G B Slobodkina

Winogradsky Institute Of Microbiology, Moskva, Moscow, Russia

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Publications (20)35.05 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: A novel hyperthermophilic, anaerobic, archaeon was isolated from a terrestrial hot spring at Uzon Caldera, Kronotsky Nature Reserve, Kamchatka, Russia. The new isolate, strain 1860T, grew optimally at 90-95oC and pH 6.0-7.0. The cells were non-motile straight rods 1.5-5.0 µm in length covered with surface-layer lattice. Strain 1860T utilized complex proteinaceous compounds as electron donors and ferrihydrite, Fe(III) citrate, nitrate, thiosulfate, selenite, selenate and arsenate as electron acceptors for growth. The sequence of the 16S rRNA gene of strain 1860T had 97.9-98.7% similarity with members of the genus Pyrobaculum. On the basis of its physiological properties and phylogenetic analyses including in silico genome-to genome hybridization, the isolate is considered to represent a novel species, for which the name Pyrobaculum ferrireducens sp.nov. is proposed. The type strain is 1860T (=DSM 28942T = VKM B-2856T).
    International journal of systematic and evolutionary microbiology. 12/2014;
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    ABSTRACT: Two novel strains of thermophilic planctomycetes were recovered from terrestrial and subterranean habitats. Strain R1Twas isolated from a hot spring (Kunashir Island, Russia) and the strain SBP2T was isolated from a deep gold mine (South Africa). Both isolates grew in the temperature range from 30oC to 60oC and pH range 5.0-8.0. The isolate R1T grew optimally at 60oC and pH 6.0 - 6.5; for SBP2T optimal conditions were at 52oC and pH 7.5-8.0. Both strains were capable of anaerobic respiration with nitrate and nitrite as electron acceptor as well as of microaerobic growth. They also could grow by fermentation of mono-, di- and polysaccharides. Based on the phylogenetic position and phenotypic features we suggest that new isolates represent two novel species belonging to the new genus of the order Planctomycetales for which the names Thermogutta terrifontis gen. nov., sp. nov. and Thermogutta hypogea sp. nov. are proposed. The type strain of Thermogutta terrifontis, the type species of the genus, is R1T (= DSM 26237T, = VKM B-2805T), the type strain of the Thermogutta hypogea is SBP2T (= JCM19991T=. VKM B-2782T).
    International journal of systematic and evolutionary microbiology. 12/2014;
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    ABSTRACT: A novel moderately thermophilic bacterium, strain STGH(T), was isolated from Severo-Stavropolskoye underground gas storage (Russia). Cells of strain STGH(T) were spore-forming motile straight rods 0.3 μm in diameter and 2.0-4.0 μm in length having a Gram-positive cell wall structure. The temperature range for growth was 36-65 °C, with an optimum at 50-52 °C. The pH range for growth was 5.5-8.0, with an optimum at pH 7.0-7.5. Growth of strain STGH(T) was observed at NaCl concentrations ranging from 0 to 4.0 % (w/v) with an optimum at 1.0 % (w/v). Strain STGH(T) grew anaerobically by reduction of nitrate, thiosulfate, S(0) and AQDS using a number of complex proteinaceous compounds, organic acids and carbohydrates as electron donors. Nitrate was reduced to nitrite; thiosulfate and sulfur were reduced to sulfide. It also was able to ferment pyruvate, glucose, fructose, and maltose. The strain STGH(T) did not grow under aerobic conditions during incubation with atmospheric concentration of oxygen but was able to microaerobic growth (up to 10 % of oxygen in gas phase). The G+C content of DNA of strain STGH(T) was 34.8 mol%. 16S rRNA gene sequence analysis revealed that the isolated organism belongs to the class Bacilli. We propose to assign strain STGH(T) to a new species of a novel genus Tepidibacillus fermentans gen. nov., sp.nov. The type strain is STGH(T) (=DSM 23802(T), =VKM B-2671(T)).
    Extremophiles 07/2013; · 2.20 Impact Factor
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    ABSTRACT: A thermophilic, anaerobic, chemolithoautotrophic bacterium (strain S69T) was isolated from a deep-sea hydrothermal vent chimney located on the Eastern Lau Spreading Center and Valu Fa Ridge, Pacific Ocean at a depth of 1910 m using anoxic medium with elemental sulfur as the only energy source. Cells of strain S69T were Gram-negative short rods, 0.4 to 0.6 µm in diameter and 1.0 to 2.5 µm in length, motile with a single polar flagellum. The temperature range for growth was 28-70°C, with an optimum at 61°C. The pH range for growth was 5.6-7.9, with an optimum at 6.8. Growth of strain S69T was observed at NaCl concentrations ranging from 0.9 to 5.0% with an optimum at 1.8-2.7 (w/v). Strain S69T grew anaerobically with elemental sulfur as an energy source and bicarbonate/CO(2) as a carbon source. Elemental sulfur was disproportionated to sulfide and sulfate. The growth was enhanced in the presence of poorly crystalline Fe(III) oxide (ferrihydrite) as a sulfide-scavenging agent. Strain S69T was also able to grow by disproportionation of thiosulfate and sulfite. Sulfate was not used as an electron acceptor either with H(2) or organic electron donors. Analysis of the 16S rRNA gene sequence revealed that the isolate formed a distinct phylogenetic branch within the Deltaproteobacteria. On the basis of its physiological properties and results of phylogenetic analyses, it is proposed that the new isolate represents the species of a novel genus, Dissulfuribacter thermophilus gen. nov., sp. nov. with the type strain S69T (=DSM 25762T =VKM B-2760T).
    INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY 09/2012; · 2.11 Impact Factor
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    ABSTRACT: Strain 1860, a novel member of the genus Pyrobaculum, is a hyperthermophilic organotrophic crenarchaeon growing anaerobically with various electron acceptors. The complete genome sequence reveals genes for several membrane-bound oxidoreductases, the Embden-Meyerhof and Entner-Doudoroff pathways for glucose metabolism, the tricarboxylic acid cycle, the glyoxylate cycle, and the dicarboxylate/4-hydroxybutyrate cycle.
    Journal of bacteriology 02/2012; 194(3):727-8. · 3.94 Impact Factor
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    ABSTRACT: An extremely thermophilic, anaerobic, chemolithoautotrophic bacterium (strain S95T) was isolated from a deep-sea hydrothermal vent chimney located on the Eastern Lau Spreading Center, Pacific Ocean at a depth of 1910 m. Cells of strain S95T were oval to short Gram-negative rods, 0.5 to 0.6 µm in diameter and 1.0 to 1.5 µm in length, growing singly or in pairs. Cells were motile with a single polar flagellum. The temperature range for growth was 50-92°C, with an optimum at 74°C. The pH range for growth was 5.5-8.0, with an optimum at 7.0. Growth of strain S95T was observed at NaCl concentrations ranging from 1.5 to 3.5% (w/v). Strain S95T grew anaerobically with elemental sulfur as an energy source and bicarbonate/CO2 as a carbon source. Elemental sulfur was disproportionated to sulfide and sulfate. The growth was enhanced in the presence of poorly crystalline Fe(III) oxide (ferrihydrite) as a sulfide-scavenging agent. Strain S95T was also able to grow by disproportionation of thiosulfate and sulfite. Sulfate was not used as an electron acceptor. Analysis of the 16S rRNA gene sequence revealed that the isolate belongs to the phylum Thermodesulfobacteria. On the basis of its physiological properties and results of phylogenetic analyses, it is proposed that the new isolate represents the sole species of a novel genus, Thermosulfurimonas dismutans gen. nov., sp. nov. with the type strain S95T (=DSM 24515T =VKM B-2683T). Thermosulfurimonas dismutans is a first described thermophilic microorganism that disproportionates elemental sulfur.
    INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY 12/2011; · 2.11 Impact Factor
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    ABSTRACT: A moderately thermophilic, anaerobic, dissimilatory iron(III)-reducing bacterium (strain S3R1(T)) was isolated from a deep-sea hydrothermal vent chimney located on the Eastern Lau Spreading Centre in the Pacific Ocean at a depth of about 2150 m. Cells of strain S3R1(T) were ovals to short rods with a single polar flagellum, Gram-stain-negative, 0.5-0.6 µm in diameter and 0.8-1.3 µm long, growing singly or in pairs. The temperature range for growth was 36-62 °C, with an optimum at 50 °C. The pH range for growth was 5.5-7.5, with an optimum at pH 6.5. Growth of strain S3R1(T) was observed at NaCl concentrations ranging from 1.0 to 5.0 % (w/v), with an optimum at 2.0-2.5 % (w/v). The isolate used acetate, fumarate, malate, maleinate, succinate, propanol, palmitate, stearate, peptone and yeast extract as electron donors for growth and iron(III) reduction. All electron donors were oxidized completely to CO(2) and H(2)O. Iron(III) (in the form of ferrihydrite, ferric citrate or ferric nitrilotriacetate) and elemental sulfur (S(0)) were the electron acceptors that supported growth. The DNA G+C content was 64.4 mol%. Results of 16S rRNA gene sequence analysis showed that the novel bacterium was related to representatives of the orders Desulfuromonadales and Syntrophobacterales with 84-86 % sequence similarity and formed a distinct phylogenetic branch in the Deltaproteobacteria. On the basis of its physiological properties and results of phylogenetic analyses, it is proposed that the new isolate represents the sole species of a novel genus, Deferrisoma camini gen. nov., sp. nov. The type strain of Deferrisoma camini is S3R1(T) ( = DSM 24185(T)  = VKM B-2672(T)).
    INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY 12/2011; 62(Pt 10):2463-8. · 2.11 Impact Factor
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    ABSTRACT: A novel anaerobic, moderately thermophilic bacterium (strain SGL43(T)) was isolated from Severo-Stavropolskoye underground gas storage reservoir (Russia). Cells of strain SGL43(T) were motile straight rods, 0.4 µm in diameter and 2.0-3.0 µm in length. The temperature range for growth was 28-65 °C, with optimum growth at 50 °C. The pH range for growth was 5.5-8.0, with optimum growth at pH 7.0-7.5. Growth of strain SGL43(T) was observed at NaCl concentrations of 0-4.0% (w/v) with optimum growth at 1.0% (w/v) NaCl. Substrates utilized by strain SGL43(T) included peptone, yeast extract, glucose, fructose, maltose, galactose, pyruvate and citrate. Products of glucose or citrate fermentation were acetate, hydrogen and CO(2). Thiosulfate was reduced to sulfide. The DNA G+C content of strain SGL43(T) was 43.1 mol%. 16S rRNA gene sequence analysis revealed that strain SGL43(T) belongs to the order Thermoanaerobacterales (phylum 'Firmicutes'). The closest relative of strain SGL43(T) was Thermoanaerobacterium saccharolyticum (86.2% 16S rRNA gene sequence similarity with the type strain). Based on the data presented here, strain SGL43(T) is considered to represent a novel species of a new genus, for which the name Caloribacterium cisternae gen. nov., sp. nov. is proposed. The type strain of Caloribacterium cisternae, the type species of the genus, is SGL43(T) (=DSM 23830(T)=VKM B-2670(T)).
    INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY 08/2011; 62(Pt 7):1543-7. · 2.11 Impact Factor
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    ABSTRACT: A thermophilic, anaerobic, dissimilatory Mn(IV)- and Fe(III)-reducing bacterium (strain SLM 61T) was isolated from a terrestrial hot spring on the Kamchatka peninsula. The cells were straight rods, 0.5-0.6 µm in diameter and 1.0-6.0 µm long, and exhibited tumbling motility by means of peritrichous flagellation. The strain grew at 26-70 °C, with an optimum at 58-60 °C, and at pH 5.5-8.0, with an optimum at pH 6.5. Growth of SLM 61T was observed at 0-2.0 % (w/v) NaCl, with an optimum at 0.5 % (w/v). The generation time under optimal growth conditions was 40 min. Strain SLM 61T grew and reduced Mn(IV), Fe(III) or nitrate with a number of organic acids and complex proteinaceous compounds as electron donors. It was capable of chemolithoautotrophic growth using molecular hydrogen as an electron donor, Fe(III) but not Mn(IV) or nitrate as an electron acceptor and CO2 as a carbon source. It also was able to ferment pyruvate, yeast extract, glucose, fructose, sucrose and maltose. The G+C content of DNA of strain SLM 61T was 50.9 mol%. 16S rRNA gene sequence analysis revealed that the closest relative of the isolated organism was Carboxydocella thermautotrophica 41T (96.9 % similarity). On the basis of its physiological properties and phylogenetic analyses, the isolate is considered to represent a novel species, for which the name Carboxydocella manganica sp. nov. is proposed. The type strain is SLM 61T (=DSM 23132T=VKM B-2609T). C. manganica is the first described representative of the genus Carboxydocella that possesses the ability to reduce metals and does not utilize CO.
    INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY 06/2011; 62(Pt 4):890-4. · 2.11 Impact Factor
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    ABSTRACT: An anaerobic, thermophilic, spore-forming bacterium (strain 64-FGQ(T)) was isolated from a terrestrial hydrothermal spring from the Kamchatka peninsula, Russia. This strain utilized lactate as an electron donor, insoluble poorly crystalline Fe(III) oxide incorporated into alginate beads as a potential electron acceptor and 9,10-anthraquinone-2,6-disulfonate (AQDS) as an electron-shuttling compound. Vegetative cells of strain 64-FGQ(T) were Gram-stain-positive, peritrichously flagellated, motile, straight rods, 0.3-0.5 µm in diameter and 2.0-5.0 µm long, growing singly or forming short chains. Cells formed round refractive endospores in terminal swollen sporangia. The temperature range for growth was 46-70 °C, with an optimum at 65 °C. The pH range for growth was 5.5-8.5, with an optimum at pH 7.0. The substrates utilized by strain 64-FGQ(T) in the presence of AQDS as an electron acceptor included lactate, malate, succinate, glycerol and yeast extract. The strain fermented galactose, fructose, maltose, sucrose, pyruvate and peptone. Strain 64-FGQ(T) used AQDS, humic acid, thiosulfate, nitrate and perchlorate as electron acceptors for growth. Fe(III) was not directly reduced, but strain 64-FGQ(T) was able to grow and reduce Fe(III) oxide in the presence of small amounts of AQDS or humic acid as electron-shuttling compounds. The G+C content of the DNA of strain 64-FGQ(T) was 51 mol%. 16S rRNA gene sequence analysis placed the isolate in the genus Moorella, with the type strain of Moorella glycerini as its closest relative (97.2% similarity). Based on phylogenetic analysis and physiological characteristics, strain 64-FGQ(T) is considered to represent a novel species of the genus Moorella, for which the name Moorella humiferrea sp. nov. is proposed; the type strain is 64-FGQ(T) (=DSM 23265(T)=VKM B-2603(T)).
    INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY 04/2011; 62(Pt 3):613-7. · 2.11 Impact Factor
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    ABSTRACT: Thirty enrichment cultures of thermophilic microorganisms were obtained from Kamchatka terrestrial hydrotherms that reduced insoluble poorly crystalline Fe(III) oxide (ferrihydrite) with and without direct contact between the cells and the mineral. Restricted access to the Fe(III) mineral was achieved by incorporation of ferrihydrite into alginate beads. According to phylogenetic analysis of 22 enrichment cultures by denaturing gradient gel electrophoresis of 16S rRNA gene fragments, Firmicutes were predominant among bacteria in all the variants. Microorganisms of the phylogenetic types Aquificae, Bacteroidetes, Nitrospirae, Planctomycetes, Spirochaetes, Synergistetes, and Thermotogae were also revealed. The archaea revealed belonged to the genera Desulfurococcus, Pyrobaculum, and Thermofilum. In the case of free access to ferrihydrite, most of the phylotypes belonged to genera known for Fe(III) reduction. In the absence of direct contact with the mineral, together with known iron reducers, organisms for which ability to reduce Fe(III) was unknown were detected. Members of the genera Carboxydothermus, Thermoanaerobacter, and Thermotoga were detected most often both in the presence and absence of contact with ferrihydrite. These organisms probably possess efficient mechanisms for Fe(III) reduction within the experimental temperature range. Microbial phylogenetic diversity was higher when acetate, rather than lactate, was used as a potential electron donor. Key wordsthermophilic microorganisms-microbial Fe(III) reduction-ferrihydrite-insoluble electron acceptors-phylogenetic analysis
    Microbiology 01/2010; 79(5):663-671. · 0.65 Impact Factor
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    ABSTRACT: A hyperthermophilic, anaerobic, dissimilatory Fe(III)-reducing, facultatively chemolithoautotrophic archaeon (strain SBH6(T)) was isolated from a hydrothermal sample collected from the deepest of the known World Ocean hydrothermal fields, Ashadze field (1 degrees 58' 21'' N 4 degrees 51' 47'' W) on the Mid-Atlantic Ridge, at a depth of 4100 m. The strain was enriched using acetate as the electron donor and Fe(III) oxide as the electron acceptor. Cells of strain SBH6(T) were irregular cocci, 0.3-0.5 mum in diameter. The temperature range for growth was 50-85 degrees C, with an optimum at 81 degrees C. The pH range for growth was 5.0-7.5, with an optimum at pH 6.8. Growth of SBH6(T) was observed at NaCl concentrations ranging from 1 to 6 % (w/v) with an optimum at 2.5 % (w/v). The isolate utilized acetate, formate, pyruvate, fumarate, malate, propionate, butyrate, succinate, glycerol, stearate, palmitate, peptone and yeast extract as electron donors for Fe(III) reduction. It was also capable of growth with H(2) as the sole electron donor, CO(2) as a carbon source and Fe(III) as an electron acceptor without the need for organic substances. Fe(III) [in the form of poorly crystalline Fe(III) oxide or Fe(III) citrate] was the only electron acceptor that supported growth. 16S rRNA gene sequence analysis revealed that the closest relative of the isolated organism was Geoglobus ahangari 234(T) (97.0 %). On the basis of its physiological properties and phylogenetic analyses, the isolate is considered to represent a novel species, for which the name Geoglobus acetivorans sp. nov. is proposed. The type strain is SBH6(T) (=DSM 21716(T) =VKM B-2522(T)).
    International journal of systematic and evolutionary microbiology 08/2009; 59(Pt 11):2880-3. · 2.11 Impact Factor
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    ABSTRACT: A thermophilic, anaerobic, chemolithoautotrophic bacterium (designated strain SL50(T)) was isolated from a hydrothermal sample collected at the Mid-Atlantic Ridge from the deepest of the known World ocean hydrothermal fields, Ashadze field (1 degrees 58' 21'' N 4 degrees 51' 47'' W) at a depth of 4100 m. Cells of strain SL50(T) were motile, straight to bent rods with one polar flagellum, 0.5-0.6 mum in width and 3.0-3.5 mum in length. The temperature range for growth was 25-75 degrees C, with an optimum at 60 degrees C. The pH range for growth was 5.0-7.5, with an optimum at pH 6.5. Growth of strain SL50(T) was observed at NaCl concentrations ranging from 1.0 to 6.0 % (w/v) with an optimum at 2.5 % (w/v). The generation time under optimal growth conditions for strain SL50(T) was 60 min. Strain SL50(T) used molecular hydrogen, acetate, lactate, succinate, pyruvate and complex proteinaceous compounds as electron donors, and Fe(III), Mn(IV), nitrate or elemental sulfur as electron acceptors. The G+C content of the DNA of strain SL50(T) was 28.7 mol%. 16S rRNA gene sequence analysis revealed that the closest relative of strain SL50(T) was Deferribacter abyssi JR(T) (95.5 % similarity). On the basis of its physiological properties and phylogenetic analyses, the isolate is considered to represent a novel species, for which the name Deferribacter autotrophicus sp. nov. is proposed. The type strain is SL50(T) (=DSM 21529(T)=VKPM B-10097(T)). Deferribacter autotrophicus sp. nov. is the first described deep-sea bacterium capable of chemolithoautotrophic growth using molecular hydrogen as an electron donor and ferric iron as electron acceptor and CO(2) as the carbon source.
    International journal of systematic and evolutionary microbiology 07/2009; 59(Pt 6):1508-12. · 2.11 Impact Factor
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    ABSTRACT: Samples of water from the hot springs of Uzon Caldera with temperatures from 68 to 87 degrees C and pHs of 4.1 to 7.0, supplemented with proteinaceous (albumin, casein, or alpha- or beta-keratin) or carbohydrate (cellulose, carboxymethyl cellulose, chitin, or agarose) biological polymers, were filled with thermal water and incubated at the same sites, with the contents of the tubes freely accessible to the hydrothermal fluid. As a result, several enrichment cultures growing in situ on different polymeric substrates were obtained. Denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA gene fragments obtained after PCR with Bacteria-specific primers showed that the bacterial communities developing on carbohydrates included the genera Caldicellulosiruptor and Dictyoglomus and that those developing on proteins contained members of the Thermotogales order. DGGE analysis performed after PCR with Archaea- and Crenarchaeota-specific primers showed that archaea related to uncultured environmental clones, particularly those of the Crenarchaeota phylum, were present in both carbohydrate- and protein-degrading communities. Five isolates obtained from in situ enrichments or corresponding natural samples of water and sediments represented the bacterial genera Dictyoglomus and Caldanaerobacter as well as new archaea of the Crenarchaeota phylum. Thus, in situ enrichment and consequent isolation showed the diversity of thermophilic prokaryotes competing for biopolymers in microbial communities of terrestrial hot springs.
    Applied and Environmental Microbiology 11/2008; 75(1):286-91. · 3.95 Impact Factor
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    ABSTRACT: A moderately thermophilic, anaerobic bacterium (strain SG 508T) was isolated from a hydrothermal vent chimney located at 1 degrees N on the East Pacific Rise at a depth of 2650 m. Cells of strain SG 508T were straight to slightly curved rods, 0.4-0.6 microm in diameter and 2.0-3.0 microm in length. Spore formation was observed only below pH 5.5. The temperature range for growth was 22-60 degrees C, with optimum growth at 50 degrees C. The pH range for growth was 4.0-8.5, with optimum growth at pH 6.0-6.8. Growth of strain SG 508T was observed at NaCl concentrations ranging from 1.0 to 6.0 % (w/v), with optimum growth at 2.5 % (w/v). Substrates utilized by strain SG 508T included casein, peptone, tryptone, yeast extract, beef extract, starch, maltose and glucose. The products of glucose fermentation were ethanol, acetate, H2, formate and CO2. Strain SG 508T was able to reduce elemental sulfur to hydrogen sulfide. The DNA G+C content of strain SG 508T was 30.9 mol%. 16S rRNA gene sequence analysis revealed that the isolated organism belonged to cluster I of the genus Clostridium. On the basis of its physiological properties and data from phylogenetic analyses, strain SG 508T is considered to represent a novel species of the genus Clostridium, for which the name Clostridium tepidiprofundi sp. nov. is proposed. The type strain is SG 508T (=DSM 19306T =VKM B-2459T).
    International journal of systematic and evolutionary microbiology 05/2008; 58(Pt 4):852-5. · 2.11 Impact Factor
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    ABSTRACT: A moderately thermophilic, facultatively anaerobic bacterium capable of reducing Cr(VI) (strain SKC1) was isolated from municipal sewage. Based on the analysis of the 16S rRNA gene nucleotide sequence and DNA-DNA hybridization data, strain SKC1 was identified as a representative of the species Bacillus thermoamylovorans. B. thermoamylovorans SKC1 is capable of reducing chromate with L-arabinose as an electron donor with an optimum at 50 degrees C and neutral pH. The culture is able to reduce Cr(VI) at its initial concentration in the medium of up to 150 mg/l. In addition to chromate, strain SKC1 is capable of reducing selenite and tellurite, as well as soluble forms of Fe(III). It was shown that Cr(VI), Te(IV), and Se(IV) exert a bacteriostatic effect on strain SKC1, and the reduction of these anions performs the detoxification function. This is the first communication on the reduction of chromate, selenite, tellurite, and soluble Fe(III) species by a culture of thermophilic bacilli.
    Mikrobiologiia 01/2007; 76(5):602-7.
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    ABSTRACT: Overall, 30 strains of hyperthermophilic archaea, representing seven species of the genera Thermococcus, Desulfurococcus, Thermoproteus, and Acidilobus, were tested for the presence of thermostable DNA polymerases. Thermostabilities of the polymerases varied distinctly among the strains within one species. Polymerases of five strains retained 60–100% activity upon incubation of the preparations at 95C for 120 min. A new DNA polymerase was isolated from the strain Thermococcus litoralis Sh1AM, possessing the enzyme with the most promising properties, and characterized. Molecular weight of the enzyme is 90–100% kDa. The purified DNA polymerase preserved 50% of the initial activity upon incubation at 95C for 120 min. The polymerase isolated displayed an associated 3–5 exonuclease activity. The error rate when extending a DNA strand was at least twofold lower compared with Taq polymerase. The main physicochemical and enzymatic properties of the new polymerase are similar to the known DNA polymerases of family B.
    Applied Biochemistry and Microbiology 12/2004; 41(1):34-41. · 0.69 Impact Factor
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    ABSTRACT: A method for rapid detection and identification of hyperthermophilic archaea of the family Thermococcaceae based on PCR amplification of 16S rRNA gene fragments with primers TcPc 173F (5'-TCCCCCATAGGYCTGRGGTACTGGAAGGTC-3') and TcPc 589R (5'-GCCGTGRGATTTCGCCAGGGACTTACGGGC-3') was developed and used for identification of new isolates.
    Applied and Environmental Microbiology 10/2004; 70(9):5701-3. · 3.95 Impact Factor
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    ABSTRACT: A stable association of hyperthermophilic microorganisms (82C), which contained mostly cocci and a minor amount of non-spore-forming rods, was obtained from the digested sludge of an anaerobic digestor used to process municipal wastewater under thermophilic conditions (50C). PCR amplification of 16S rRNA genes using total DNA isolated from this association and archaea-specific primers, followed by sequencing of the product obtained, showed that the archaeal component was represented by a single nucleotide sequence, which was 99.9% homologous to the 16S rRNA gene of Sulfophobococcus zilligii. Thus, a hyperthermophilic archaeon was for the first time detected in a system of anaerobic biological treatment of wastewater. In addition, this is the first report on the detection of a culturable member of Crenarchaeota in anthropogenic habitats with neutral pH.
    Microbiology 01/2004; 73(5):616-620. · 0.65 Impact Factor
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    ABSTRACT: A stable association of hyperthermophilic microorganisms (82 degrees C), which contained mostly cocci and a minor amount of non-spore-forming rods, was obtained from the fermented sludge of a methane tank used to process municipal wastewaters under thermophilic conditions (50 degrees C). PCR amplification of 16S rRNA genes using total DNA isolated from this association and archaea-specific primers, followed by sequencing of the product obtained, showed that the archaeal component was represented by a single nucleotide sequence, which was 99.9% homologous to 16S rRNA gene of Sulfophobococcus zilligii. Thus, a hyperthermophilic archaeon was for the first time detected in a system of anaerobic biological treatment of wastewater. In addition, this is the first report on the detection of a cultivated member of Crenarchaeota in anthropogenic habitats with neutral pH.
    Mikrobiologiia 01/2004; 73(5):716-20.