A key parameter in water and wastewater treatment technology is the biomass activity in terms of substrate removal ability. The effects of organic load rate and percentage of bed expansion on biofilm specific methanogenic activity were determined in an anaerobic fluidized bed reactor treating wine-distillery wastes in the thermophilic range (55°C). The proposed activity tests are highly reproducible: an experiment with three identical tests has shown that the standard deviation with respect to the mean values is less than 3%. Specific tests are applied to measure the maximum methanogenic activities of the biomass carrier in lab-scale anaerobic biofilm reactors. These tests have been successfully applied for monitoring the support colonization process and the evolution of biofilm activity in reactors, anaerobic filter and fluidized bed, with different operating conditions. The results show a dependence between the percentage of bed expansion and the specific activity of methanogenic microbiote on biofilm. There is a relationship between the percentage of bed expansion, the shear stress on the biofilm and the hydrodynamic conditions in the system. Initial biofilm detachment can be compensated with the increase of biomass and of its activity due to the reduction of the substrate diffusional limitations to the microorganism growth inside the support pores.