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The cladodes of Opuntia ficus-indica are widely consumed across the world and appreciated for their beneficial effects. The present work aims to study their phytochemical profile and evaluate the antibacterial potential of the methanolic extract of O. ficus-indica. The antibacterial effect was determined using agar disk diffusion and minimum inhibitory concentration methods. The quantitative evaluation showed that O. ficus-indica were rich in polyphenols, with predominant levels of flavonoids (11.86 ± 0.10 mg QE/g) and condensed tannins (6.45 ± 0.22 mg CE/g). The results clearly demonstrated that O. ficus-indica extract had interesting antibacterial potential against positive and negative strains, with inhibition diameters ranging from 9 to 18 mm. As a second parameter, minimum inhibitory concentration was recorded as 100 mg/ml for S. aureus and P. aeruginosa. O. ficus-indica extracts may serve as a potential source of antimicrobials against infectious agents.