Coupling the activated sludge and the ozonation processes is an efficient, although expensive, solution for sludge reduction. A better knowledge of the mechanisms involved in the degradation of various sludge fractions by ozone is needed to optimize the coupled process. The objectives of this study were to determine the biodegradability of ozone-solubilized endogenous residue, the action of ozone on the active biomass and the solubilization yield of these two main sludge fractions. Batch tests were conducted with slug input of ozone stock solution into fresh or aerobically digested synthetic sludge. Biodegradability of the solubilized endogenous residue was increased by ozonation by up to 0.27 g BOD5/g CODi. Ozone caused biomass lysis, as opposed to an increase in maintenance needs, as shown by a correlation between the decrease in microbial activity and viability. Lysis caused by ozonation was associated with a solubilization of 20% of the lyzed cell COD mass. Solubilization yields were of 9.6 and of 1.9 to 3.6 g COD/g O3 for fresh and aerobically digested sludge, respectively. Design of sludge ozonation processes should account for the variability between the solubilization yield and biodegradability of the various sludge fractions.