Different processing sequences of ultrasound and heat were applied for the inactivation of Staphylococcus aureus and Escherichia coli in milk. The changes in whey protein content, particle size, zeta potential, pH, viscosity, and color values were studied to evaluate the effects on the quality of milk. Results indicated ultrasound treatment (600 W, 5 min) with postheating (63 °C 5 min) reduced S. aureus and E. coli by 1.58 and 2.02 log cfu/ml, respectively, but yielded 0.26 and 0.19 log cfu/ml sublethal S. aureus and E. coli. Milk preheated to 63 °C (5 min) followed by ultrasound (600 W, 5 min) showed 1.17 and 1.68 log reductions for S. aureus and E. coli, respectively, and the sublethal S. aureus and E. coli were reduced to 0.16 and 0.09 log cfu/ml. The increased cavitation effects make the simultaneous treatment (600 W, 63 °C, 5 min) the most effective approach in killing S. aureus and E. coli, causing 1.67 and 2.16 log reductions, respectively, and producing negligible sublethal bacteria. The simultaneous treatment was also able to improve some physical–chemical properties of milk; smaller particle size and whiter color were achieved. However, whey protein and stability indices of milk were slightly deteriorated by this approach.
In food processing fields, thermosonication can serve as an alternative processing technique to thermal pasteurization. Especially, the simultaneous treatment of ultrasound and heat showed best inactivation effect for viable and sublethal cells, while maintaining the overall milk quality in the meantime. The results of this research may be useful for the treatment of various bacterial contamination and give the guidance for milk processing.