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The use of plants in the biological production of silver nanoparticles for antibacterial applications is a growing field of research. In the current work, we formulated Ocimum kilimandscharicum extracts using silver nanoparticles, and evaluated its potential antibacterial activity. Aqueous and methanol plant extracts were used to reduce silver nitrate at different time intervals (30 to 150 minutes) and pH (2 to 11). The UV-visible absorption spectrum recorded for methanol and aqueous extracts revealed a successful synthesis of AgNPs for methanol and aqueous extracts. The antimicrobial activity of the AgNPs was evaluated against Escherichia coli ATCC 25922, Salmonella choleraesuius ATCC 10708, and Staphylococcus aureus ATCC 25923 The best inhibition zone for the methanol and aqueous-mediated AgNPs, ranging from 12 ± 1 to 16 ± 1mm. Additionally, the methanol and aqueous extract silver nanoparticles had the same Minimum Inhibitory Concentration (6.25 ± 0.00 mg/ml), whereas the Minimum Bactericidal Concentrations were 12.5 ± 0.00 and 25 ± 0.00 mg/ml, respectively. The highest inhibition zone of 16 ± 1 mm was observed against Salmonella choleraesuius with 50 ± 0.00 mg/ml aqueous silver nanoparticles. The results show that the silver nanoparticles made with Ocimum kilimandscharicum have antibacterial action against those microorganisms.