This plant has been considered a medicinal plant in Yemen's traditional medicine, used as an antiseptic for wounds. This present study was carried out to evaluate the antibacterial properties of hexane, chloroform, butanol, and aqueous extracts of Jatropha spinosa latex. Agar disc diffusion tests were carried out to determine the antimicrobial effects of the Jatropha spinosa latex extracts against Gram-positive (B. cereus, S. aureus, and S. pyogenes), Gram-negative (Klebsiella, E. coli, and P. aeruginosa) bacterial strains. Among the various extracts, the hexane extract of Jatropha spinosa latex has the strongest antibacterial potency against Staphylococcus aureus (gram-positive), and the Butanol extract showed the highest effect on S. aureus, S. pyogenes, and Klebsiella at a higher concentration of 1mg/1ml with zones of inhibition 11.26mm, 11.53mm, and 11.6mm, respectively. Water extract showed the maximum (11.13± 1.02mm) inhibitory effect against Klebsiella at a higher concentration of 1mg/1ml, and no activity was observed at all concentrations against the Gram-positive S. pyogenes bacteria strain. The chloroform extract has potent antibacterial activity against the Gram-positive S. aureus and Gram-negative P. aeruginosa bacterial strains at a concentration 1mg\1ml with zone inhibition of 10.53mm and 10.83mm. The positive control Beta–sitosterol 0.25% W/W ointment did not show inhibitory effect against four tested bacteria strains (S. aureus, S. pyogenes, Klebsiella, and E. coli) at all concentrations. It has exhibited lesser inhibitory potency when compared with the Jatropha spinosa latex extracts. The present study has revealed that the J. spinosa latex extracts are a potential antimicrobial agent having a broad-spectrum activity.