The use of consecutive feeding was applied to investigate the response of the microbial biomass to a second addition of substrates in terms of biodegradation using batch tests as a promising alternative to predict the behavior of the process. Anaerobic digestion (AD) of the slaughterhouse waste (SB) and its co-digestion with manure (M), various crops (VC), and municipal solid waste were evaluated. The results were then correlated to previous findings obtained by the authors for similar mixtures in batch and semi-continuous operation modes. AD of the SB failed showing total inhibition after a second feeding. Co-digestion of the SB + M showed a significant improvement for all of the response variables investigated after the second feeding, while co-digestion of the SB + VC resulted in a decline in all of these response variables. Similar patterns were previously detected, during both the batch and the semi-continuous modes.