Igor I Rusanov

Microbiological Processes of the Carbon and Sulfur Cycles at Cold Methane Seeps of the North Atlantic

Functioning of microbial communities in surface sediments of the Haakon Mosby underwater mud volcano (lat. 72°N) and in gas seepage fields of the Vestnesa Ridge was investigated using Mir-1 and Mir-2 deep-sea submersibles during the 40th voyage of the research vessel Academician Mstislav Keldysh. Large areas of sedimentary deposits of the Haakon Mosby mud volcano (HMMV) and pockmarks of the Vestnesa Ridge (VR) are covered with bacterial mats 0.1 to 0.5 cm thick. The microbial community making up bacterial mats of the HMMV was dominated by large filamentous bacteria with filaments measuring up to 100 μm in length and 2 to 8 μm in width. The occurrence of rosettes allowed the observed filamentous bacteria to be referred to the morphologically similar genera Leucothrix or Thiothrix. Three morphological types of filamentous bacteria were identified in bacterial mats covering VR pockmarks. Filaments of type one are morphologically similar with representatives of the genera Thioploca or Desmanthos. Type two filaments had numerous inclusions of sulfur and resembled representatives of the genus Thiothrix. The third morphological type was constituted by single filaments made up of tightly connected disk-like cells and can be assigned to the genus Beggiatoa. The rates of methane oxidation (up to 1570 μl C/(dm3 day)) and sulfate reduction (up to 17 mg S/(dm3day)) measured in the surface sediments of HMMV and VR were close to the maximum rates of these processes observed in heavily polluted regions of the northwestern shelf of the Black Sea. High rates of microbiological processes correlated with the high number of bacteria. The rate of methane production in sediments studied was notably lower and ranged from 0.1 to 3.5 μ CH4/(dm3 day). Large areas of the HMMV caldera were populated by pogonophoras, represented by the two species Sclerolinum sp. and Oligobrachia sp. The mass development of Sclerolinum sp. in the HMMV caldera was by the activity of aerobic methane-oxidizing bacteria localized inside the cells of these animals. Bacterial cells were also found in the trophosome tissue of Oligobrachia sp., but in cells of these bacteria, we did not observe the membrane structures typical of methanotrophs. The localization pattern of pogonophoras on the surface of reduced sediments suggests that the predominant bacteria in Oligobrachia tissues are sulfur-oxidizing endosymbionts.

Sulfidogenesis under extremely haloalkaline conditions in soda lakes of Kulunda Steppe (Altai, Russia).

Sulfidogenic activity (SA) in anoxic sediments of several soda lakes with variable salinity in south Kulunda Steppe (Altai, Russia) has been investigated. The study included in situ measurements of sulfate reduction rates and laboratory experiments with sediment slurries in which sulfate, thiosulfate or elemental sulfur were used as electron acceptors. Despite the extreme conditions (high salt concentrations and high pH), the SA values were relatively high (ranging from 0.02 to 1.20 micromol HS(-) cm(-3) h(-1)), and only hampered under salt-saturated conditions. The highest SA was observed with elemental sulfur, followed by thiosulfate, while the lowest SA was determined in the presence of sulfate. Of all the electron donors tested, the addition of formate resulted in the highest SA with all three sulfur electron acceptors. Surprisingly, hydrogen as an electron donor had very little effect. Acetate was utilized as an electron donor only under sulfur-reducing conditions. Indigenous populations of sulfidogens in soda lake sediments showed an obligately alkaliphilic pH response of SA, showing a pattern that corresponded well to the in situ pH conditions. Sulfate reduction was much more susceptible to salt inhibition than thiosulfate and sulfur reduction. Microbiological investigations indicated that sulfate-reducing bacteria belonging to the orders Desulfovibrionales and Desulfobacterales could very likely be responsible for the SA with sulfate and thiosulfate as electron acceptors at moderate salt concentrations. Sulfur reduction at moderate salinity was carried out by a specialized group of haloalkaliphilic sulfur-reducing bacteria that utilize volatile fatty acids. In saturated soda brine, extremely natronophilic representatives of the order Halanaerobiales were responsible for the sulfur-dependent respiration.

Microbial Processes of the Carbon and Sulfur Cycles in Lake Shira (Khakasia)

Microbiological and biogeochemical studies of the meromictic saline Lake Shira (Khakasia) were conducted. In the upper part of the hydrogen-sulfide zone, at a depth of 13.5–14 m, there was a pale pink layer of water due to the development of purple bacteria (6 × 105 cells/ml), which were assigned by their morphological and spectral characteristics toLamprocystis purpurea (formerly Amoebobacter purpureus). In August, the production of organic matter (OM) in Lake Shira was estimated to be 943 mg C/(m2day). The contribution of anoxygenic photosynthesis was insignificant (about 7% of the total OM production). The share of bacterial chemosynthesis was still less (no more than 2%). In the anaerobic zone, the community of sulfate-reducing bacteria played a decisive role in the terminal decomposition of OM. The maximal rates of sulfate reduction were observed in the near-bottom water (114 μg S/(l day)) and in the surface layer of bottom sediments (901 μg S/(dm3 day)). The daily expenditure of Corg for sulfate reduction was 73% of Corg formed daily in the processes of oxygenic and anoxygenic photosynthesis and bacterial chemosynthesis. The profile of methane distribution in the water column and bottom sediments was typical of meromictic reservoirs. The methane content in the water column increased beginning with the thermocline (7–8 m) and reached maximum values in the near-bottom water (17 μl/l). In bottom sediments, the greatest methane concentrations (57 μl/l) were observed in the surface layer (0–3 cm). The integral rate of methane formation in the water column and bottom sediments was almost an order of magnitude higher than the rate of its oxidation by aerobic and anaerobic methanotrophic microorganisms.

Microbial processes of the carbon and sulfur cycles in the White Sea

The present paper contains the results of our microbiological and biogeochemical investigations carried out during a series of expeditions to the White Sea in 2002–2006. The studies were conducted in the open part of the White Sea, as well as in the Onega, Dvina, and Kandalaksha bays. In August 2006, the photosynthetic productivity in the surface water layer was low (47–145 mg C m−2 day−1). Quantitative characteristics of microbial numbers and activity of the the key microbial processes occurring in the water column of the White Sea were explored. Over the 5-year period of observations, the total number of bacterial cells in the surface layer of the water column varied from 50 to 600 thousand cells ml−1. In August 2006, bacterioplankton production (BP) was estimated to be 0.26–3.3 μg C l−1 day−1; the P/B coefficient varied from 0.22 to 0.93. The suspended organic matter had a lighter isotope composition (from −28.0 to −30.5‰) due to the predominance of terrigenous organic matter delivered by the Northern Dvina waters. The interseasonal and interannual variation coefficients for phytoplankton production and BP numbers are compared. The bacterioplankton community of the White Sea’s deep water was found to be more stable than that of the surface layer. In the surface layer of bottom sediments, methane concentration was 0.2–5.2 μl dm−3; the rate of bacterial sulfate reduction was 18–260 μg S dm−3 day−1; and the rates of methane production and oxidation were 24–123 and 6–13 nl CH4 dm−3 day−1, respectively. We demonstrated that the rates of microbial processes of the carbon and sulfur cycles occurring in the sediments of the White Sea basin were low.

[Seasonal changes in the structure of the anoxygenic photosynthetic bacterial community in Lake Shunet, Khakassia].

Seasonal studies of the anoxygenic phototrophic bacterial community of the water column of the saline eutrophic meromictic Lake Shunet (Khakassia) were performed in 2002 (June) and 2003 (February–March and August). From the redox zone down, the lake water was of dark green color. Green sulfur bacteria predominated in every season. The maximum number of green sulfur bacteria was 107 cells/ml in summer and 106 cells/ml in winter. A multi-syringe stratification sampler was applied for the study of the fine vertical distribution of phototrophs in August 2003; the sampling was performed every 5 cm. A 5-cm-thick pink-colored water layer inhabited by purple sulfur bacteria was shown to be located above the layer of green bacteria. The species composition and ratio of purple bacterial species depended on the sampling depth and on the season. In summer, the number of purple sulfur bacteria in the layer of pink water was 1.6 × 108 cells/ml. Their number in winter was 3 × 105 cells/ml. In the upper oxygen-containing layer of the chemocline the cells of purple nonsulfur bacteria were detected in summer. The maximum number of nonsulfur purple bacteria, 5 × 102 cells/ml, was recorded in August 2003. According to the results of the phylogenetic analysis of pure cultures of the isolated phototrophic bacteria, which were based on 16S rDNA sequencing, green sulfur bacteria were close to Prosthecochloris vibrioformis, purple sulfur bacteria, to Thiocapsa and Halochromatium species, and purple nonsulfur bacteria, to Rhodovulum euryhalinum and Pinkicyclus mahoneyensis.

Microbial processes at the aerobic-anaerobic interface in the deep-water zone of the black sea

Chemical and key microbiological processes (assimilation of carbon dioxide, oxidation and formation of methane, and sulfate reduction) occurring at the aerobic-anaerobic interface in the deep-water zone of the Black Sea were investigated. Measurements were taken at depths from 90 to 300 m at intervals of 5–10 m. The integral rate of the dark assimilation of carbon dioxide varied from 120 to 207 mg C/(m2 day) with a maximum at the boundary of cyclonic currents. The organic matter (OM) formed from methane comprised less than 5% of the OM formed from carbon dioxide. A comparison between the rates of methane oxidation and methane production suggests that methane that is oxidized at depths from 100 to 300 m was formed in deeper water horizons. The maximum rate of sulfate reduction (1230 mg S/(m2 day)) was observed in the western halistatic region, and the minimum rate (490 mg S/(m2 day)), in the eastern halistatic region. The average rate of hydrogen sulfide production measured at three deep-sea stations amounted to 755 mg S/(m2 day), or 276 g S/(m2 year).

Microbial processes of the carbon and sulfur cycles in the Kara Sea

The results of microbiological, biogeochemical, and isotope geochemical studies in the Kara Sea are described. The samples for these studies were obtained during the 54th voyage of the research vessel Akademik Mstislav Keldysh in September 2007. The studied area covered the northern, central, and southwestern parts of the Kara Sea and the Gulf of Ob. The quantitative characteristics of the total bacterial population and the activity of the microbial processes in the water column and bottom sediments were obtained. The total population of the bacterioplankton (BP) varied from 250000 cells/ml in the northern water area to 3000000 cells/ml in the Gulf of Ob. The BP population depended on the content of the water suspension. The net BP production was minimal in the central water area, amounting to 0.15–0.2 μg C/(l day), and maximal (0.5–0.75 μg C/(l day)) in the Gulf of Ob. The organic material at the majority of the stations in the Ob transect predominantly contained light carbon isotopes (−28.0 to −30.18‰) of terrigenous origin. The methane content in the surface water layer varied from 0.18 to 2.0 μl CH4/l, and the methane oxidation rate changed in the range of 0.1–100 nl CH4/(l day). The methane concentration in the upper sediment layer varied from 30 to 300 μl CH4/dm3; the rate of the methanogenesis was 44 to 500 nl CH4/(dm3 day) and that of the methane oxidation, 30 to 2000 nl CH4/(dm3 day). The rate of the sulfate reduction varied from 4 to 184 μg S/(dm3 day).

Microbiological and isotopic-geochemical investigations of meromictic lakes in Khakasia in winter

Microbiological and isotopic-geochemical investigations of the brackish meromictic lakes Shira and Shunet were performed in the steppe region of Khakasia in winter. Measurements made with a submersed sensor demonstrated that one-meter ice transmits light in a quantity sufficient for oxygenic and anoxygenic photosynthesis. As in the summer season, in the community of phototrophic bacteria found in Lake Shira, the purple sulfur bacteria Amoebobacter purpureus dominated, whereas, in Lake Shunet, the green sulfur bacteria Pelodictyon luteolum were predominant. Photosynthetic production, measured using the radioisotopic method, was several times lower than that in summer. The rates of sulfate reduction and production and oxidation of methane in the water column and bottom sediments were also lower than those recorded in summer. The process of anaerobic methane oxidation in the sediments was an exception, being more intense in winter than in summer. The data from radioisotopic measurements of the rates of microbial processes correlate well with the results of determination of the isotopic composition of organic and mineral carbon (δ13C) and hydrogen sulfide and sulfate (δ34S) and suggest considerable seasonal variations in the activity of the microbial community in the water bodies investigated.

Investigation of the sulfate-reducing bacterial community in the aerobic water and chemocline zone of the Black Sea by the fish technique

Fluorescent in situ hybridization (FISH) was used to analyze the abundance and phylogenetic composition of sulfate-reducing bacteria in the aerobic waters and in the oxic/anoxic transitional zone (chemocline) of the Black Sea, where biogenic formation of reduced sulfur compounds was detected by radioisotope techniques. Numerous sulfate-reducing bacteria of the genera Desulfotomaculum (30.5% of detected bacterial cells), Desulfovibrio (29.6%), and Desulfobacter (6.7%) were revealed in the aerobic zone at a depth of 30 m, while Desulfomicrobium-related bacteria (33.5%) were prevalent in the upper chemocline waters at 150-m depth. Active cells of sulfate-reducing bacteria were much more abundant in the samples collected in summer than in the winter samples from the deep-sea zone. The presence of physiologically active sulfate reducers in oxic and chemocline waters of the Black Sea correlates with the hydrochemical data on the presence of reduced sulfur compounds in the aerobic water column.

The Biogeochemical Cycle of Methane in the Coastal Zone and Littoral of the Kandalaksha Bay of the White Sea

Microbiological and biogeochemical investigations of the processes of methane production (MP) and methane oxidation (MO) in the coastal waters and littoral of the Kandalaksha Bay of the White Sea were carried out. The studies were conducted in the coastal zones and in the water areas of the Kandalaksha Preserve, Moscow State University White Sea Biological Station, and the Zoological Institute (RAS) biological station in August 1999, 2000, and 2001 and in March 2001. The rate of CO2 assimilation in the shallow and littoral sediments was 35–27 800 μg C/(dm3 day) in summer and 32.8–88.9 μg C/(dm3 day) in winter. The maximal rates of MP were observed in the littoral sediments in the zone of macrophyte decomposition, in local depressions, and in the estuary of a freshwater creek (up to 113 μl/(dm3 day)). The maximal level of MO was observed in the shallow estuarine sediments (up to 2450 μl/(dm3 day)). During the winter season, at the temperature of –0.5 to 0.5°C, the MP rate in the littoral sediments was 0.02–0.3 μl/(dm3 day), while the MO rate was 0.06–0.7 μl/(dm3 day). The isotopic data obtained indicate that the Corg of the mats and of the upper sediment layers is enriched with the heavy 13C isotope by 1–4‰ as compared to the Corg of the suspension. A striking difference was found between the levels of methane emission by the typical littoral microlandscapes. In fine sediments, the average emission was 675 μl CH4/(m2 day); in stormy discharge stretch sediments, it was 1670 μl CH4/(m2 day); and under stones and in silted pits, 1370 μl CH4/(m2day). The calculation, performed with consideration of the microlandscape areas with a high production, allowed the CH4 production of 1 km2 of the littoral to be estimated as 192–300 l CH4/(km2 day).