The adaptation of an anaerobic culture (anaerobic sludge) to a specific substrate brings significant changes to its microbial population. These changes can be described by the sludge's ability to treat various substrates such as carbohydrates or proteins or "intermediate" products of anaerobic metabolism such as L-lactic, propionic, and acetic acids. The activity of the sludge with respect to a specific substrate is a critical parameter, because the anaerobic degradability of wastewaters depends strongly on it. This work examines and quantifies the differentiation of two anaerobic sludges of the same origin, following an adaptation period of about 18 months to lactose and gelatin, respectively. The acclimation has a significant effect on the maximum specific utilization rates of various compounds and on their apparent consumption kinetics. It is noticeable, however, that even if the anaerobic cultures were not exposed to a specific substrate for a prolonged period of time (more than a year), they still kept the ability of hydrolyzing or degrading it. In addition, the acclimation has an unquestionable effect on the stoichiometry of the production of volatile fatty acids and L-lactate. Finally, from codigestion experiments it is shown that codigestion of lactose and gelatin appears to have no effect on their hydrolysis kinetics in any of the lactose or gelatin acclimated cultures; specifically, the hydrolysis kinetics remained the same as calculated when lactose or gelatin were the only fed substrates. Similarly, the kinetics of L-lactate and D-glucose biodegradation seemed to be unchanged. On the other hand, codigestion has a significant effect on the production of L-lactic, propionic, and acetic acids, which can be attributed to the increased hydrogen production accompanying gelatin biodegradation.