Synergistic Processing of Skim Milk with High Pressure Nitrous Oxide,Heat, Nisin,and Lysozyme to Inactivate Vegetative and Spore-Forming Bacteria

Individual and combined effects of high pressure nitrous oxide (HPN₂O), heat, and antimicrobials on the inactivation of Escherichia coli, Listeria innocua, and Bacillus atrophaeus endospores in milk were all evaluated after 20-min treatments. Stand-alone milk treatments with HPN₂O (15.2 MPa), heat (45 and 65 °C), or nisin (50 and 150 IU mL^?1) resulted in log₁₀ reductions ranging only from 0.1 to 2.1 for E. coli and L. innocua. Combining HPN₂O (15.2 MPa) with heat (65 °C) inactivated 6.0 and 5.1 log₁₀ in the vegetative bacteria, respectively. Similarly, reductions of 5.9 and ≥ 6.0 log₁₀ of respective E. coli and L. innocua cells in milk were achieved through a combination of HPN₂O (15.2 MPa), heat (65 °C), and nisin (150 IU mL^?1). A 2.5 log₁₀ cycle inactivation of spores was obtained by HPN₂O, nisin (at both 50 and 150 IU mL^?1), and lysozyme (50 μg mL^?1) at 85 °C. Combining these processing techniques resulted in significantly greater microbial inactivation (p < 0.05) than the sum of individual reductions from each treatment alone, indicating synergistic effects. HPN₂O irrespective of processing temperatures did not cause any occurrence of sub-lethally injured cells or disruption in colloidal stability of milk at 65 and 85 °C (p ≥ 0.05). Color and pH changes in milk following the most demanding treatment conditions were minimal.