Abstract

Abstract We investigated the stable carbon and hydrogen isotopic signatures of methane, CO2 and water during microbial formation of methane from mineral oil-related compounds in order to determine the variability in the methane signatures. The isotopic discrimination for carbon and hydrogen between substrate and methane was calculated and resulted in e C DIC - CH 4 26–60‰, e C substrate - CH 4 16–33‰, e H H 2 O - CH 4 257–336‰ and e H substrate - CH 4 174–318‰, respectively. These carbon and hydrogen isotope signatures fell in to a relatively narrow range, suggesting a coupling of fermentation with acetoclastic and CO2 reducing methanogenesis. In order to characterize the microbial consortia involved in the methanogenic degradation of hexadecane, a methanogenic enrichment culture was incubated with 1-13C–hexadecane and its biomass was analyzed for the pattern and isotopic signature of carboxylic acids. The highest extent of labelling was detected in the n-C17 fatty acid with methyl groups at C-4, presumably indicative of Syntrophus sp. To determine if the isotope composition of methane can be used as an indicator for methanogenesis during growth with oil-related compounds in field studies, we analyzed the isotope composition of methane in a confined mineral oil contaminated aquifer. The variability in carbon and hydrogen isotope composition was almost identical to the values obtained from enrichment cultures, thereby providing a tool for screening for microbial methane formation during hydrocarbon exploration.

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