The sequential combination of ultrasonication and ozonation as sewage sludge treatment prior to anaerobic digestion was investigated. Synergistic volatile suspended solids (VSS) solubilization was observed when low energy ultrasonication (⩽12kJg(-1) TS) was followed by ozonation. 0.048gO3g(-1) TS ozonation induced the maximum VSS solubilization of 41.3% when the sludge was pre-ultrasonicated at 9kJg(-1) TS; while, the same ozone dosage applied without prior ultrasonication only induced 21.1% VSS solubilization. High molecular weight (MW) components (MW>500kDa) were found to be the main solubilization products when sludge was only ozonated. However, solubilization products by ozone were mainly in the form of low MW components (MW<27kDa) when sludge was pre-ultrasonicated. The high MW products generated by ultrasound were effectively degraded in the subsequent ozonation. Anaerobic biodegradability increased by 34.7% when ultrasonication (9kJg(-1) TS) and ozonation (0.036gO3g(-1) TS) were combined sequentially. The maximum methane production rate increased from 3.53 to 4.32, 4.21 and 4.54mL CH4d(-1) after ultrasonication, ozonation and ultrasonication-ozonation pre-treatments, respectively.