A pilot-scale multi-staged UASB (MS-UASB) reactor with a working volume of 2.5 m3 was operated for thermophilic (55 degrees C) treatment of an alcohol distillery wastewater for a period of over 600 days. The reactor steadily achieved a super-high rate COD removal, that is, 60 kgCOD m(-3) d(-1) with over 80% COD removal. However, when higher organic loading rates were further imposed upon the reactor, that is, above 90 kgCOD m(-3) d(-1) for barely-based alcohol distillery wastewater (ADWW) and above 100 kgCOD m(-3) d(-1) for sweet potato-based ADWW, the reactor performance somewhat deteriorated to 60 and 70% COD removal, respectively. Methanogenic activity (MA) of the retained sludge in the thermophilic MS-UASB reactor was assessed along the time course of continuous run by serum-vial test using different substrates as a vial sole substrate. With the elapsed time of continuous run, hydrogen-utilizing MA, acetate-utilizing MA and propionate-fed MA increased at maximum of 13.2, 1.95 and 0.263 kgCOD kgVSS(-1) d(-1), respectively, indicating that propionate-fed MA attained only 1/50 of hydrogen-utilizing MA and 1/7 of acetate-utilizing MA. Since the ADWW applied herewith is a typical seasonal campaign wastewater, the influence of shut-down upon the decline in sludge MA was also investigated. Hydrogen-utilizing MA and acetate-utilizing MA decreased slightly by 3/4, during a month of non-feeding period, whereas propionate-fed MA were decreased significantly by 1/5. Relatively low values of propionate-fed MA and its vulnerability to adverse conditions suggests that the propionate degradation step is the most critical bottleneck of overall anaerobic degradation of organic matters under thermophilic condition.