Selected operational conditions of activated sludge are investigated in relation to their effect on flocculation and dewatering. Semi-continuous reactors with 2 -L volume were operated at different mean cell residence times, C/N ratios, and three different influent cations. Results show that MCRT, C/N ratio, and the cation type affect the bioflocculation capacity of activated sludge measured by the quantity of extracellular polymeric substances. As the MCRT value operated in the reactors and the C/N ratio of the influent wastewater increase, total amount of polymers produced increases. High MCRT values and low C/N values cause good dewatering of the sludge. All cations are shown to stimulate the EPS production in a way that the highest total EPS concentrations are observed at the highest cation dosages. The dewaterability of the sludges improves only with increasing calcium and magnesium concentrations and deteriorates with increasing potassium concentration.