To use the selective inhibition method for quantitative analysis of acetate metabolism in methanogenic systems,the responses of microbial communities and metabolic activities,which were involved in anaerobic degradation of acetate,to the addition of methyl fluoride(CH3F),2-bromoethanesulfonate(BES)and hydrogen were investigated in a thermophilic batch experiment.Both the methanogenic inhibitors,i.e.,CH3F and BES,showed their effectiveness on inhibiting CH4 production,whereas acetate metabolism other than acetoclastic methanogenesis was stimulated by BES,as reflected by the fluctuated acetate concentration.Syntrophic acetate oxidation was thermodynamically blocked by hydrogen(H2),while H2-utilizing reactions as hydrogenotrophic methanogenesis and homoacetogenesis were correspondingly promoted.Results of PCR-DGGE fingerprinting showed that,CH3F did not influence the microbial populations significantly.However,the BES and hydrogen notably altered the bacterial community structures and increased the diversity.BES gradually changed the methanogenic community structure by affecting the existence of different populations to different levels,whilst H2 greatly changed the abundance of different methanogenic populations,and induced growth of new species.
The impacts of landfill leachate irrigation on methane oxidation activities and methane-consuming bacteria populations were studied by incubation of landfill cover soils with leachate and (NH4)2SO4 solutiort at different ammonium concentrations. The community structures and abundances of methane-oxidizing bacteria (MOB) and ammonia-oxidizing bacteria (AOB) were examined by PCR- DGGE and real-time PCR. Compared with the pure (NH4)2SO4 solution, leachate addition was found to have a positive effect on methane oxidation activity. In terms of the irrigation amount, ammonium in leachate was responsible for the actual inhibition of leachate. The extent of inhibitory effect mainly depended on its ammonium concentration. The suppression of the predominant methane- consuming bacteria, type I MOB, was responsible for the decreased methane oxidation activity by ammonium inhibition. Methane- consuming bacteria responded diversely in abundance to ammonium. The abundance of type I MOB decreased by fivefold; type II MOB showed stimulation response of fivefold magnification upon the first addition but lessened to be lower than the original level after the second addition; the amount of AOB was stimulated to increase for 20-30 times gradually. Accumulated nitrate from nitrification strengthened the ammonium inhibition on type I and type II MOB, as a result, repetitive irrigation was unfavorable for methane oxidation.