Rheinheimera japonica sp. nov., a novel bacterium with antimicrobial activity from seashore sediments of the Sea of Japan.

Two gram-negative, aerobic, brown-pigmented, motile rod-shaped bacteria KMM 9512 and KMM 9513(T) were isolated from a sediment sample collected from the Sea of Japan seashore, Russia. On the basis of the 16S rRNA gene sequence analysis, novel strains KMM 9512 and KMM 9513(T) positioned within the genus Rheinheimera (class Gammaproteobacteria) as a separate subline adjacent to Rheinheimera baltica DSM 14885(T) sharing highest gene sequence similarities of 98.6-97.6 % to their closest phylogenetic relatives, Rheinheimera muenzenbergensis LMG 27269(T), R. baltica DSM 14885(T), Rheinheimera aquimaris JCM 14331(T), Rheinheimera nanhaiensis KACC 14030(T), and Rheinheimera pacifica KMM 1406(T). Strains KMM 9512 and KMM 9513(T) belong to the same separate genospecies on the basis of the 16S rRNA gene sequence similarity (99.8 %) and their DNA relatedness to each other (89 %) and to closely related Rheinheimera species (25-53 %). The major isoprenoid quinone was Q-8, polar lipids consisted of phosphatidylserine, phosphatidylglycerol, phosphatidylethanolamine, unknown aminolipids, unknown aminophospholipids, unknown phospholipids, and unknown lipids, and major fatty acid were C16:0, C16:1 ω7c, C17:1 ω8c, C12:0 3-OH followed by C17:0 and C18:1 ω7c in both strains. Strains KMM 9512 and KMM 9513(T) revealed a remarkable antagonistic activity toward a number of gram-positive and gram-negative microorganisms. On the basis of phylogenetic analysis, DNA-DNA hybridization results, and phenotypic differences, strains KMM 9512 and KMM 9513(T) are proposed to be classified as a novel species of the genus Rheinheimera, Rheinheimera japonica sp. nov. The type strain of this species is KMM 9513(T) = NRIC 0918(T).

Composition and distribution of fatty acids in triglycerides from goat infant formulas with milk fat.

Most infant formulas use vegetable oils in place of milk fat to provide an overall fatty acid profile similar to that of breast milk. Vegetable oils have 5 to 20% saturated fatty acids in the sn-2 position of triglycerides unless they are modified by interesterification. Interesterification is increasingly used for the fat for infant formulas to raise the level of saturated fatty acids in the sn-2 position to 40 to 60%. The objective of this study was to verify an alternative approach to providing the appropriate fatty acid profile, including in the sn-2 position, for a goat infant formula. In this method, 55% of total fat was made from goat milk fat and 45% from a mixture of unmodified high oleic sunflower, canola, and sunflower oils in a ratio of 44:30:26. The fatty acid profile was measured by gas-liquid chromatography and the relative percentage of fatty acids in the sn-2 position of triglycerides was measured via partial deacylation with Grignard reagent using trimethylsilyl derivatives of monoacylglycerols. Mixing goat milk fat with vegetable oils produced a formula with a profile of essential fatty acids and a ratio of linoleic:alpha-linolenic fatty acids within the required interval of 5 to 15:1 recommended for infant formula. The proportion of palmitic acid in the sn-2 position was 31%.

[New ceramides from sea sponge Oceanapia sp].

Total ceramides containing nonbranched and iso-branched C18- and C19-phytosphingosines acylated with non-hydroxylated fatty acids were isolated from a marine sponge Oceanapia sp. The structures of these compounds were determined by HPLC, NMR, MALDI-TOF MS, and GLC-MS of the Me3Si derivatives and by chemical transformations. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2006, vol. 32, no. 3; see also http://www.maik.ru.

Identification, isolation and characterization of tetracosapolyenoic acids in lipids of marine coelenterates.

Several tetracosapolyenoic acids (TPA) were detected in lipids of different marine coelenterates. Two of these acids were isolated and their structures were confirmed by chemical and spectral methods as all-cis-6,9,12,15,18-tetracosapentaenoic and all-cis-6,9,12,15,18,21-tetracosahexaenoic acid. Their distribution among lipids of a number of species of different classes of coelenterates from the northern and tropical seas, among neutral and polar lipids of these organisms was investigated. Significant quantities of TPA were found in all of the Octacorallia species studied. In some cases the sum of TPA reaches the level of 20% of total lipid fatty acids. The fatty acid composition of different coelenterates is also discussed.

New polar steroids from the sponges Trachyopsis halichondroides and Cymbastela coralliophila.

Fractions of trisulfated trihydroxysteroids from the sponges Trachyopsis halichondroides (two different collections) and Cymbastela coralliophila have been isolated and investigated. Ten triacetates were obtained from these fractions by desulfatation/acetylation followed by chromatographic separation, and their structures have been established. Four of these derivatives--namely, triacetates of 5 alpha-pregnane-2 beta,3 alpha,6 alpha-triol (3c), 23,24-dinor-5 alpha-cholane-2 beta,3 alpha,6 alpha-triol (4c), 5 alpha-cholest-22-ene-2 beta,3 alpha,6 alpha-triol (7c), and 24-isopropyl-5 alpha-cholestane-2 beta,3 alpha,6 alpha-triol (11c)-- are described as new derivatives of natural steroids.

[The phospholipid spectrum of the blood serum in patients with chronic non-specific pulmonary diseases during rehabilitation treatment]. / Fosfolipidnyi spektr syvorotki krovi bol'nykh khronicheskimi nespetsificheskimi zabolevaniiami legkikh v protsesse vosstanovitel'nogo lecheniia.

Lipid metabolism was studied in 49 patients with chronic non-specific pulmonary diseases. The judgement about lipid metabolism was formed on the basis of changes in the content of phospholipid (PL) fractions in the patients' blood serum under the influence of health resort treatment. PL were separated by micro-thin-layer chromatography. They included five fractions: phosphatidylcholine (PC), sphingomyelin (SM), lysophosphatidylcholine (LPC), phosphatidylethanolamine (PE) and phosphatidylinositol (PI). The patients with chronic non-specific pulmonary diseases showed a significant decrease in the PC level and a rise of SM, LPC and PI. In the course of health resort treatment, lipid metabolism in the patients did not return to normal. The tendency towards normalization was observed only by the sixth month after rehabilitation treatment. The changes discovered are of diagnostic importance and can be used in the assessment of the efficacy of health resort treatment aimed at the removal of the inflammatory process in the lungs.

[Characterization of marine bacteria Pseudoalteromonas citrea degradating fucoidan]. / Kharakteristika morskikh bakterii Pseudoalteromonas citrea degradiruiushchikh fukoidan.

Two strains KMM 3296 and KMM 3298, isolated from the surface of brown algae Chorda filum and Fucus evanescens thalli and strain KMM 3297, isolated from celomic fluid of holoturian Apostichopus japonicus, were identified as Pseudoalteromonas citrea on the basis of pheno-, geno- and phylotypic features. The studied bacteria were different from other bacteria of the species as to their ability to destruct sulphated polysaccharides of brown algae, fucoidans.

[Marine fungi in sediments of the Sea of Japan (Russian territory) and their biologically active metabolites]. / Griby gruntov Iaponskogo moria (Rossiiskoe poberezh'e) i ikh biologicheskie aktivnye metabolity.

The most abundant marine fungi encountered in various regions of the Sea of Japan belong to the genera Penicillium, Aspergillus, Wardomyces, Trichoderma, Chrysosporium, and Chaetomium. Facultative marine fungi of the genera Scytalidium, Verticillium, and Oidiodendron and obligate marine fungi of the genus Dendryphiella are much less abundant. The composition of marine sediments and the anthropogenic load on them were found to influence the abundance and species diversity of fungi, as well as the occurrence of fungal strains producing hemolytically active substances. The biodiversity of mycobiota and the abundance of hemotoxin-producing fungi in marine sediments may be used to evaluate the anthropogenic load on marine biocenoses. Hemolytic compounds were produced by 57% of the fungi isolated from marine sediments. The hemolytic activity of Chaetomium spiculipilium was revealed in the fraction of the culture liquid containing extracellular fatty acids and pigments. The fatty acid composition of this marine fungus was determined.

The heptadecanoic fatty aldehyde--one of the main aldehydes of the far-eastern Bryozoa.

1. The relative content of 16:0, 17:0 and 18:0 fatty aldehydes in the lipids of eight species of the far-eastern Bryozoa was studied. 2. Heptadecanoic aldehyde is one of the main aldehydes in the seven species investigated comprising about 30% of the sum of these main bryozoan aldehydes. 3. We suggest the unusually high relative heptadecanoic aldehyde content in the lipids of Bryozoa may be helpful in settling some problems concerning their system.

[Phospholipids in the blood serum of patients with ischemic heart disease at the stage of early rehabilitation at a sanatorium]. / Fosfolipidy syvorotki krovi u bol'nykh ishemicheskoi bolezn'iu serdtsa na étape rannei sanatornoi reabilitatsii.

Lipid metabolism has been examined in coronary patients from the content of the blood serum phospholipid fractions during exposure to the health resort factors at the South of the Primorye territory. Qualitative composition of phospholipids included 5 fractions, i.e. phosphatidylcholine, sphingomyelin, lysophosphatidylcholine, phosphatidylethanolamine, and phosphatidylinosite. Decreased levels of phosphatidylcholine and elevated concentrations of lysophosphatidylcholine and sphingomyelin have been detected in the patients. No differences in the concentrations of phospholipid fractions in patients with extensive and small-focus myocardial infarctions have been observed. Patients with repeated myocardial infarctions in the history have developed drastic changes in lipid metabolism, i.e. abnormal levels of phosphatidylcholine, lysophosphatidylcholine, and sphingomyelin. Lipid metabolism parameters have not normalized in coronary patients in the course of health resort treatment.

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).

Idiomarina gen. nov., comprising novel indigenous deep-sea bacteria from the Pacific Ocean, including descriptions of two species, Idiomarina abyssalis sp. nov. and Idiomarina zobellii sp. nov.

Two bacterial strains, KMM 227T and 231T, were isolated from seawater samples collected from the north-western Pacific Ocean at a depth of 4000-5000 m and were characterized using polyphasic taxonomy. Both were Gram-negative, psychrotolerant, heterotrophic, aerobic and required NaCl for growth (0.6-15.0%). The temperature for growth was 4-30 degrees C. Both strains were rod-shaped, with a single flagellum. However, strain KMM 231T revealed a single long fimbrium. Cellular fatty acids detected in the isolates were predominantly odd-numbered and iso-branched, with 15 and 17 carbons (ca. 70%). Also present were saturated and monounsaturated straight-chain fatty acids. Results of phylogenetic analyses, employing three tree-making methods, strongly indicated that the two strains formed a distinct lineage within a clade containing the genera Alteromonas, Colwellia and Pseudoalteromonas, in the gamma-Proteobacteria. The two strains shared 16S rDNA sequence similarity of 96.9% and genomic DNA relatedness of 27%; the latter was determined by dot-blot hybridization. The strains were differentiated by the presence of fimbria, production of chitinase, ability to grow on 15% NaCl and BIOLOG profiles. Given the polyphasic evidence accumulated in this study, it is proposed that the two deep-sea isolates be classified in the genus Idiomarina gen. nov., as Idiomarina abyssalis sp. nov. (type strain is KMM 227T) and Idiomarina zobellii sp. nov. (type strain is KMM 231T).

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.

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).

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.