Influence of Pressurization Rate and Pressure Pulsing on the Inactivation of Bacillus amyloliquefaciens Spores during Pressure-Assisted Thermal Processing

Abstract

Pressure-assisted thermal processing (PATP) is an emerging sterilization technology in which a combination of pressure (500 to 700 MPa) and temperature (90 to 120°C) are used to inactivate bacterial spores. The objective of this study was to examine the role of pressurization rate and pressure pulsing in enhancing PATP lethality to the bacterial spore. Bacillus amyloliquefaciens TMW 2.479 spore suspensions were prepared in deionized water at three inoculum levels (1.1 × 109, 1.4 × 108, and 1.3 × 106 CFU/ml), treated at two pressurization rates (18.06 and 3.75 MPa/s), and held at 600 MPa and 105°C for 0, 0.5, 1, 2, 3, and 5 min. Experiments were carried out using custom-fabricated, high-pressure microbial kinetic testing equipment. Single and double pulses with equivalent pressure–holding times (1 to 3 min) were investigated by using the spore suspension containing 1.4 × 108 CFU/ml. Spore survivors were enumerated by pour plating, using Trypticase soy agar after incubation at 32°C for 2 days. During short pressure–holding times (≤2 min), PATP treatment with the slow pressurization rate provided enhanced spore reduction over that of the fast pressurization rate. However, these differences diminished with extended pressure–holding times. After a 5-min pressure–holding time, B. amyloliquefaciens population decreased about 6 log CFU/ml, regardless of pressurization rate and inoculum level. Double-pulse treatment enhanced PATP spore lethality by approximately 2.4 to 4 log CFU/ml, in comparison to single pulse for a given pressure–holding time. In conclusion, pressure pulsing considerably increases the efficacy of PATP treatment against bacterial spores. Contribution of pressurization rate to PATP spore lethality varies with duration of pressure holding.