B. B. Namsaraev

Microcoleus Mats from Alkaliphilic and Halophilic Communities

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.

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 enumeration 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 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 changes in salinity 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 low of moderate 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.

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 106–107 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/(m2h); 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.

[The structure and biogeochemical activity of the phototrophic communities from the Bol'sherechenskii alkaline hot spring].

Microbial communities growing in the bed of the alkaline, sulfide hot spring Bolsherechenskii (the Baikal rift area) were studied over many years (1986–2001). The effluent water temperature ranged from 72 to 74°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 mat appears with a decrease in the temperature to 62°C and in 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°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°C, which is 10–15°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°C. The 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°C. The rate of methanogenesis did not exceed 1.188 μg 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.

High abundances of aerobic anoxygenic phototrophs in saline steppe lakes

We studied the distribution of anoxygenic phototrophs in 23 steppe lakes in the Transbaikal region (Russia), in Uzbekistan (Central Asia) and in the Crimean peninsula (Ukraine). The lakes varied in their mineral content and composition (salinities from 0.2 to 300 g L(-1) ). The Transbaikal lakes were alkaline (pH>9), with high amounts of soda. The Uzbek and Crimean lakes were more pH neutral, frequently with high amounts of sulfates. The presence of anoxygenic phototrophs was registered by infrared epifluorescence microscopy, infrared fluorometry and pigment analyses. In mostly shallow, fully oxic lakes, the anoxygenic phototrophs represented 7-65% of the total prokaryotes, with the maxima observed in Transbaikal soda lakes Gorbunka (32%), Khilganta (65%), Zanday (58%) and Zun-Kholvo (46%). Some of the lakes contained over 1 μg bacteriochlorophyll L(-1) . In contrast, only small amounts of anoxygenic phototrophs were present in highly mineralized lakes (>100 g total salts L(-1) ); Borzinskoe, Tsagan-Nur (Transbaikal), Staroe (Crimea) and in the residual part of the south-west Aral Sea (Uzbekistan). The oxic environment and the specific diurnal changes of bacteriochlorophyll concentration observed suggest that the phototrophic community was mostly composed of aerobic anoxygenic phototrophs. The high abundances and bacteriochlorophyll concentrations point to an important role of aerobic anoxygenic phototrophs in the habitats studied.

Microbial communities of the stratified soda Lake Doroninskoe (Transbaikal region)

The physicochemical properties, species composition, and vertical distribution of microorganisms in the water column, shoreline microbial mat, and small shoreline mud volcanoes of the stratified soda Lake Doroninskoe were investigated in September 2007. The lake is located in the Transbaikal region, in the permafrost zone (51°25′N; 112°28′E). The maximal depth of the contemporary lake is about 6 m, the pH value of the water is 9.72, and the water mineralization in the near-bottom horizon is 32.3 g l−1. In summer, the surface oxygen-containing horizon of the water column becomes demineralized to 26.5 g l−1; at a depth of 3.5–4.0 m, an abrupt transition occurs to the aerobic zone containing hydrosulfide (up to 12.56 g l−1). Hydrosulfide was also detected in trace quantities in the upper water horizons. The density stratification of the water column usually ensures stable anaerobic conditions until the freezing period (November and December). The primary production of oxygenic phototrophs reached 176–230 μg l−1. High rates of dark CO2 assimilation (61–240 μg l−1) were detected in the chemocline. Within this zone, an alkaliphilic species of sulfur-oxidizing bacteria of the genus Thioalkalivibrio was detected (104 cells ml−1). Lithoheterotrophic bacteria Halomonas spp., as well as bacteriochlorophyll a-containing aerobic anoxygenic phototrophic bacteria (AAP) Roseinatronobacter sp. capable of thiosulfate oxidation, were isolated from samples collected from the aerobic zone (0–3 m). The water transparency in September was extremely low; therefore, no visible clusters of anoxygenic phototrophic bacteria (APBs) were detected at the boundary of the hydrosulfide layer. However, purple sulfur bacteria which, according to the results of the 16S rRNA gene analysis, belong to the species Thioalkalicoccus limnaeus, Ectothiorhodospira variabilis, “Ect. magna,” and Ect. shaposhnikovii, were isolated from samples of deep silt sediments. Ect. variabilis and Ect. shaposhnikovii were the major APB species in the shoreline algo-bacterial mat. The halotolerant bacterium Ect. shaposhnikovii, purple nonsulfur bacteria of the genus Rhodobacter, and AAP of Roseococcus sp. were isolated from the samples collected from mud volcanoes. All these species are alkaliphiles, moderate halophiles, or halotolerant microorganisms.

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°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°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°C, the filamentous cyanobacterium Phormidium laminosum was predominant; its cell number and biomass concentration comprised 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°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°C. The peak mass of bacterial mats (56.75 g/m2) occurred at a temperature of 65–60°C. The maximum biomass concentration of phototrophs (414.63 × 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°C.

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.

Bacterial Processes of the Methane Cycle in Bottom Sediments of Lake 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 μl CH4/(dm3 day) and oxidized at rates of 0.005–1180 μl 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 μl CH4/(m2 day). Rates of bacterial methane oxidation in near-bottom water layers ranged from 0.002 to 1.78 μl/(l 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 effect of temperature and pH on the growth of aerobic alkalithermophilic bacteria from hot springs in Buryatia

Growth parameters (temperature and pH) were determined for collection cultures of aerobic heterotrophic bacteria. Analysis of the experimental data with the use of the Rosso model made it possible to calculate the extreme values of temperature and pH permissive for culture growth. The cultures examined were subdivided into three groups with respect to their growth temperature and pH. The first group is represented by cultures with minimum, maximum, and optimal growth temperatures of <20, 60–64, and 38–40°C, respectively, and with the optimal growth pH 8.0–8.5. Bacteria of the second group are true alkalithermophilic organisms with a temperature optimum of 45–50°C and a pH optimum of 8.5–9.0. The third group includes a culture of a thermophilic alkalitolerant bacterium.