| Summary: | 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<sup>−1</sup>, 100 Hz, 5000 pulses in trains mode of 500 pulses with a pulse duration of 10 µs) on <i>Saccharomyces cerevisiae</i> 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<i> S. cerevisiae</i> were suspended in PBS, increasing the current density from 0.02 to 3.3 A cm<sup>−2</sup> (corresponding to a total specific energy of 22.04 to 614.59 kJ kg<sup>−1</sup>) led to an increase of <i>S. cerevisiae</i> eradication. At 3.3 A cm<sup>−2</sup>, a total <i>S. cerevisiae</i> eradication was observed. However, when the <i>S. cerevisiae </i>in PBS was treated with the highest current density of 3.3 A cm<sup>−2</sup>, 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 <i>S. cerevisiae</i> 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 <i>S. cerevisiae</i> were suspended in YMB containing tartaric acid (pH 3.4) and ethanol to a final concentration of 10% (<i>v/v</i>), 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 SO<sub>2</sub>, 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.
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