T. I. Zemskaya

Microbial community structure in methane hydrate-bearing sediments of freshwater Lake Baikal.

Gas hydrates in marine sediments have been known for many years but recently hydrates were found in the sediments of Lake Baikal, the largest freshwater basin in the world. Marine gas hydrates are associated with complex microbial communities involved in methanogenesis, methane oxidation, sulfate reduction and other biotransformations. However, the contribution of microorganisms to the formation of gas hydrates remains poorly understood. We examined the microbial communities in the hydrate-bearing sediments and water column of Lake Baikal using pyrosequencing of 16S rRNA genes. Aerobic methanotrophic bacteria dominated the water sample collected at the lake floor in the hydrate-bearing site. The shallow sediments were dominated by Archaea. Methanogens of the orders Methanomicrobiales and Methanosarcinales were abundant, whereas representatives of archaeal lineages known to perform anaerobic oxidation of methane, as well as sulfate-reducing bacteria, were not found. Affiliation of archaea to methanogenic rather than methane-oxidizing lineages was supported by analysis of the sequences of the methyl coenzyme M reductase gene. The deeper sediments located at 85-90 cm depth close to the hydrate were dominated by Bacteria, mostly assigned to Chloroflexi, candidate division JS1 and Caldiserica. Overall, our results are consistent with the biological origin of methane hydrates in Lake Baikal.

Faunal communities at sites of gas- and oil-bearing fluids in Lake Baikal

Macro- and meiofaunal communities were examined at four geomorphologically distinct sites with different gas- and oil-bearing fluid characteristics in the northern, central and southern basins of Lake Baikal. All sites had elevated concentrations of bicarbonate, nitrate, sulphate and chloride ions in pore fluids, with highest values at the Frolikha vent. Elevated levels of iron ions were found in pore waters of the St. Petersburg methane seep and the Gorevoy Utes oil seep. The chemical composition of pore waters at the Malenky mud volcano was similar to that reported in earlier work. Consistent with published data, the Frolikha vent (northern basin) and the St. Petersburg methane seep (central basin) were characterised by methane of mixed genesis (thermogenic + biogenic), whereas the methane source was mainly thermogenic at the Gorevoy Utes oil seep (central basin) and biogenic at the Malenky mud volcano (southern basin). In contrast to marine seep ecosystems, the macrofauna was dominated only by amphipods, giant planarians and oligochaetes, whereas bivalves were absent; the meiofauna was similar to its marine counterpart, being dominated by nematodes, cyclops, harpacticoids and ostracods. A statistically significant positive relationship was revealed between faunal abundance and the availability of bacterial mats on seep sediments. Moreover, ANOVA tests showed significant increases in both meiozoobenthic and macrozoobenthic densities at “hot spot” vent/seep sites relative to discharge-free reference sites. The isotopic composition of carbon and nitrogen at various trophic levels of these benthic vent/seep communities was found to differ markedly from that reported by earlier studies for the pelagic and other benthic food webs in Lake Baikal. As in marine seeps, the macrofauna had variable isotopic signatures. Light δ13C and δ15N values suggest the utilization of chemosynthetically fixed and/or methane-derived organic matter. By contrast, the heavy δ13C signatures of some mobile amphipods likely reflect consumption of photosynthetically derived carbon. These findings would at least partly explain why Lake Baikal is a notable outlier in global temperature–biodiversity patterns, exhibiting the highest biodiversity of any lake worldwide but at an extremely cold average temperature.

The First Results of an Investigation into the Phylogenetic Diversity of Microorganisms in Southern Baikal Sediments in the Region of Subsurface Discharge of Methane Hydrates

Phylogenetic analysis of the bacterial communities in Lake Baikal bottom sediments in the region of subsurface methane hydrate discharge has been carried out using data on 16S rRNA sequences. The composition of these microbial communities is shown to be different in different horizons. Methanotrophic bacteria are found in the surface layer (0–5 cm), and uncultured bacteria constitute a great portion of this population. In deeper sediment layers (92–96 cm), a change in the microbial community occurs; specifically, a decreased homology with the known sequences is observed. The new sequences form separate clusters on a phylogenetic tree, indicating the possibly endemic nature of the bacteria revealed. Organisms related to the genus Pseudomonas constitute the main portion of the population. An archaea-related sequence was found in a horizon containing gas hydrate crystals (100–128 cm). Uncultured bacteria remain predominant.

Ecophysiological Characteristics of the Mat-forming Bacterium Thioplocain Bottom Sediments of the Frolikha Bay, Northern 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. Thioplocais distributed in a mosaic pattern over the bottom of the bay. Thioplocamats 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 Thioplocaare able to accumulate nitrate, and the coefficient of nitrate accumulation in wet bacterial mass in relation to the near-bottom water is 1.3 × 104, 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 Thioplocaincorporates more of the light carbon originating from biogenic methane.

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.

Activity and structure of the sulfate-reducing bacterial community in the sediments of the southern part of Lake Baikal

The rates of sulfate reduction (SR) and the diversity of sulfate-reducing bacteria (SRB) were studied in the sediments of the Posol’skaya Banka elevation in the southern part of Lake Baikal. SR rates varied from 1.2 to 1641 nmol/(dm3 day), with high rates (>600 nmol/(dm3 day)) observed at both deep-water stations and in subsurface silts. Integral SR rates calculated for the uppermost 50 cm of the sediments were higher for gas-saturated and gas hydrate-bearing sediments than in those with low methane content. Enrichment cultures were obtained in Widdel medium for freshwater SRB. Analysis of the 16S rRNA gene fragments from clone libraries obtained from the enrichments revealed the presence of SRB belonged to the genus Desulfosporosinus, with D. lacus as the most closely related member (capable of sulfate, sulfite, and thiosulfate reduction), as well as members of the order Clostridiales.The rates of sulfate reduction (SR) and the diversity of sulfate-reducing bacteria (SRB) were studied in the sediments of the Posolskaya banka elevation in the southern part of Lake Baikal. SR rates varied from 1.2 to 1641 nmol/(dm3 day), with high rates (> 600 nmol/(dm3 day)) observed at both deep-water stations and in subsurface silts. Integral SR rates calculated for the uppermost 50 cm of the sediments were higher for gas-saturated and gas hydrate-bearing sediments than in those with low methane content. Enrichment SRB cultures were obtained in Widdel medium for freshwater SRB. Analysis of the 16S rRNA gene fragments from clone libraries obtained from the enrichments revealed the presence of SRB belonged to Desulfosporosinus genus, with D. lacus as the most closely related member (capable of sulfate, sulfite, and thiosulfate reduction), as well as members of the order Clostridiales.

[Microbial community of the water column of the Selenga River-Lake Baikal biogeochemical barrier].

The microbial communities of the estuarine zone and the mixing zone of river and lake waters in the Selenga River estuary were studied using the fluorescence in situ hybridization (FISH) method. The microorganisms belonging to the phylogenetic group Gammaproteobacteria were found to predominate in the river estuary, constituting up to 17% of the total bacterial community. Among cultivable microorganisms, organotrophic bacteria were predominant (2040 CFU/ml) in this zone, which results in high rates of microbial production (6.0 μg C/(l day). The microbial community structure changed with distance from the river estuary; representatives of the Alpha-, Beta-, and Gammaproteobacteria were present in equal proportions; psychrotolerant and oligotrophic bacteria were numerous. The rate of heterotrophic carbon dioxide assimilation decreased to 3.8 μg C/(l day). At 5–7 km from the river estuary, where the hydrologic, physical, and chemical conditions are similar to those of lake waters, members of the Betaproteobacteria, which are typical of the open waters of Lake Baikal, are the major representatives of planktonic microorganisms.

Microbial communities of the discharge zone of oil- and gas-bearing fluids in low-mineral Lake Baikal

At the site of natural ingress of oil, microbial diversity in the Central Baikal bottom sediments differing in the chemical composition of pore waters was studied by molecular biological techniques. The sediments saturated with oil and methane were found to contain members of 10 bacterial and 2 archaeal phyla. The oxidized sediment layer contained methanotrophic bacteria belonging to the Alphaproteobacteria, which had a specific structure of the pmoA gene and clustered together with uncultured methanotrophs from cold ecosystems. The upper sediment layer also contained oil-oxidizing bacteria and the alkB genes most closely related to those of Rhodococcus. The microbial community of reduced sediments exhibited lower diversity and was represented mostly by the organisms involved in hydrocarbon biodegradation.

Composition of the Microbial Communities of Bituminous Constructions at Natural Oil Seeps at the Bottom of Lake Baikal

Microbial communities of two bituminous constructions at the bottom of Lake Baikal in the region of natural oil seeps at a depth of 900 m have been investigated. Construction 8 contained biodegraded hydrocarbons, and construction 3, through which oil seeped, contained material that experienced biodegradation to a lesser degree. The composition of the microbial communities was studied by means of pyrosequencing of 16S rRNA gene fragments. Most of the bacterial 16S rRNA gene sequences identified in both bituminous constructions were attributed to proteobacteria, along with which Actinobacteria, Acidobacteria, Bacteroidetes, and TM7 were revealed. About 40% of the bacterial sequences in bituminous construction 3 belonged to representatives of uncultured groups within the classes Alphaproteobacteria and Betaproteobacteria and the phylum Bacteroidetes. The 16S rRNA gene sequences of archaea belonged to aceticlastic and hydrogenotrophic methanogens of the orders Methanosarcinales, Methanomicrobiales, and Methanobacteriales. The 16S rRNA genes of various groups of bacteria carrying out aerobic biodegradation of aromatic compounds and n-alkanes were found; their compositions differed between the constructions. Neither known groups of denitrifying betaproteobacteria nor known groups of sulfate-reducing deltaproteobacteria capable of carrying out anaerobic degradation of n-alkanes were found, which agrees with the low content of nitrate and sulfate in the water. In the anaerobic zone of bituminous constructions, the processes of biodegradation of hydrocarbons are probably carried out in the absence of alternative electron acceptors by the syntrophic community, including deltaproteobacteria of the genus Syntrophus and methanogenic archaea.

Computer system for image analysis of fluorescently stained bacteria

751 The method of fluorescence in situ hybridization (FISH), which enables direct detection of single bacterial cells in natural samples without cultivation, has been widely used over the last few decades for the analysis of structure and dynamics of the microbial community in aquatic ecosystems. This method is based on hybridization of microbial cells with rRNA-specific fluorescent-labeled oligonucleotide probes [1–3]. This method was applied effectively for the analysis of the phylogenetic structure of microbial communities of the water column of two regions of Lake Baikal [4] and for their comparison with marine ecosystems.