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The biogenic synthesis of silver nanoparticles (AgNPs) using Aloe fleurentiniorum extract and antibacterial activity of the synthesized AgNPs were investigated. Examinations using ultraviolet–visible spectrophotometry, fourier transform infrared spectroscopy, energy-dispersive X-ray spectroscopy, and transmission electron microscopy confirmed the efficacy of AgNP biosynthesis using A. fleurentiniorum extract. Purely prepared AgNPs were spherical and moderately sized. The antibacterial activity of AgNPs was evaluated through the agar well diffusion assay using Staphylococcus aureus, methicillin-resistant S. aureus, Escherichia coli, and Salmonella typhimurium. The biosynthesized AgNPs consistently displayed strong antibacterial activity.