No abstract

One of the promising technologies that can inactivate microorganisms without heat is pulsed electric field (PEF) treatment. The aim of this study was to examine the influence of PEF treatment (2.9 kV cm−1, 100 Hz, 5000 pulses in trains mode of 500 pulses with a pulse duration of 10 µs) on Saccharomyces cerevisiae eradication and resealing in different conditions, such as current density (which is influenced by the medium conductivity), the sort of medium (phosphate buffered saline (PBS) vs. yeast malt broth (YMB) and a combined treatment of PEF with the addition of preservatives. When the S. cerevisiae were suspended in PBS, increasing the current density from 0.02 to 3.3 A cm−2 (corresponding to a total specific energy of 22.04 to 614.59 kJ kg−1) led to an increase of S. cerevisiae eradication. At 3.3 A cm−2, a total S. cerevisiae eradication was observed. However, when the S. cerevisiae in PBS was treated with the highest current density of 3.3 A cm−2, followed by dilution in a rich YMB medium, a phenomenon of cell membrane resealing was observed by flow cytometry (FCM) and CFU analysis. The viability of S. cerevisiae was also examined when the culture was exposed to repeating PEF treatments (up to four cycles) with and without the addition of preservatives. This experiment was performed when the S. cerevisiae were suspended in YMB containing tartaric acid (pH 3.4) and ethanol to a final concentration of 10% (v/v), which mimics wine. It was shown that one PEF treatment cycle led to a reduction of 1.35 log10, compared to 2.24 log10 when four cycles were applied. However, no synergic effect was observed when the preservatives, free SO2, and sorbic acid were added. This study shows the important and necessary knowledge about yeast eradication and membrane recovery processes after PEF treatment, in particular for application in the liquid food industry.

Pulsed electric fields (PEFs) technology was reported to be useful as a disinfection method in the liquid food industry. This technology may lead to membrane permeabilization and bacterial death. However, resuscitation of viable but non-culturable cells and sublethally injured microorganisms in food was reported to be associated with foodborne outbreaks. The main aim of this study was to investigate the possible recovery of injured PEF-treated bacteria. The PEF treatment of Staphylococcus aureus and Pseudomonas putida led to a reduction of 3.2 log10 and 4.8 log10, respectively. After 5 h, no colony forming units (CFUs) were observed when the bacteria were suspended in phosphate buffer saline (PBS); and for 24 h, no recovery was observed. The PEF-treated S. aureus in brain-heart infusion (BHI) medium were maintained at 1.84 × 104 CFU mL−1 for about 1.5 h. While P. putida decreased to zero CFU mL−1 by the 4th hour. However, after that, both bacteria recovered and began to multiply. Flow cytometry analysis showed that PEF treatment led to significant membrane permeabilization. Mass spectrometry analysis of PEF-treated P. putida which were suspended in BHI revealed over-expression of 22 proteins, where 55% were related to stress conditions. Understanding the recovery conditions of PEF-treated bacteria is particularly important in food industry pasteurization. To our knowledge, this is the first comprehensive study describing the recovery of injured PEF-treated S. aureus and P. putida bacteria.