The biochemical characterization of 13 extracellular polymeric substances (EPS) producing bacterial strains were carried out by BIOLOG. The bacterial strains were cultured in sterilized sludge for EPS production. Flocculation and dewatering capabilities of produced EPS (broth, crude slime and capsular) were examined using kaolin suspension combined with calcium (150 mg of Ca2+/L of kaolin suspension). BIOLOG revealed that there were 9 Bacillus, 2 Serratia and 2 Yersinia species. Most of these bacterial strains had the capability to utilize wide spectrum of carbon and nitrogen sources. EPS concentration of more than 1 g/L was produced by most of the bacterial strains. Concentration of EPS produced by different Bacillus strains was higher than that of Serratia and Yersinia. Broth EPS revealed flocculation activity more than 75% for Bacillus sp.7, Bacillus sp.4 and Bacillus sp.6, respectively. Flocculation activity higher than 75% was attained using very low concentrations of broth EPS (1.12-2.70 mg EPS/g SS). (C) 2012 Elsevier Ltd. All rights reserved.