BACKGROUND AND OBJECTIVES: The most common cause of healthcare-associated diarrhea is infection (CDI), which causes severe and recurring symptoms. The increase of antibiotic-resistant requires alternate treatments. Postbiotics, metabolites produced by probiotics, fight CDI owing to their antibacterial capabilities. This study aims to evaluate the antibacterial, antibiofilm, and anti-toxigenic potential of postbiotics in combating CDI. MATERIALS AND METHODS: GC-MS evaluated postbiotics from and . Disk diffusion and broth microdilution determined antibacterial inhibition zones and MICs. Microtiter plates assessed antibiofilm activity. MTT assay evaluated postbiotics anti-viability on HEK293. ELISA testing postbiotic detoxification of toxins A and B. Postbiotics were examined for and genes expression using real-time PCR. RESULTS: The most identified and postbiotic compounds were glycolic acid (7.2%) and butyric acid (13.57%). and displayed 13 and 10 mm inhibition zones and 2.5 and 5 mg/ml MICs against reduced biofilm at 1.25 mg/ml by 49% and by 31%. MTT assay showed both postbiotics had little influence on cell viability, which was over 80%. The detoxification power of postbiotics revealed that decreased toxin A and B production more effectively than , and also their related and genes expression reduction were statistically significant (p < 0.05). CONCLUSION: Postbiotics' ability to inhibit bacterial growth, biofilm disruption, and toxin reduction makes them a promising adjunctive for CDI treatment and a good solution to pathogens' antibiotic resistance.