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The pathogenic and spoilage microorganisms affect human health directly or indirectly and become increasingly important in most countries. Novel antimicrobial agents and disinfectants have become necessary due to the rise of antibiotic resistance phenomena. The current study investigate the inhibitory infect of ANOSAN on pathogenic bacteria and toxigenic fungi. Chemical analysis was performed for ANOSAN. The antimicrobial activity of ANOSAN was carried out using agar well diffusion assay and mean growth inhibition percentage and Minimum Inhibitory Concentration (MIC). The antibiofilm activity was conducted using Biofilm formation assay and the impact of the disinfectant on the preformed biofilm was visualized by Scanning Electron Microscope (SEM). The chemical composition of ANOSAN comprised very low amount of Sodium (0.034%), Chloride (0.01%), Hypochlorite (0.014%) and Organic matter (0.0062%). ANOSAN low concentration showed a broad spectrum of antibacterial activities against Gram positive, Gram negative bacteria and fungi. The MIC (0.781 mg/ml) was observed for Vibrio cholerae, Pseudomonas aeruginosa and Staphylococcus aureus. Sub-inhibitory concentrations of ANOSAN have successfully inhibited biofilm formation of the above mentioned bacterial pathogens. The ANOSAN appeared to be effective as bactericidal against Ps. aeruginosa and Staph. aureus. The time kill assay was observed between (0-6h) and between (0-3h) of exposure for both pathogens, respectively, and the viable bacterial counts remained undetectable after the previous time in a confirmatory experiment. Scanning Electron Microscopy (SEM) was conducted to confirm the antimicrobial activity of ANOSAN against Staph. aureus and Ps. aeruginosa. These data directly illustrate the destructive effects of the ANOSAN on the pathogenic bacteria. This work clarified that ANOSAN water had bactericidal and fungicidal activity along with inhibiting the ability of pathogenic bacteria to form biofilms, thus providing a good alternative to the use of traditional antimicrobials in food industry, pharmaceutical and medical applications. [ABSTRACT FROM AUTHOR]