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As many as 99% of illnesses caused by Listeria monocytogenes are foodborne in nature, leading to 94% hospitalizations, and are responsible for the collective annual deaths of 266 American adults. The current study is a summary of microbiological hurdle validation studies to investigate synergism of mild heat (up to 55 °C) and elevated hydrostatic pressure (up to 380 MPa) for decontamination of Listeria monocytogenes and natural background microflora in raw milk and phosphate-buffered saline. At 380 MPa, for treatments of 0 to 12 min, d-values of 3.47, 3.15, and 2.94 were observed for inactivation of the pathogen at 4, 25, and 50 °C. Up to 3.73 and >4.26 log CFU/mL reductions (p < 0.05) of habituated Listeria monocytogenes were achieved using pressure at 380 MPa for 3 and 12 min, respectively. Similarly, background microflora counts were reduced (p < 0.05) by 1.3 and >2.4 log CFU/mL after treatments at 380 MPa for 3 and 12 min, respectively. Treatments below three min were less efficacious (p ≥ 0.05) against the pathogen and background microflora, in the vast majority of time and pressure combinations. Results of this study could be incorporated as part of a risk-based food safety management system and risk assessment analyses for mitigating the public health burden of listeriosis.