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To investigate the effect of ultrasonic pretreatment on anaerobic digestion and sludge dewaterability and further to probe into the influencing factors on sludge dewaterability,sludge flocs were stratified into four fractions:(1) slime; (2) loosely bound extracellular polymeric substances (LB-EPS); (3) tightly bound EPS (TB-EPS); and (4) EPS-free pellets.The results showed that ultrasonic pretreatment increased the anaerobic digestion efficiency by 7%-8%.Anaerobic digestion without ultrasonic pretreatment deteriorated the sludge dewaterability,with the capillary suction time (CST) increased from 1.42 to 47.3 (sec.L)/g-TSS.The application of ultrasonic pretreatment firstly deteriorated the sludge dewaterability (normalized CST increased to 44.4 (sec.L)/g-TSS),while subsequent anaerobic digestion offset this effect and ultimately decreased the normalized CST to 23.2 (sec.L)/g-TSS.The dewaterability of unsonicated sludge correlated with protein (p = 0.003) and polysaccharide (p = 0.004) concentrations in the slime fraction,while that of sonicated sludge correlated with protein concentrations in the slime and LB-EPS fractions (p < 0.05).Fluorescent excitationemission matrix analysis showed that the fluorescence matters in the LB-EPS fraction significantly correlated with sludge dewaterability during anarobic digestion.