This study investigated the effects of ozone and ultrasound (US) pretreatments, both individually and combined, on waste activated sludge reduction. Batch tests were conducted first to optimize the individual ozone and US pretreatments. Maximum sludge reduction ratios of 10.89% and 23% were obtained at 0.15 g O-3/g total solids ozone dose and 1.5 W/mL US energy density, respectively. The combined ozone and US pretreatments were studied using response surface methodology. A maximum sludge reduction ratio of 40.14% was achieved by the combined ozone/US pretreatment with an ozone dose of 0.154 g O-3/g total solids and an US energy density of 1.445 W/mL. The analysis of the dissolved organic matter by three-dimensional excitation-emission matrix fluorescence spectroscopy showed that the combined pretreatment was superior to the individual ozone and US pretreatments, and also demonstrated the synergetic effect of these two combined pretreatments. (c) 2013 Elsevier Ltd. All rights reserved.