Non-thermal processing of milk can potentially reduce nutrient loss, and a low-temperature-high-pressure (LTHP) treatment is considered as a promising alternative to thermal treatment, attracting considerable attention in recent years. The effect of LTHP treatment (−25 °C, 100–400 MPa) on the phase transition behavior of frozen milk was evaluated. The lethal and injured effects of different pressures and cycle numbers on E. coli in frozen milk were studied by using selective and non-selective enumeration media. Results from the gathered transient time–temperature–pressure data showed that pressures over 300 MPa could induce a phase transition from Ice I to Ice III. The treatment at −25 °C and 300 MPa could achieve a lethal effect similar to the two-cycle treatment of 400 MPa at room temperature. This meant that LTHP conditions can lower the operating pressure by at least 100 MPa or reduce the operation from two cycle to one cycle. Increasing the number of pressure cycles enhanced the lethal effects, which was not additive, but resulted in a transformation of part of the injured cells into dead cells. Transmission electron microscopy (TEM) and scanning electron microscopy (SEM) provided direct evidence for the breakdown of cell membrane and cell walls by phase transitions. Combined with a designed internal cooling device, the LTHP process can be expected to be a more attractive alternative to non-thermal processing for the dairy industry.

It was demonstrated that the inactivation of high pressure (HP) treatment on Escherichia coli survival in sterile physiological saline (SPS) was influenced by the treatment conditions: unfrozen, frozen-thawed and fully frozen (phase transition). In order to probe the enhanced phase transition microbial destruction, vibration effects of phase transition position were created and discussed. Test samples were placed in HP chamber for treatment (150/240/330 MPa, no holding time) at room temperature and a special cooling device was used to maintain the phase transition conditions. Results showed that the phase transition from ice I to ice III of frozen SPS could be realized based on the cooling of a 20% sodium chloride solution. HP treatment under fully frozen conditions produced the best lethal effect compared to unfrozen and freeze-thaw samples. Vibration tests were carried out by using model solutions and apple juice to explore the behavior of phase transition. A synchronous and advance phase transition of internal apple juice was realized, respectively, by using pure water and 5% sodium chloride solution as external vibration sources, and the advance phase transitions of external pure water were realized by using 5% sodium chloride solution and 5% glucose solution as internal vibration sources.