Ultralow temperature high pressure processing enhances inactivation of norovirus surrogates.

High pressure processing (HPP) is a powerful non-thermal method for inactivating pathogens. Human norovirus and genetically-related caliciviruses are moderately sensitive to temperatures above 0 °C with >400 MPa (MPa) or higher required to inactivate multiple logs of virus. Sensitivity of murine norovirus (MNV) and Tulane virus (TV) to ice phase transitions was evaluated using ultra low temperature HPP. Identical samples containing MNV or TV were either equilibrated to +1.5 °C (thawed) or -40 °C (frozen) 24 h prior to pressurization. All samples (thawed and frozen) were then placed in a pre-chilled chamber which was then rapidly filled with -40 °C chamber fluid. Samples were immediately pressurized for 5 min at 200, 250 or 300 MPa. Controls were not pressurized. For samples that were thawed and then pressurized in 40 °C chamber fluid, the MNV average log reduction at 200 MPa was 4.4, while >6.1 log reduction (non-detectable) was achieved at 250 and 300 MPa. TV samples averaged 2.3, 5 and 4.3 log reduction at 200, 250, and 300 MPa respectively. For samples that were frozen and then pressurized in 40 °C chamber fluid, the MNV average log reductions were 2.3, 3.2 and 4.2 at 200 MPa, 250 MPa and 300 MPa, respectively, while TV samples averaged 0.81, 2.3 and 1.7 log reductions at 200, 250, and 300 MPa, respectively. Inactivation of TV within oysters at these pressures was also demonstrated. Overall, results indicate that in addition to enhancing inactivation of norovirus surrogates compared to higher temperatures, ultra-cold HPP performed on thawed samples especially enhances inactivation.