“…Methane was generated in all enrichments with the addition of ethanol after an initial lag phase of about 80 days, and 1.25, 327, 308, and 204 µmol of methane were accumulated in the treatments of S0, S30, S60, and S90, respectively, after 230 days of anaerobic incubation. The methane production rate and total amount of methane in the enrichments with the amendment of ethanol and bicarbonate (28.4 µmol CH 4 day −1 l −1 , 26.8 µmol CH 4 day −1 l −1 , and 17.7 µmol CH 4 day −1 l −1 in S30, S60, and S90, respectively) were much higher than those in S0 without bicarbonate, and they were much higher than previously reported biological methane production rates of 1.3-80 nmol CH 4 day −1 l −1 in subsurface petroleum reservoirs [27]. Maximum methane production occurred in S30, followed by S60, S90, and S0, which showed a similar trend with the amounts of consumed ethanol in the corresponding cultures, indicating that ethanol metabolism was directly or indirectly related to activities of methanogenesis under the experimental conditions.…”