Different methods were tested to evaluate the performance of a pretreatment before anaerobic digestion. Besides conventional biochemical parameters, such as the biochemical methane potential (BMP), the methane production rate, or the extent of solubilization of organic compounds, methods for physical characterization were also developed in the present work. Criteria, such as the particle size distribution, the water retention capacity, and the rheological properties, were thus measured. These methods were tested on samples taken in two full-scale digesters operating with cattle manure as a substrate and using hammer mills. The comparison of samples taken before and after the pretreatment unit showed no significant improvement in the methane potential. However, the methane production rate increased by 15% and 26% for the two hammer mills, respectively. A relevant improvement of the rheological properties was also observed. This feature is likely correlated with the average reduction in particle size during the pretreatment operation, but these results needs confirmation in a wider range of systems.