In this comparative study, the bacterial inactivation efficiency of plasma-activated water (PAW) generated by two distinct plasma reactors, one utilizing a nitrogen plasma jet electrode and the other a hybrid argon plasma reactor, was explored. The present study involved the assessment of antimicrobial activity against suspensions of three Gram-positive and three Gram-negative bacterial strains in their planktonic cell state. Bacterial suspensions were introduced into PAW five days after generation. Subsequently, the viability of the bacteria was assessed at various time intervals, specifically at 0.5, 1, 3, 5, 10, and 24 h, in order to evaluate the effect of inactivation. Structural changes in bacteria after PAW treatment were assessed using a scanning electron microscope (SEM). The physicochemical properties of PAW, including pH, conductivity, and concentrations of H2O2, NO2−, and NO3− during aging were measured. The present study demonstrated the effective inactivation of the tested bacterial strains by PAW. Gram-positive bacteria displayed greater resistance compared to Gram-negative species, with the lowest reductions in bacterial counts observed for B. cereus, and the highest for Escherichia coli O157:H7. Morphological damage was evident across all bacterial species examined. Physicochemical measurements showed slow decay of the reactive species in the aging process. This study illustrated the potential utility of PAW as an alternative disinfectant.