Ultrasound treatment is often applied to enhance the anaerobic digestion of sludge. Optimal conditions for organic matter solubilisation of primary, secondary and mixed sludge were assessed by implementing ultrasound disruption at different specific energies (from 3,500 to 21,000 kJ/kgTS). The variation in soluble chemical oxygen demand (sCOD) and ammonium nitrogen (NH4+-N) was monitored following the treatment, and after a subsequent fermentation (24 h, 37 °C). The effect of the treatment was clearly more pronounced in secondary sludge than in the other types of sludge. Relatively minimal values in solubility were found when applying ultrasound at different energies depending on the sludge (3,500–7,000 kJ/kgTS in primary sludge and 10,500–14,000 kJ/kgTS in secondary sludge). This minimal value was not so noticeable in mixed sludge. The addition of inoculum was not required after ultrasound disruption in order to perform the subsequent fermentation. After this final stage, no general pattern in terms of sCOD was observed. Increases and decreases were conditioned by the coverage of the ultrasound irradiation; NH4+-N values increased notably during the fermentation.