N V Pimenov

Winogradsky Institute Of Microbiology , Moscow, Moscow, Russia

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Publications (68)92.96 Total impact

  • Article: The search for sulfate-reducing bacteria in mat samples from the lost city hydrothermal field by molecular cloning
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    ABSTRACT: The work is dedicated to searching for microorganisms of the domain Bacteria capable of dissimilatory sulfate reduction in the samples of microbial mats from a carbonate chimney in the Lost City hydro-thermal field. Cloning of 16S rRNA genes, the universal phylogenetic marker, and dsrAB, the functional marker for sulfate reduction, revealed phylotypes related to spore-forming Desulfotomaculum. No members of the Deltaproteobacteria, comprising the most numerous bacterial group with demonstrated capacity for dissimilatory sulfate reduction, were found. The phylogenetic position of 16S rRNA clones from the mats suggests that this microbial community is a unique consortium, where the energy flow is related to hydrogen of hydrothermal origin, while mass growth of primary produces results from utilization of sulfide formed by sulfate-and sulfur-reducing microorganisms. Key wordsLost City hydrothermal field-microbial diversity-sulfate-reducing bacteria-16S rRNA- dsrAB - Desulfotomaculum
    Microbiology 04/2012; 79(1):96-105. · 3.06 Impact Factor
  • Article: Fractionation of stable carbon isotopes by photoautotrophically growing anoxygenic purple and green sulfur bacteria
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    ABSTRACT: Fractionation of stable carbon isotopes 12C and 13C by three pure cultures of photoautotrophic purple sulfur bacteria (Ectothiorhodospira shaposhnikovii, Lamprocystis purpureus, and Thiocapsa sp.) (PSB) and the green sulfur bacterium Prosthecochloris sp. (GSB) was investigated in 13–15-day experiments. The cultivation was carried out in a luminostat (2000 lx) on mineral media with 1–1.5 g/l NaHCO3 (inoculum) with the subsequent transfer to the medium with up to 10 g/l NaHCO3. For PSB, the difference in the quantitative characteristics of the isotopic composition of suspended carbon (including bacterial cells) and mineral carbon of the medium (Δ13C = δ13Csubstrate − δ13Cbiomass) changed from 15.0 to 34.3‰. For GSB, the range of Δ13C changes was significantly less (18.3–22.7‰). These data suggested the possibility of a pool of soluble mineral carbon in PSB cells. The pool of intracellular mineral carbon was calculated; depending on the PSB species and growth stage, it varied from 0 to 68% of the total cell carbon. The α coefficients reflecting the carbon isotope fractionation by PSB and GBS and calculated from the changes of the bicarbonate carbon isotopic composition in the medium depending on its consumption were 1.029 ± 0.003 and 1.019 ± 0.001, respectively. These α values did not depend on the growth rate. CO2 fixation on ribulose-bisphosphate was shown to be the major factor determining the carbon isotope fractionation by PSB; at the stage of CO2 penetration into the cell, fractionation was insignificant. In GSB, fractionation occurred mostly at CO2 penetration into the cell, while it was insignificant at the stage of carbon dioxide fixation in the reverse TCA cycle. Analysis of the isotopic data of the photosynthesis by PSB and GSB in meromictic lakes also revealed that in PSB-dominated natural communities suspended organic matter was more enriched with light 13C (Δ13C = 23.4−24.6‰) than in the communities with more active GSB (Δ13C = 10.2−14.0‰)
    Microbiology 04/2012; 78(6):757-768. · 3.06 Impact Factor
  • Article: Phylogenetic characterization of endosymbionts of the hydrothermal vent mussel Bathymodiolus azoricus by analysis of the 16S rRNA, cbbL, and pmoA genes
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    ABSTRACT: In order to assess the phylogenetic diversity of the endosymbiotic microbial community of the gills of marine bivalve Bathymodiolus azoricus, total DNA was extracted from the gills. The PCR fragments corresponding to the genes encoding 16S rRNA, ribulose-bisphosphate carboxylase (cbbL), and particulate methane monooxygenase (pmoA) were amplified, cloned, and sequenced. For the 16S rDNA genes, only one phylotype was revealed; it belonged to the cluster of thiotrophic mytilid’s symbionts within the Gammaproteobacteria. For the RuBisCO genes, two phylotypes were found, both belonging to Gammaproteobacteria. One of them was closely related to the previously known mytilid symbiont, the other, to a pogonophore symbiont, presumably a methanotrophic bacterium. One phylotype of particulate methane oxygenase genes was also revealed; this finding indicated the presence of a methanotrophic symbiont. Phylogenetic analysis of the pmoA placed this endosymbiont within the Gammaproteobacteria, in a cluster including the methanotrophic bacterial genus Methylobacter and other methanotrophic Bathymodiolus gill symbionts. These results provide evidence for the existence of two types of endosymbionts (thioautotrophic and methanotrophic) in the gills of B. azoricus and demonstrate that, apart from the phylogenetic analysis of 16S rRNA genes, parallel analysis of functional genes is essential.
    Microbiology 04/2012; 75(6):694-701. · 3.06 Impact Factor
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    Article: Microbial processes at the aerobic-anaerobic interface in the deep-water zone of the black sea
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    ABSTRACT: 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).
    Microbiology 04/2012; 69(4):436-448. · 3.06 Impact Factor
  • Article: Study of the microbial processes in the water column and bottom sediments of the Dolgaya-Vostochnaya Bay (Barents Sea) before construction of the northern tidal power plant
    Microbiology 04/2012; 78(6):798-801. · 3.06 Impact Factor
  • Article: Methane cycle in the Barents Sea
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    ABSTRACT: Biogeochemical cycle of methane in the Barents Sea was studied using isotope geochemistry to determine the rates of microbial methane oxidation. It was established that microbiological processes (glucose consumption, 14CO2 assimilation, sulfate reduction, and slow methane oxidation) in oxidized surface and weakly reduced sediments are marked by only insignificant change in SO42− concentration and absence of notable growth of total alkalinity and N/NH4 downward the sediment core. Microbial methane productivity was 0.111 × 106 mol day−1. Taking into account the volume of water column, microbial methane consumption therein can be as much as 1.8 × 106 mol day−1.
    Lithology and Mineral Resources 04/2012; 43(5):405-428. · 0.38 Impact Factor
  • Article: Investigation of the sulfate-reducing bacterial community in the aerobic water and chemocline zone of the Black Sea by the fish technique
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    ABSTRACT: 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. Keywordsmeromictic basin–Black Sea–aerobic waters–chemocline–sulfate-reducing bacteria–fluorescent in situ hybridization
    Microbiology 04/2012; 80(1):108-116. · 3.06 Impact Factor
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    Article: Methane formation and oxidation in the meromictic oligotrophic Lake Gek-Gel (Azerbaijan)
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    ABSTRACT: The production and oxidation of methane and diversity of culturable aerobic methanotrophic bacteria in the water column and upper sediments of the meromictic oligotrophic Lake Gek-Gel (Azerbaijan) were studied by radioisotope, molecular, and microbiological techniques. The rate of methane oxidation was low in the aerobic mixolimnion, increased in the chemocline, and peaked at the depth where oxygen was detected in the water column. Aerobic methanotrophic bacteria of type II belonging to the genus Methylocystis were identified in enrichment cultures obtained from the chemocline. Methane oxidation in the anaerobic water of the monimolimnion was much more intense than in the aerobic zone. However, below 29–30 m methane concentration increased and reached 68 μM at the bottom. The highest rate of methane oxidation under anaerobic conditions was revealed in the upper layer of bottom sediments. The rate of methane oxidation significantly exceeding that of methane production suggests a deep source of methane in this lake. Key wordsmeromictic oligotrophic Lake Gek-Gel-methanogenesis-methane oxidation-methanotrophic bacteria
    Microbiology 04/2012; 79(2):247-252. · 3.06 Impact Factor
  • Article: Primary production of organic matter and phototrophic communities in the soda lakes of the Kulunda steppe (Altai krai)
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    ABSTRACT: The rates of photosynthesis and dark CO2 fixation were determined in 12 soda lakes of the Kulunda steppe. Characterization of the phototrophic communities was given, and the cell numbers of anoxygenic phototrophic bacteria (APB) were determined. The photosynthetic production in different lakes was substantially different, constituting from 0.01 to 1.32 g C m−2 day−1. The main part of carbon dioxide was assimilated in the process of oxygenic photosynthesis. Anoxygenic photosynthesis was recorded only in 5 of the 12 lakes studied. Its values varied between 0.06 and 0.42 g C m−2 day−1, constituting from 8 to 34% of the total photosynthetic activity. Anoxygenic photosynthesis was revealed in the lakes where the number of APB reached 107–109 CFU cm−3. Dark CO2 fixation constituted 0.01–0.15 g C m−2 day−1. Positive correlation was observed between the primary production value and water alkalinity. No relationship between productivity and water mineralization was revealed in the 30–200 g l−1 range, whereas an increase in salinity above 200 g l−1 suppressed the photosynthetic activity. The mechanisms of influence of the environmental factors on the rate of photosynthesis are discussed.
    Microbiology 04/2012; 78(5):643-649. · 3.06 Impact Factor
  • Article: Radioisotopic tracing of carbon monoxide conversion by anaerobic thermophilic prokaryotes
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    ABSTRACT: The rate of CO conversion by a pure culture of a thermophilic CO-oxidizing, H2-producing bacterium Carboxydocella sp. strain 1503 was determined by the radioisotopic method. The overall daily uptake of 14CO by the bacterium was estimated at 38–56 μmol CO per 1 ml of the culture. A radioisotopic method was developed to separate and quantitatively determine the products of anaerobic CO conversion by microbial communities in hot springs. The new method was first tested on the microbial community from a sample obtained from a hot spring in Kamchatka. The potential rate of CO conversion by the anaerobic microbial community was found to be 40.75 nmol CO/cm3 sediment per day. 85% of the utilized 14CO was oxidized to carbon dioxide; 14.5% was incorporated into dissolved organic matter, including 0.2% that went into volatile fatty acids; 0.5% was used for cell biomass production; and only just over 0.001% was converted to methane.
    Microbiology 04/2012; 76(5):523-529. · 3.06 Impact Factor
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    Article: The effect of microorganisms and seasonal factors on the isotopic composition of particulate organic carbon from the black sea
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    ABSTRACT: The isotopic composition of particulate organic carbon (POC) from the Black Sea deep-water zone was studied during a Russian-Swiss expedition in May 1998. POC from the upper part of the hydrogen sulfide zone (the C-layer) was found to be considerably enriched with the12C isotope, as compared to the POC of the oxycline and anaerobic zone. In the C-layer waters, the concurrent presence of dissolved oxygen and hydrogen sulfide and an increased rate of dark CO2 fixation were recorded, suggesting that the change in the POC isotopic composition occurs at the expense of newly formed isotopically light organic matter of the biomass of autotrophic bacteria involved in the sulfur cycle. In the anaerobic waters below the C-layer, the organic matter of the biomass of autotrophs is consumed by the community of heterotrophic microorganisms; this results in weighting of the POC isotopic composition. Analysis of the data obtained and data available in the literature allows an inference to be made about the considerable seasonable variability of the POC δ13C value, which depends on the ratio of terrigenic and planktonogenic components in the particulate organic matter.
    Microbiology 04/2012; 69(4):449-459. · 3.06 Impact Factor
  • Article: Biological fractionation of stable carbon isotopes at the aerobic/anaerobic water interface of meromictic water bodies
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    ABSTRACT: Mass-spectrometric investigation of carbon isotope composition (δ13C) was carried out for suspended organic matter and dissolved mineral compounds for the water column of some meromictic water bodies differing in salinity and trophic state. As a rule, a more pronounced carbon isotope fractionation (resulting from the metabolism of phytoplankton and anoxygenic phototrophic bacteria) was revealed in the zones of enhanced oxygenic and anoxygenic photosynthesis. Carbon isotope fractionation at the border between oxidized and reduced waters depends both on the activity of microbial communities and on the dominant species of phototrophic microorganisms. Analysis of the distribution profiles of the isotopic composition of suspended organic matter and dissolved mineral carbon revealed active mineralization of the organic matter newly formed via anoxygenic photosynthesis in the monimolimnion by microbial communities, resulting in the release of isotopically light carbon dioxide. Mineral carbon in the anaerobic zones of highly productive meromictic water bodies is therefore enriched with the light 12C isotope.
    Microbiology 11/2008; 77(6):751-759. · 3.06 Impact Factor
  • Article: Anoxygenic phototrophic bacteria of the high-altitude meromictic Lake Gek-Gel, Azerbaijan
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    ABSTRACT: The anoxygenic phototrophic bacterial community of the high-altitude meromictic Lake Gek-Gel (Azerbaijan) was investigated in September 2003. The highest concentration of bacteriochlorophyll e (48 μg/l) was detected at a depth of 30 m; the peak of bacteriochlorophyll a (4.5 μg/l) occurred at 29 m. Phylogenetic analysis revealed that brown-colored green sulfur bacteria Chlorobium phaeobacteroides predominated in the lake. Nonsulfur purple bacteria phylogenetically close to Blastochloris sulfoviridis were found in insignificant amounts; these organisms have not been previously reported in Lake Gek-Gel.
    Microbiology 09/2008; 77(5):602-609. · 3.06 Impact Factor
  • Article: Microbiological and biogeochemical processes in a pockmark of the Gdansk depression, Baltic Sea
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    ABSTRACT: Comprehensive microbiological and biogeochemical investigation of a pockmark within one of the sites of gas-saturated sediments in the Gdansk depression, Baltic Sea was carried out during the 87th voyage of the Professor Shtokman research vessel. Methane content in the near-bottom water and in the underlying sediments indicates stable methane flow from the sediment into the water. In the 10-m water layer above the pockmark, apart from methane anomalies, elevated numbers of microorganisms and enhanced rates of dark CO2 fixation (up to 1.15 µmol C/(l day)) and methane oxidation (up to 2.14 nmol CH4/(l day)) were revealed. Lightened isotopic composition of suspended organic matter also indicates high activity of the near-bottom microbial community. Compared to the background stations, methane content in pockmark sediments increased sharply from the surface to 40–60 ml/dm3 in the 20–30 cm horizon. High rates of bacterial sulfate reduction (SR) were detected throughout the core (0–40 cm); the maximum of 74 µmol S/(dm3 day) was located in subsurface horizons (15–20 cm). The highest rates of anaerobic methane oxidation (AMO), up to 80 µmol/dm3 day), were detected in the same horizon. Good coincidence of the AMO and SR profiles with stoichiometry close to 1: 1 is evidence in favor of a close relation between these processes performed by a consortium of methanotrophic archaea and sulfate-reducing bacteria. Methane isotopic composition in subsurface sediments of the pockmark (from −53.0 to −56.5‰) does not rule out the presence of methane other than the biogenic methane from the deep horizons of the sedimentary cover.
    Microbiology 09/2008; 77(5):579-586. · 3.06 Impact Factor
  • Article: Application of 13C mineral carbon for assessment of the primary production of organic matter in aquatic environments
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    ABSTRACT: The possibility of measuring the rates of light and dark CO2 assimilation using 13C carbonate was demonstrated on Lake Kichier (Marii El). The application of methods utilizing the stable 13C and the radioactive 14C isotopes resulted in comparable values of the rates of light and dark CO2 fixation. Due to its absolute environmental safety, the method with 13C mineral carbon can be recommended as an alternative to radioisotope methods for qualitative measurements of CO2 fixation rates in aquatic ecosystems.
    Microbiology 03/2008; 77(2):224-227. · 3.06 Impact Factor
  • Article: Anoxygenic phototrophic bacterial community of Lake Shira (Khakassia)
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    ABSTRACT: The anoxygenic phototrophic bacterial community of the brackish meromictic Lake Shira (Khakassia) was investigated in August 2001, July 2002, and February–March 2003. In all the periods of investigation, the prevailing microorganisms were purple sulfur bacteria similar to Lamprocystis purpurea in morphology and pigment composition. Their highest number (3 × 105 cells/ml) was recorded in July 2002 at the depth of 15 m. According to 16S rRNA gene analysis, the strain of purple sulfur bacteria isolated in 2001 and designated ShAm01 exhibited 98.6% similarity to the type strain of Thiocapsa roseopersicina and 97.1–94.4% similarity to the type strains of Tca. pendens, Tca. litoralis, and Tca. rosea. The minor microorganisms of the anoxygenic phototrophic bacterial community within the period of investigation were nonsulfur purple bacteria phylogenetically close to Rhodovulum strictum (98.3% similarity, strain ShRb01), Ahrensia kielensis (of 93.9% similarity, strain ShRb02), Rhodomicrobium vannieli (of 99.7% similarity, strain ShRmc01), and green sulfur bacteria, phylogenetically close to Chlorobium limicola (of 98.7% similarity, strain ShCl03).
    Microbiology 07/2007; 76(4):469-479. · 3.06 Impact Factor
  • Article: Seasonal changes in the structure of the anoxygenic photosynthetic bacterial community in Lake Shunet, Khakassia
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    ABSTRACT: 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.
    Microbiology 05/2007; 76(3):368-379. · 3.06 Impact Factor
  • Article: Biogeochemical processes in the Chukchi Sea
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    ABSTRACT: Study of biogeochemical processes in water and sediments of the Chukchi Sea in August 2004 revealed atypical maximums of the concentration of biogenic elements (N, P, and Si) and the rate of microbial sulfate reduction in the surface layer (0–3 cm) of marine sediments. The C: N: P ratio in the organic matter (OM) of this layer does not fit the Redfield-Richards stoichiometric model. Specific features of biogeochemical processes in sea are likely related to the complex dynamics of water, high primary productivity (110–1400 mg C/(m2 day)), low depth of basin (<50 m in 60% of the water area), reduced food chain due to low population of zooplankton, high density of zoobenthos (up to 4230 g m−2), and high activity of microbial processes. Drastic decrease in the concentration of biogenic elements, iodine, total alkalinity (Alk), and population of microorganisms beneath the 0–3 cm layer testify to a large-scale OM decay at the water-seafloor barrier. Our original experimental data support the high annual rate of OM mineralization at the bottom of the Chukchi Sea.
    Lithology and Mineral Resources 01/2007; 42(3):221-239. · 0.38 Impact Factor
  • Article: Organotrophic activity in Kamchatka hot springs with low pH
    Microbiology 02/2006; 75(2):237-239. · 3.06 Impact Factor
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    Article: Sulfate reduction and methanogenesis in the Shira and Shunet meromictic lakes (Khakasia, Russia)
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    ABSTRACT: The biogeochemical and molecular biological study of the chemocline and sediments of saline meromictic lakes Shira and Shunet (Khakasia, Russia) was performed. A marked increase in the rates of sulfate reduction and methanogenesis was revealed at the medium depths of the chemocline. The rates of these processes in the bottom sediments decreased with depth. The numbers of the members of domains Bacteria, Archaea, and of sulfate-reducing bacteria (SRB) were determined by fluorescence in situ hybridization with rRNA specific oligonucleotide probes labeled with horseradish peroxidase and subsequent tyramide signal amplification. In the chemocline, both the total microbial numbers and those of Bacteria were shown to increase with depth. The archaea and SRB were present in almost equal numbers. In the lake sediments, a drastic decrease in microbial numbers with depth was revealed. SRB were found to prevail in the upper sediment layer and archaea in the lower one. This finding correlated with the measured rates of sulfate reduction and methanogenesis.
    Microbiology 01/2006; 75(6):720-726. · 3.06 Impact Factor