The emergence of antibiotic-resistant pathogens has necessitated the exploration of plant-based antimicrobial agents as alternative therapies. This study evaluates the antibacterial activity of Detarium senegalense stem bark extract against clinically significant bacterial species: Staphylococcus aureus, Salmonella species., and Bacillus species. The stem bark was collected from Daura Local Government Area of Katsina state, Nigeria. Clinical isolates were collected from Ahmadu Bello University Medical Centre then taken to Department of Microbiology for reconfirmation using standard microbiological methods. The stem bark was extracted using ethanol, and the antibacterial efficacy was assessed through the agar well diffusion method at concentrations of 100mg/ml, 50mg/ml, 25mg/ml, and 12.5mg/ml. Inhibition zones were recorded to determine the effectiveness of the extract, and the Minimum Inhibitory Concentration (MIC) was established. The extract demonstrated significant antibacterial activity, with notable inhibition zones against S. aureus and Salmonella species at 50mg/ml and 25mg/ml, respectively, indicating its potential as a natural antibacterial agent. Phytochemical analysis revealed the presence of bioactive compounds such as flavonoids, tannins, and alkaloids, which likely contribute to the observed antimicrobial effects. This study supports the potential use of Detarium senegalense stem bark as a source of bioactive compounds against pathogenic bacteria, providing a foundation for future research into its therapeutic applications and possible development as an alternative treatment for bacterial infections.