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

Abstract

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.