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Probiotics are live microorganisms that confer useful health effects on the body of human beings when provided in sufficient amounts. The probiotics isolated from natural foods have demonstrated the ability to inhibit the microbial growth and reduce cholesterol levels in the human body. Nevertheless, scanty research work on the activity of probiotics especially the lactobacillus strain obtained from the bovine tripe has so far been carried out. Natural foods are more preferable and acceptable in the modern society than pharmaceutically prepared food products and therefore this study was conducted to isolate probiotic organisms from a natural food for health benefits. This study also focused on obtaining the Lactobacillus strain to be consumed directly as a diet. The cultures of Lactobacillus strains were isolated and then characterised morphologically and biochemically at the species level. Results showed that isolated strain (Lactobacillus acidophilus) had the ability to remain alive and proliferate at pH 3.0 -7.0 with optimum growth pH at 4.0. In addition, all isolates tolerated 0.0, 0.1, 0.3, 0.5, and 1.0 % bile salt concentrations. The test antibiotics sensitivity tests showed that the strains could resist streptomycin, Neomycin, penicillin and Vancomycin. However, the strains were sensitive to Erythromycin, Azithral and Chloramphenicol. The strain also reduced serum cholesterol level as far as 67.0 % which is a crucial finding. Therefore, bovine tripe is a potential source of probiotics that can be used in the development of new products that can benefit human health.