Sterilization, alkaline-thermal and acid-thermal treatments were applied to different sludge solids concentrations (17.0; 22.4; 29.8; 37.3; 44.8 g/L, respectively) and the pre-treated sludge was used as raw material for Serratia sp.1 to produce extracellular polymeric substances (EPS). After 72 h of fermentation, total EPS of 2.3 and 3.4 g/L were produced in sterilized and alkaline-thermal treated sludge as compared to that of 1.5 g/L in acid-thermal treated sludge. Lower EPS were produced at relatively higher solids concentrations (37.3; 44.8 g/L). Broth, crude forms of capsular and slime EPS were extracted from fermented broths and used as conditioning agents by combining with 150 mg of Ca(2+)/L of kaolin suspensions. Maximum flocculation activity of 79.1% and increased dewatering by 52.2% was achieved using broth and crude capsular EPS, respectively. The results demonstrated that EPS having high flocculating capability could be produced using wastewater sludge as sole raw material.