The aim of this study was to compare the biocidal effectiveness of disinfectants solutions prepared with ozonated and non-ozonated water against Listeria monocytogenes. Six L. monocytogenes strains were the research material (four isolates from food: meat (LMO-M), dairy products (LMO-N), vegetables (LMO-W), and fish (LMO-R); one clinical strain (LMO-K) and reference strain ATCC 19111). The evaluation of the biocidal effectiveness of disinfectant solutions (QAC—quaternary ammonium compounds; OA—oxidizing agents; ChC—chlorine compounds; IC—iodine compounds; NANO—nanoparticles) was carried out, marking the MBC values. Based on the obtained results, the effectiveness coefficient (A) were calculated. The smaller the A value, the greater the efficiency of disinfection solutions prepared on the basis of ozonated versus non-ozonated water. Ozonated water showed biocidal efficacy against L. monocytogenes. Among tested disinfectentants, independent on type of water used for preparation, the most effective against L. monocytogenes were: QAC 1 (benzyl-C12-18-alkydimethyl ammonium chlorides) (1.00 × 10−5–1.00 × 10−4 g/mL) in quaternary ammonium compounds, OA 3 (peracetic acid, hydrogen peroxide, bis (sulphate) bis (peroxymonosulfate)) (3.08 × 10−4 –3.70 × 10−3 g/mL) in oxidizing agents, ChC 1 (chlorine dioxide) (5.00 × 10−8 –7.00 × 10−7 g/mL) in chlorine compounds, IC 1 (iodine) (1.05–2.15 g/mL) in iodine compounds, and NANO 1 (nanocopper) (1.08 × 10−4 – 1.47 × 10−4 g/mL) in nanoparticles. The values of the activity coefficient for QAC ranged from 0.10 to 0.40, for OA—0.15–0.84, for ChC—0.25–0.83, for IC—0.45–0.60, and for NANO—0.70–0.84. The preparation of disinfectants solution on the basis of ozonated water, improved the microbicidal efficiency of the tested disinfectant, especially the quaternary ammonium compounds. An innovative element of our work is the use of ozonated water for the preparation of working solutions of the disinfection agents. Use ozonated water can help to reduce the use of disinfectant concentrations and limit the increasing of microbial resistance to disinfectants. This paper provides many new information to optimize hygiene plans in food processing plants and limit the spread of microorganisms such as L. monocytogenes.