Quantification of the inhibitory effect of methyl fluoride on methanogenesis in mesophilic anaerobic granular systems

The inhibitory effect of CH₃F on methanogenesis in mesophilic anaerobic granules was tested at different concentrations (0-10% v/v, in the gas phase) and verified by the stable carbon isotopic signatures of CH₄ and CO₂. The results showed that the inhibitory effect increased with the initial CH₃F concentration up to 5%. The CH₃F concentration causing 50% metabolic inhibition was 0.32%. Complete inhibition of acetoclastic methanogenesis with a 91% reduction in total methanogenic activity was achieved when 5% CH₃F was initially added to the headspace, which resulted in 870 μM dissolved CH₃F in the liquid. It was much higher than that applied in other natural anoxic non-granular systems, indicating that the layered granular structure influenced the inhibitory effect. The obvious increase in hydrogen content indicated that high concentrations of CH₃F (?5%) suppressed hydrogenotrophic methanogenesis as well. The stable inhibition lasted for at least 6 d as the CH₃F concentration decreased slowly with incubation time. These results suggested that CH₃F could be used for investigating methanogenic processes in anaerobic granular systems after the CH₃F concentration and incubation time for specific inhibition of acetoclastic methanogenesis were carefully determined. In the present system, CH₃F concentration of 5% was suggested to be optimal.