Anaerobic digestion leads to significant changes of the sludge structural matrix, affecting particle size distribution and dewaterability. The surface charge, determined by means of streaming current, can be effectively used to monitor the complex phenomena of floc disruption, colloid formation and chemical conditioning. To study the relation between surface charge and optimal dosage, two different cationic polyelectrolytes were used: Praestol 644, polymer with high molecular weight and low charge density, and Poly Dadmac, with relatively low molecular weight but high charge density. The optimal Poly Dadmac dosage strictly met the value required to neutralise particle charge whereas the optimal dosage of Praestol 644 indicated that the relevant charge was considerably lower than the one required for charge neutralisation. Mechanisms of action are therefore clearly different. Another objective was to investigate the changes of dewatering characteristics of secondary sludge during anaerobic digestion tests at different inoculum content by determining charge density, and optimal polymer dosage. The optimal polyelectrolyte dosage remains almost constant during digestion at high inoculum, but a significant increase in the first period is observed at low inoculum, thus suggesting that the release of colloidal and supracolloidal material from sludge affects dewaterability, especially in the first days of digestion.