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The objective of this study was to assess the antibacterial activity of Rifampicin-loaded solid lipid nanoparticles on Brucella abortus 544. Rifampicin-loaded solid lipid nanoparticles were prepared by a modified microemulsion/sonication method and characterized. The results showed the average size about 319.7 nm, PI about 0.20 and zeta potential about 18.4 mv, encapsulation efficacy and drug-loading were equal to 95.78 and 34.2%, respectively, with a spherical shape. Drug release lasted for 5 days. The antibacterial activity was statistically significant with p < .05 in bacterial and cell culture media compared to free Rifampicin. It can be concluded that solid lipid nanoparticles can be considered as a promising delivery system for improving the antibacterial activity of Rifampicin against Brucella abortus.