Abstarct: Nosocomial infections are one of the most important causes of mortality worldwide, and the World Health Organization estimates that 15% of all hospitalized patients suffer from them. Adhesion of pathogen elements to surface and biofilm formation is an important biological issue for health services. Because of the variety of biofilms and their specific phenotype, the demolition of them requires rigorous treatments, which are mostly possible with chemical and oxidizing biocides. Concentrations of disinfectants to eliminate bacterial biofilms maybe 1000 times more than is required to eliminate planktonic bacteria of the same species, therefore, antimicrobial treatments are often not able to eliminate the biofilm alone. Thus control and elimination of biofilms can be achieved through the combined use of treats with different spectra and conditions. The use of new and uncomplicated interfering agents to enhance the synergistic effect of disinfectants plays an important role in controlling the risk of biofilm compounds in the environment. The present study aimed to investigate the biofilm and synergistic effects of three disinfectants of SaniHigene, SAYASEPT-HP, and Bellasan in prevention and eliminating the bacterial species Escherichia coli, Enterococcus faecalis, Pseudomonas aeruginosa, and Staphylococcus aureus by Micro-nano bubble (MNB) technology. The antibacterial effect of disinfectants and Ozone Micro Nano Bubbles (OMNB) with a concentration of 5 g/h was determined by MIC and MBC methods. To contrast, disinfectants were diluted with distilled water and ozone-containing water with a maximum concentration of 2% by volume. The results showed that the synergistic effect of OMNB and SaniHigene decreased 1 dilution in MIC of Pseudomonas aeruginosa compared to dilution with distilled water. The synergistic effect of OMNB and SAYASEPT-HP caused 1 dilution in MIC of Escherichia coli and Enterococcus faecalis and 1 dilution in MBC of Escherichia coli, Enterococcus faecalis, and Staphylococcus aureus. The synergistic effect of OMNB and Bellasan decreased 1 dilution in MIC of Pseudomonas aeruginosa and Staphylococcus aureus and 1 dilution in MBC of Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa. Microtiter Plate Mueller was used to determine the synergistic effect of OMNB on the potency of disinfectants used in inhibiting bacterial biofilm. Statistical analyzes on Optical Density (OD) showed that the biofilms of Escherichia coli, Enterococcus faecalis, Staphylococcus aureus, and Pseudomonas aeruginosa due to the interfering effect of ozone were 68.86%, 43.15%, 55.6%, and 62.12% in SaniHigene, 75.45%, 41.42%, 67.01% and 39.68% in SAYASEPT-HP and 70%, 48.87%, 63.07% and 55.47% in Bellasan Was restrained. Also, the synergistic effect of OMNB and SaniHigene caused a plus decrease in the biofilm strength of Staphylococcus aureus and Pseudomonas aeruginosa from strong to moderate compared to dilution with distilled water. The synergistic effect of OMNB and Bellasan decreased a plus in the biofilm strength of Enterococcus faecalis and Pseudomonas aeruginosa from strong to moderate. According to the results of this study, the use of OMNB in diluting disinfectants has a higher anti-biofilm effect and also decreases the concentration of disinfectants to control and eliminate these bacteria.