In this study the inactivation of vegetative Bacillus subtilis ATCC 9372 cells in response to high intensity pulsed electric fields (PEF) is presented. The effect of medium pH in a range between 5.5 and 7 as well as the addition of 5% (v/v) of ethanol was studied. Furthermore, it was investigated how a simultaneous compression of the microbial suspension can impact its susceptibility to PEF. The logarithmic inactivation of B. subtilis was independent of medium pH and progressively increased with rising field strengths. The extent of this enhancement strongly depends on the specific energy of the pulses which are released into the suspension. Applying 100 pulses at 2.1 kJ/kg a 4 log-cycle inactivation was obtained. By the addition of 5% (v/v) ethanol a further drastic increase in lethality was observed but only when the pH was reduced to 5.5. At pH 7 the presence of ethanol produced a higher resistance against PEF. A synergistic effect of high pressure was only observed when the compression time prior to PEF was extended to 10 min.