The antibacterial activity of Aloe barbadensis was tested on clinically isolated bacterial pathogens i.e. Enterococcus bovis, Staphylococcus aureus, Escherichia coli, Proteus vulgaris, Proteus mirabilis, Pseudomonas aeruginosa, Morganella morganii, and Klebsiella pneumoniae causing infection in human being. Ethanolic and aqueous extracts were used for the antibacterial effect, which was measured by the appearance of zone of inhibition. Relatively higher MIC concentrations were obtained for gram negative bacteria E. coli and K. pneumoniae, with ethanol extract; however, no inhibitory effect was noted for aqueous extract. Ethanolic extract possesses great inhibitory activity for gram positive bacteria, E. bovis followed by S. aureus. Among gram negative bacteria, highest inhibitory effect was observed with P. aeruginosa, followed by M. morganii, P. mirabilis, and P. vulgaris, which was significant (p < 0.01) than E. coli and K. pneumoniae. Antimicrobial activity tests of crude extract of A. barbadensis were carried out to validate the use of traditional medicinal herbal and results of this study tend to give credence to the common use of A. barbadensis gel and leaf.