Methanogenic Bacteria from the Bondyuzhskoe Oil Field: General Characterization and Analysis of Stable-Carbon Isotopic Fractionation

Abstract

Selective enrichment culture techniques were employed to obtain mixed cultures of methanogenic rods and sarcina from surface flooding waters and deep subsurface (-1650 m) oil-bearing sedimentary rocks and formation waters sampled from an old oil field in the U.S.S.R. previously reported to display active biological methanogenesis. The methanogens were selectively isolated as colonies on agar petri dishes that were incubated in a novel container. The general cellular and growth features of three Methanobacterium isolates were determined. These strains grew optimally at 37 to 45°C in anaerobic pressure tube cultures with a doubling time of 16 to 18 h on H2-CO2 and proliferated as autotrophs. Acetate addition significantly enhanced the final cell yield. Growth of these strains was completely inhibited by either 0.6 g of sodium sulfide per liter or 31.0 of sodium chloride per liter, but growth was not inhibited by either 0.3 g of sodium sulfide per liter or 1.0 g of sodium sulfate per liter. One novel isolate, Methanobacterium sp. strain ivanov, was grown on H2-CO2, and the stable-carbon isotopic fractionations that occurred during synthesis of methane, cell carbon, and lipids were determined. The results of this study were used to examine the anomalous relationship between the isotopic and chemical compositions of natural gas occurring in the deep subsurface environment of the oil field.

Distribution and activity of methanogens and homoacetogens in deep granitic aquifers at Äspö Hard Rock Laboratory, Sweden

“…The concentration of dissolved hydrogen in these experiments was approximately 50 WM and this value did not exceed the maximum value measured in the groundwater (50 WM hydrogen), but were higher than the values measured in several discrete samples. The rates of methane formation obtained are similar to or higher than those observed in other aquatic systems such as a meromictic lake [47], freshwater sediments [48,49] and £ooded oil ¢eld groundwater [50]. The time needed for formation of the methane observed in A ë spo ë groundwater was calculated as the ratio of the methane concentrations to the methane formation rates.…”

CO2 and CH4 isotope compositions and production pathways in a tropical peatland

“…The α app is a measure of the isotopic difference between CO 2 and CH 4 , regardless of whether the substrate for CH 4 production is CO 2 or acetate, but it tends to be higher when more of the CH 4 is produced via HM (1.06 to 1.090) and lower when AM is more important (1.040 to 1.06) Hornibrook et al, 2000;Whiticar, 1999;. Experiments on pure cultures of methanogenic archaea reveal fractionation factors for HM (α CO2 ) ranging from 1.037 to 1.079 [Belyaev et al, 1983;Botz et al, 1996]. In environmental field samples, α CO2 has been measured by incubating soil with CH 3 F to inhibit AM and by using mass balance.…”

Microbial Methane From Methylphosphonate Isotopically Records Source

“…Carbon isotope fractionation between methane and methylphosphonate (MPn; 13 ε CH4/MPn ) for (a) two marine (HI00D01 and HTCC7221) and four freshwater (LM-5, LM-Y, LM-1, and 6-1) bacterial strains grown on oxygen at 30°C or (b) Pseudomonas stutzeri strain HI00D01 at three temperatures on nitrate. (c) A compilation of C-isotopic fractionation factors ( 13 ε CH4/substrate ) from this study (MPn) compared to anaerobic axenic culture experiments from CO 2 reduction, methanol reduction, or acetate disproportionation (Belyaev et al, 1983;Botz et al, 1996;Bryant, 1979;Fuchs et al, 1979;Games et al, 1978;Gelwicks et al, 1989;Goevert & Conrad, 2009;Krzycki et al, 1987;Londry et al, 2008;Min & Zinder, 1989;Penger et al, 2012;Penning et al, 2006;Rosenfeld & Silverman, 1959;Valentine et al, 2004).…”