Phylogenetic and Functional Prokaryotic Diversity in the Hoito-Gol Mesothermal Mineral Spring (Eastern Savan Mountains. Buryat Renuhlie).

High-throughput sequencing was used for comparative analysis of microbial communities of the water and mat from the Hoito-Gol mesothermal mineral sulfide spring (Eastern Sayan Mountains, Buryat Republic). Activity of microbial communities was determined. While both spring biotopes were dominated by members of three bacterial phyla, Proteobacteria, Bacteroidetes, and Firmicutes, they differed drastically in the composition of predominant phylotypes (at the genus level). In the water, the organisms wide spread in aquatic'environments were predominant, mostly aerobic chemoorganotrophs of the generaAcinetobacter, Pe- dobacter, and Flavobacterium. In the microbial mat,;the organisms actively involved in the sulfur cycle predominated, including sulfur-reducing bacteria Sulfurospirillum, sulfate-reducing deltaproteobacteria, sulfur- oxidizing chemoautotrophic bacteria, anoxygenic phototrophic bacteria of,the phyla Chloroflexi and Chloro- bi, as well as purple bacteria belonging to the Q-, P--, and y-Proteobacteria. Microbial mats of the spring exhibited higher phylogenetic diversity compared to high-temperature mats containing photosynthetic microorganisms.

Anoxybacillus mongoliensis sp. nov., a novel thermophilic proteinase producing bacterium isolated from alkaline hot spring, central Mongolia.

A Gram reaction positive, spore-forming, facultative anaerobic bacterium belonging to the Phylum Firmicutes, was isolated from alkaline hot (80 degrees C, pH 9.8 spring Tsenher, central Mongolia. The cells were rod shaped, feebly motile, peritrichously flagellated. Strain T4 was moderately thermophilic with optimum growth at 60 degrees C. Maximum temperature for growth was between 70 and 75 degrees C; minimum temperature for growth was between 35 and 30 degrees C. Alkalitolerant, optimum pH for growth was 8.0; minimum pH for growth was between 5.0 and 5.5 and maximum was between 10.5 and 10.8. The growth was observed at NaCl concentrations of 0-5% (w/v) with the optimum at 0.2-0.5%. No growth was observed at 6% NaCl (w/v). Aerobically, the strain utilized proteinaceous substrates, organic acids and a range of carbohydrates including glucose, ribose, sucrose and xylose as well. Anaerobically, only glucose and sucrose were utilized. Strain T4T produced thermostable alkaline subtilisin-like serine proteinase. The G + C content was 44.2 mol. % (td). On the basis of 16S rRNA gene sequence similarity strain T4(T) was shown to be closely related to the members of the genus Anoxybacillus (family Bacillaceae, class "Bacilli"). DNA-DNA hybridization data revealed that strain T4T had only 38% relatedness to A. flavithermus and 28% relatedness to A. pushchinoensis. Based on its morphology, physiology, phylogenetic relationship and its low DNA-DNA relatedness values with validly published species of Anoxybacillus, it is proposed that strain T4T represents a novel species Anoxybacillus mongoliensis sp. nov., with the type strain T4(T) (=DSM 19169 = VKM 2407).

[Extracellular proteolytic activity of bacteria from soda-salt lakes of Transbaikalia].

Influence of nitrogen source on proteinases synthesis in aerobic alkalotolerant and halotolerant bacteria from soda-salt lakes of Transbaikalia was studied. Maximal accumulation of proteinases was revealed on medium with peptones. Introduction of various sources of nitrogen in the medium did not result in increase of enzyme activity in cultural liquid. It was indicated that secreting proteinases of the studied bacteria strains possess narrow substrate specificity, hydrolyze proteins and n-nitroanilide substrates have maximal activity during GlpAALpNA hydrolysis. Data of inhibitory analysis and substrate specificity of studied extracellular enzymes indicate that they belong to a class of serine proteinases of subtilisin-like type.

[Colorless sulfur bacteria Thioploca from different sites in Lake Baikal].

The colorless sulfur bacteria Thioploca spp. found in Lake Baikal are probably a marker for the influx of subterranean mineralized fluids. Bacteria act as a biological filter; by consuming sulfide in their metabolism, they detoxicate it and maintain the purity of Lake Baikal's water. The bacteria were investigated by various techniques. According to analysis of the 16S rRNA gene fragment, Thioploca sp. from Frolikha Bay, Baikal belongs to the clade of freshwater species found in Lake Biwa and Lake Constance; it is most closely related to Thioploca ingrica.

[The structure of phototrophic communities of soda lakes of the southeastern Transbaikal region].

The structure of benthic phototrophic communities of 24 soda lakes of the southeastern Transbaikal Region was studied. The physicochemical properties of the lakes were determined. The results of the cell count of anoxygenic phototrophic bacteria (APB) belonging to various groups are presented. The influence of salinity on the structure of APB communities was investigated. The APB reaction to environmental conditions was determined. Massive development of phototrophic microorganisms in the form of mats and films was observed in the majority of the investigated lakes. The APB communities were characterized by a wide diversity of species and evenness of species composition. Purple sulfur bacteria of the families Ectothiorhodospiraceae and Chromatiaceae were predominant. Purple nonsulfur bacteria of the family Rhodobacteraceae, green filamentous bacteria Oscillochloris sp., and heliobacteria were also detected. According to preliminary data, no less than 15 species of APB occur in the studied lakes. Among them, three novel genera and four species have already been described. Identification of other isolates is still in progress. The lakes make an almost continuous series of fresh, brackish, and saline water bodies, varying in their degree of mineralization. It was demonstrated that the structure of APB communities was unaffected by salinity ranging from 5 to 40 g/l. At salt concentrations of lower than 5 g/l, the level of water mineralization became a limiting factor. Experiments with the isolated cultures showed that the APB were obligately dependent on the presence of carbonate ions in the medium. They were haloalkalitolerant or haloalkaliphilic. Thus, they are well adapted to the conditions of soda lakes with a high mineralization. It was demonstrated that soda lakes of the southeastern Transbaikal Region represent a special type of habitat which harbors a peculiar autochthonous microflora and differs from both highly mineralized soda lakes and shallow saline water bodies of the sea origin.

[Biogeochemical processes in the algal-bacterial mats of the Urinskii alkaline hot spring].

The structure and production characteristics of microbial communities from the Urinskii alkaline hot spring (Buryat Republic, Russia) have been investigated. A distinctive characteristic of this hot spring is the lack of sulfide in the issuing water. The water temperature near the spring vents ranged from 69 to 38.5 degrees C and pH values ranged from 8.8 to 9.2. The total mineralization of water was less than 0.1 g/liter. Temperature has a profound effect on the species composition and biogeochemical processes occurring in the algal-bacterial mats of the Urinskii hot spring. The maximum diversity of the phototrophic community was observed at the temperatures 40 and 46 degrees C. A total of 12 species of cyanobacteria, 4 species of diatoms, and one species of thermophilic anoxygenic phototrophic bacteria, Chloroflexus aurantiacus, have been isolated from mat samples. At temperatures above 40 degrees C, the filamentous cyanobacterium Phormidium laminosum was predominant; its cell number and biomass concentration were 95.1 and 63.9%, respectively. At lower temperatures, the biomass concentrations of the cyanobacterium Oscillatoria limosa and diatoms increased (50.2 and 36.4%, respectively). The cyanobacterium Mastigocladus laminosus, which is normally found in neutral or slightly acidic hydrothermal systems, was detected in microbial communities. As the diatom concentration increases, so does the dry matter concentration in mats, while the content of organic matter decreases. The concentrations of proteins and carbohydrates reached their maximum levels at 45-50 degrees C. The maximum average rate of oxygenic photosynthesis (2.1 g C/m2 day), chlorophyll a content (343.4 mg/m2), and cell number of phototrophic microorganisms were observed at temperatures from 45 to 50 degrees C. The peak mass of bacterial mats (56.75 g/m2) occurred at a temperature of 65-60 degrees C. The maximum biomass concentration of phototrophs (414.63 x 10(-6) g/ml) and the peak rate of anoxygenic photosynthesis [0.42 g C/(m2 day)] were observed at a temperature of 35-40 degrees C.

[Extracellular proteolytic eyzymes of microscopic fungi from thermal springs of the Barguzin Valley (Northern Baikal region)].

Production of extracellular proteolytic enzymes was studied in thermophilic fungi Paecelomyces variotii and Aspergillus carneus, isolated from thermal springs of the Barguzin Valley. Protease synthesis in these fungi requires protein in the ambient medium. The composition of the enzymes secreted by A. carneus depends on the kind of carbohydrate present in the medium. The proteinase of this fungus digests synthetic substrates and gelatin (optimum pH 7.7). It belongs to neutral serine proteases. Extracellular P. variotii proteases digests gelatin (optimum pH 9.7-10.4). According to inhibitor analysis data, they are assigned to alkaline metalloproteinases and serine proteinases.

[Aerobic methanotrophic communities in the bottom sediments of Lake Baikal].

The results of the first methodical investigation into the aerobic methanotrophic communities inhabiting the bottom sediments of Lake Baikal are reported. Use of the radioisotopic method revealed methane consumption in 12 10- to 50-cm-long sediment cores. The maximum methane consumption rates (495-737 microl/(dm3 day) were recorded in sediments in the regions of hydrothermal vents and oil and gas occurrence. Methane consumption was most active in the surface layers of the sediments (0-4 cm); it decreased with the sediment depth and became negligible or absent at depths below 20 cm. The number of methanotrophic bacteria usually ranged from 100 to 1000 cells/cm3 of sediment and reached 1 million cells/cm3 in the regions of oil and gas occurrence. The 17 enrichment cultures obtained were represented mainly by morphotype II methanotrophs. Phylogenetic analysis of the enrichment cultures in terms of the amino acid sequence of the alpha subunit of the membrane-bound methane monooxygenase revealed the predominance of methanotrophs of the genus Methylocystis. The results obtained suggest the presence of an active aerobic methanotrophic community in Lake Baikal.

[The phototrophic community found in Lake Khilganta (an alkaline saline lake located in the southeastern Transbaikal region)].

The structure of the phototrophic community found in Lake Khilganta (the Agin-Buryat Autonomous Area), a shallow saline soda lake (depth, 35-45 cm; water mineralization, 45 g/l; alkalinity, 30 mg-equiv/l; pH 9.5) has been studied. The bottom of the lake is covered with a 10- to 15-mm microbial mat, whose basis is formed by the filamentous cyanobacterium Microcoleus chthonoplastes. The mat exhibits pronounced layering and contains a significant amount of minerals. Six zones, which have characteristic colors and consistencies and are composed of intermittent layers, have been identified along the vertical profile. Live phototrophic bacteria have been found in the three upper zones. The bulk of the cyanobacteria is concentrated in the upper zone. In the lower zones, the development of purple bacteria has been observed. The diurnal dynamics of the vertical distribution of phototrophic microorganisms, which results from variations in the physicochemical environmental parameters, is described. Ectothiorhodospira sp. are dominant among the anoxyphotobacteria present. Their number, determined according to the inoculation method, is 10(6)-10(7) cells/ml. The purple bacteria of the genera Allochromatium, Thiocapsa, and Rhodovulum are also present. Experiments with isolated pure cultures have shown that the anoxygenic photosynthetic bacteria of Lake Khilganta are halotolerant and alkalitolerant or alkaliphilic. In liquid enrichment cultures, at pH 9.5, the ratio of anoxyphotobacteria species is close to that observed in the lake. When the pH is increased to 10.4, it is Ectothiorhodospira, which is the most adapted to life under increased mineralization and alkalinity, that predominantly develops. Photosynthetic activity has been observed in the three upper mat zones and constitutes, on average, 1.5 g C/(m2 h); the share of anoxygenic photosynthesis accounts for 75-95% of the total productivity. The main role in sulfide oxidation belongs to the phototrophic anoxyphotobacteria and cyanobacteria. In terms of the physicochemical conditions and structure of the phototrophic community, Lake Khilganta is similar to shallow saline water bodies of marine origin. The main differences consist in the increased alkalinity and in the consequent prevalence of alkaliphilic and alkalitolerant microorganisms and in the absence of representatives of the neutrophilic group of green sulfur bacteria.

[Bacterial processes of the methane cycle in the bottom sediments of Baikal lake]. / Bakterial'nye protsessy tsikla metana v donnykh osadkakh ozera Baikal.

The activity of methanogenic and methanotrophic bacteria was evaluated in bottom sediments of Lake Baikal. Methane concentration in Baikal bottom sediments varied from 0.0053 to 81.7 ml/dm3. Bacterial methane was produced at rates of 0.0004-534.7 microliters CH4/(dm3 day) and oxidized at rates of 0.005-1180 microliters CH4/(dm3 day). Peak methane production and oxidation were observed in Frolikha Bay near a methane vent. Methane was emitted into water at rates of 49.2-4340 microliters CH4/(m2 day). Rates of bacterial methane oxidation in near-bottom water layers ranged from 0.002 to 1.78 microliters/(1 day). Methanogens and methanotrophs were found to play an important role in the carbon cycle through all layers of sediments, particularly in the areas of methane vent and gas-hydrate occurrence.

[The structure and biogeochemical activity of the phototrophic communities from the Bol'sherechenskii alkaline hot spring]. / Struktura i biogeokhimicheskaia aktivnost' fototrofnykh soobshchestv shchelochnogo termal'nogo Bol'sherechenskogo istochnika.

Microbial communities growing in the bed of the alkaline, sulfide hot spring Bol'sherechenskii (the Baikal rift area) were studied over many years (1986-2001). The effluent water temperature ranged from 72 to 74 degrees C, pH was from 9.25 to 9.8, and sulfide content was from 12 to 13.4 mg/ml. Simultaneous effects of several extreme factors restrict the spread of phototrophic microorganisms. Visible microbial fouling appears with a decrease in the temperature to 62 degrees C and in the sulfide content to 5.9 mg/l. Cyanobacteria predominated in all biological zones of the microbial mat. The filamentous cyanobacteria of the genus Phormidium are the major mat-forming organisms, whereas unicellular cyanobacteria and the filamentous green bacterium Chloroflexus aurantiacus are minor components of the phototrophic communities. No cyanobacteria of the species Mastigocladus laminosus, typical of neutral and subacid springs, were identified. Seventeen species of both anoxygenic phototrophic bacteria and cyanobacteria were isolated from the microbial mats, most of which exhibited optimum growth at 20 to 45 degrees C. The anoxygenic phototrophs were neutrophiles with pH optimum at about 7. The cyanobacteria were the most adapted to the alkaline conditions in the spring. Their optimum growth was observed at pH 8.5-9.0. As determined by the in situ radioisotope method, the optimal growth and decomposition rates were observed at 40-32 degrees C, which is 10 to 15 degrees C lower than the same parameter in the sulfide-deficient Octopus Spring (Yellowstone, United States). The maximum chlorophyll a concentration was 555 mg/m2 at 40 degrees C. Total rate of photosynthesis in the mats reached 1.3 g C/m2 per day. The maximum rate of dark fixation of carbon dioxide in the microbial mats was 0.806 g C/m2 per day. The maximum rate of sulfate reduction comprised 0.367 g S/m2 per day at 40 degrees C. The rate of methanogenesis did not exceed 1.188 micrograms C/m2 per day. The role of methanogenesis in the terminal decomposition of the organic matter was insignificant. Methane formation consumed 100 times less organic matter than sulfate reduction.

[Mats of Microcoleus from alkaliphilic and halophilic communities]. / Maty Microcoleus iz alkalofil'nykh i galofil'nykh soobshchestv.

A detailed description of the macrostructure, the ultrastructure, and the species diversity of alkaliphilic mat from Lake Khilganta (Buryatiya) is presented. The structure of this mat was found to be similar to that of halophilic mats from hypersaline lagoons of Lake Sivash (Crimea) that we studied earlier. Microcoleus chthonoplastes was the dominant form of cyanobacteria in both mats (in the alkaliphilic mat, Phormidium molle was another dominant form). Both mats had a pronounced laminated structure. However, unlike halophilic mats with calcium carbonate and gypsum laminas, the alkaliphilic mat contained calcium phosphate laminas instead of gypsum ones. The species diversity of microorganisms in the alkaliphilic mat was at least as rich as that in the halophilic mat; however, in the halophilic mat, the distribution of organisms between layers was more clear-cut. In the alkaliphilic mat, the highest species diversity was observed in the upper mat layers at the boundary between zones of oxygenic and anoxygenic photosynthesis. This fact can be explained by the ephemeral nature of soda lakes.

[Ecophysiological features of mat-forming bacteria Thioploca in bottom sediments of Frolikha Bay, northern Baikal]. / Ekofiziologicheskie osobennosti matobrazuiushche bakterii Thioploca v donnykh osadkakh bukhty Frolikha, severnyi Baikal.

A colorless sulfur bacterium of the genus Thioploca, which forms bacterial mats, was studied in the region of underwater thermal vents (Frolikha Bay, northern Baikal). The organism occurs under microaerobic conditions in top sediment layers, and its biomass can amount to 65 mg of wet weight per 1 kg of silt. Individual filaments of the bacterium penetrate the anaerobic zone to the depth of 19 cm. Thioploca is distributed in a mosaic pattern over the bottom of the bay. Thioploca mats are typically found near vents that discharge low-temperature underground water. In the form of separate filaments, this bacterium is more widely distributed in the top sediment layer, particularly in sediments with a more active sulfate reduction. The bacteria from the deep-water and coastal areas of the bay have different morphology. Cells of Thioploca are able to accumulate nitrate, and the coefficient of nitrate accumulation in wet bacterial mass in relation to the near-bottom water is 1.3 x 10(4), suggesting a similarity of metabolism with seawater species. A more lightweight isotopic composition of nitrogen in cell mass as compared to that of representatives of zoobenthos also indicates an active metabolism of nitrogen, apparently, in the process of nitrogen respiration. Comparison of the composition of stable isotopes of carbon in the biomass of representatives of different trophic levels, including Thioploca, found at a depth of 105 m indicates its planktonic origin, whereas, in the deeper bay region, the biomass of Thioploca incorporates more of the light carbon originating from biogenic methane.

[Effect of bulk discharges of oil products on sludge microbiocenoses of purification works and their function]. / Vliianie zalpovykh vybrosov nefteproduktov na mikrobiotsenozy i funktsional'nuiu aktivnost' biologicheskogo ila ochistnykh sooruzhenii.

The effects of two bulk discharges of oil products on the microbiocenoses and functioning of bioactive sludge in purification works of Ulan-Ude city were investigated. Pollution with oil products exerted both a direct toxic effect on the microorganisms and an indirect effect via food chains. No toxic effects of oil products on sludge nitrifying bacteria were found.

[New genus of budding bacteria Angulomicrobium tetraedrale having radially symmetrical cells]. / Novyi rod pochkuiushchikhsia bakterii Angulomicrobium tetraedrale s radial'noi simmetriei kletki.

The growth cycle and the ultrathin structure were compared in two strains of polygonal bacteria. Due to the presence of radial symmetry, the type of division, the character of metabolism and physiology in common, the polygonal bacteria are classed as a separate genus, Angulomicrobium nov. gen. It would be too early to divide the genus into species. The diagnosis of the new genus and the species A. tetraedrale nov. gen. nov. sp. is presented.