidase producers were found for marine bacteria genera Aeromonas, Alteromonas and Vibrio (71% and by 60%, respectively) and Baikalian microorganisms genera Aeromonas and Bacillus (40% and 31%, respectively). alpha-N-Acetyl-galactosidase was not found for marine arthrobacters, Flexibacter-Cytophaga and baikalian Micrococcus and Rhodococcus. The promising sources of the alpha-N-Ac-galactosidases were found among Alteromonas and Bacillus.

Structure of lipid A from the marine gamma-proteobacterium Marinomonas vaga ATCC 27119T lipopolysaccharide.

The chemical structure of a novel lipid A obtained as a major component on hydrolysis of LPS from the marine gamma-proteobacterium Marinomonas vaga ATCC 27119T with 1% AcOH was determined. Using chemical analysis and NMR data, it was shown to be beta-1,6-glucosaminobiose 1-phosphate acylated with R-3-hydroxydecanoic acid (at position 3), and R-3-dodecanoyloxydecanoic (or R-3-decanoyloxydecanoic) acid and R-3-(R-3-hydroxydecanoyl)oxydecanoic acids (at the 2- or 2;-positions). The absence of a fatty acid at the 3;-position and a phosphoryl group at the 4;-position, and also the presence of R-3-acyloxyalkanoic acid with R-3-hydroxyalkanoic acid as the secondary acid are unique features distinguishing the M. vaga lipid A from other ones.

Structure of O-specific polysaccharide from Pseudoalteromonas nigrifaciens strain KMM 161.

On mild acid degradation of the lipopolysaccharide of the marine microorganism Pseudoalteromonas nigrifaciens KMM 161 an O-specific polysaccharide containing D-galactose, 2-acetamido-2-deoxy-D-glucose, 3,6-dideoxy-3-(4-hydroxybutyramido)-D-galactose, and 2-acetamido-2-deoxy-L-guluronic acid residues was obtained. From the results of Smith degradation, O-deacetylation of the polysaccharide, and NMR spectroscopy the following structure of the tetrasaccharide repeating unit of the O-specific polysaccharide was established [see reaction]. It should be noted that the same structure occurs in the antigenic polysaccharide of Pseudoalteromonas nigrifaciens KMM 158 described earlier as Alteromonas macleodii 2MM6.

[Diversity in the monosaccharide composition of antigenic polysaccharides from proteobacteria Pseudoalteromonas and Marinomonas genera]. / Raznoobrazie monosakharidnogo sostava antigennykh polisakharidov proteobakterii rodov Pseudoalteromonas i Marinomonas.

The sugar analysis of the glycans of the type strains of marine proteobacteria of the genera Pseudoalteromonas and Marinomonas--Pseudoalteromonas atlantica IAM12927T, P. aurantia NCIMB 2033T, P. citrea ATCC 29719T, P. elyakovii KMM 162T, P. espejiana ATCC 29659T, P. piscicida NCIMB 645T, P. tetraodonis IAM 14160T, Marinomonas communis ATCC 27118T, and M. vaga ATCC 27119T--showed that they contain glucose, galactose, galactosamine, glucosamine, fucose, rhamnose, mannose, heptose, 2-keto-3-deoxyoctonate (KDO), uronic acids, colitose (3,6-dideoxyl-L-xylo-hexose), and 6-deoxy-L-talose. The carbohydrate composition of the antigenic polysaccharides (PSs) of P. elyakovii KMM 162T and P. espejiana ATCC 29659T depended on the type and the concentration of carbohydrate substrates in the nutrient media. The molar proportion between rhamnose, glucose, and galactose (ca. 1:0.3:2) in the PS of P. elyakovii KMM 162T was almost the same in the media lacking carbohydrates or containing glucose or galactose at a concentration of 1 g/l. At the same time, the molar proportion between fucose, glucose, galactose, galactosamine, and glucosamine (ca. 1:1:1:2:0.5) in the PS of P. espejiana ATCC 29659T depended on the presence and the concentration of carbohydrate substrates in the medium. A high concentration of glucose in the medium (30 g/l) brought about a rise in the content of glucose in PSs (9-fold for the PS of P. elyakovii KMM 162T and 4.6-fold for the PS of P. espejiana ATCC 29659T) and led to a decrease in the content of other carbohydrates. The cultivation of these two strains at a lactose concentration of 30 g/l resulted in their PSs containing glucose and galactose in about equal proportions (ca. 1:1 in the case of P. espejiana ATCC 29659T and ca. 2.1:1.7 in the case of P. elyakovii KMM 162T).

Evaluation of phospholipid and fatty acid compositions as chemotaxonomic markers of Alteromonas-like proteobacteria.

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).

Shewanella japonica sp. nov.

Two strains of agar-digesting bacteria, KMM 3299T and KMM 3300, respectively isolated from sea water and the mussel Protothaca jedoensis, have been characterized. Based on sequencing of the 16S rRNA gene, KMM 3299T showed the highest similarity (93-95%) to members of the genus Shewanella. The G+C contents of the DNAs of these strains were 43-44 mol%. The level of DNA homology between the two strains was conspecific (95%), indicating that they represent a distinct genospecies. These organisms were non-pigmented, Gram-negative, polarly flagellated, facultatively anaerobic, mesophilic, neutrophilic and able to degrade a wide range of high molecular mass polymers, including alginate, carrageenan, laminaran and agar. The novel organisms were susceptible to gentamycin, carbenicillin, lincomycin and oleandomycin. The predominant cellular fatty acids were i-15:0, 16:0, 16:1(n-7), 18:1(n-7). Eicosapentaenoic acid, 20:5(n-3), was detected in the two isolates at levels of 1-8%, depending on the temperature of cultivation. Phylogenetic evidence, together with phenotypic characteristics, showed that the two isolates studied constitute a novel species of the genus Shewanella. The name Shewanella japonica is proposed; the type strain is KMM 3299T(= LMG 19691T = CIP 106860T).

Characterization of Bacillus strains of marine origin.

A total of twenty aerobic endospore-forming bacilli, isolated from marine invertebrates and sea water of different areas of the Pacific Ocean, were taxonomically characterized. Most of the bacilli (11 strains) of marine origin belonged to the species Bacillus subtilis, according to their phenotypic characteristics, antibiotic susceptibility profiles, and fatty acids patterns. A group of four alkaliphilic strains formed a separate cluster that was tentatively classified as B. horti. One isolate, KMM 1717, associated with a sponge from the Coral Sea was identified as B. pumilus. Two strains, Bacillus KMM 1916 and KMM 1918, showed antibiotic sensitivity profiles similar to B. licheniformis, but they had a distinct fatty acid composition and peculiar phenotypic traits. The taxonomic affiliation of KMM 1810 and KMM 1763 remained unclear since their fatty acid composition and antibiotic sensitivity patterns were not resembled with none of these obtained for Bacillus strains.

Phenotypic diversity of Pseudoalteromonas citrea from different marine habitats and emendation of the description.

Four strains of marine, aerobic, agar-decomposing bacteria with one polar flagellum and with DNA G + C contents of 38.9-40.2 mol% were isolated from the Far-Eastern mussels Crenomytilus grayanus and Patinopecten yessoensis. These four strains were identified as Pseudoalteromonas; however, they were phenotypically different from species described previously according to carbon compound utilization tests and the BIOLOG identification system. High agar-decomposing activity was found in two strains, in one of which agarase, alpha-galactosidase, pustulanase and laminarinase had been detected. The level of DNA homology of three of the strains was 70-100%. The fourth isolate was genetically less related to the others (67% DNA relatedness) and phenotypically was more distant from other members of this group; however, all four strains were assigned to a single species genotypically. DNA from the strains isolated from mussels showed 40-45% genetic relatedness with the DNA of Alteromonas atlantica, 8-36% with DNA of Pseudoalteromonas haloplanktis subsp. haloplanktis, Pseudoalteromonas haloplanktis subsp. tetraodonis, Pseudoalteromonas undina, Pseudoalteromonas nigrifaciens and Pseudoalteromonasas carrageenovora, 53% with Pseudoalteromonas elyakovii, 32-48% with marine P. nigrifaciens from mussels and 14-16% with Alteromonas macleodii. The DNA-DNA hybridization data revealed that the levels of relatedness between the strains isolated and the type strains of Pseudoalteromonas citrea and Pseudoalteromonas fuliginea described recently were significant (95-85%). These results were confirmed by serological data employing polyclonal antibodies to cell surface antigens. The strains isolated from mussels were identified as P. citrea. The hybridization data showed that the name P. fuliginea Romanenko et al. 1994 should be recognized as a junior subjective synonym of P. citrea Gauthier 1977. A notable phenotypic diversity of P. citrea which might be a reflection of their ecological habitats is discussed.

[Degradation of fucoidan by the marine proteobacterium Pseudoalteromonas citrea]. / Osobennosti degradatsii fukoidana morskimi proteobakteriiami Pseudoalteromonas citrea.

It was found that Pseudoalteromonas citrea strains KMM 3296 and KMM 3298 isolated from the brown algae Fucus evanescens and Chorda filum, respectively, and strain 3297 isolated from the sea cucumber Apostichopus japonicus are able to degrade fucoidans. The fucoidanases of these strains efficiently degraded the fucoidan of brown algae at pH 6.5-7.0 and remained active at 40-50 degrees C. The endo-type hydrolysis of fucoidan resulted in the formation of sulfated alpha-L-fucooligosaccharides. The other nine strains of P. citrea studied (including the type strain of this species), which were isolated from other habitats, were not able to degrade fucoidan.

Chemical characterization of lipid A from some marine proteobacteria.

Lipids A from type and wild strains of marine Proteobacteria belonging to Alteromonadaceae (Alteromonas (1 species), Idiomarina (1 species), and Pseudoalteromonas (8 species) genera) and Vibrionaceae (Shewanella (1 species) and Vibrio (1 species) genera) families and Marinomonas genus (1 species) were isolated by hydrolysis of their respective lipopolysaccharides with 1% acetic acid. Based on thin-layer chromatography data, the lipids A studied had low heterogeneity and generated family-specific patterns varying in numbers of bands and their chromatographic mobility. Total chemical analysis of the compounds showed that they contained glucosamine, phosphate, and fatty acids with decanoate (I. zobellii KMM 231(T) lipid A) or dodecanoate (lipids A of the other bacteria) and 3-hydroxy alkanoates as the major fatty acid components. Unlike terrestrial bacterial lipids A, lipids A of marine Proteobacteria had basically monophosphoryl (except V. fluvialis AQ 0002B lipid A with its two phosphate groups) and pentaacyl (except S. alga 48055 and V. fluvialis AQ 0002B lipids A which were found to have six residues of fatty acids per molecule of glucosamine disaccharide) structural types, low toxicity, and may be useful as potential endotoxin antagonists.