Geochemical and Microbiological Evidence for Microbial Methane Production in Deep Aquifers of the Cretaceous Accretionary Prism.

Accretionary prisms are thick layers of sedimentary material piled up at convergent plate boundaries. Large amounts of anaerobic groundwater and methane (CH(4)) are contained in the deep aquifers associated with accretionary prisms. In order to identify microbial activity and CH(4) production processes in the deep aquifers associated with the Cretaceous accretionary prism in Okinawa Island, Japan, we performed geochemical and microbiological studies using anaerobic groundwater and natural gas (mainly CH(4)) samples collected through four deep wells. Chemical and stable hydrogen and oxygen isotope analyses of groundwater samples indicated that the groundwater samples obtained from each site originated from ancient seawater and a mixture of rainwater and seawater, respectively. Additionally, the chemical and stable carbon isotopic signatures of groundwater and natural gas samples suggested that CH(4) in the natural gas samples was of a biogenic origin or a mixture of biogenic and thermogenic origins. Microscopic observations and a 16S rRNA gene analysis targeting microbial communities in groundwater samples revealed the predominance of dihydrogen (H(2))-producing fermentative bacteria and H(2)-utilizing methanogenic archaea. Moreover, anaerobic cultures using groundwater samples suggested a high potential for CH(4) production by a syntrophic consortium of H(2)-producing fermentative bacteria and H(2)-utilizing methanogenic archaea through the biodegradation of organic substrates. Collectively, our geochemical and microbiological data support the conclusion that the ongoing biodegradation of organic matter widely contributes to CH(4) production in the deep aquifers associated with the Cretaceous accretionary prism.