Collected from the broiler cecal contents, bacteria were isolated and purified. These isolated and purified Lactobacilli were detected by acidity and bile salt tolerant test, then three better tolerant strains were screened out. These three strains were conducted to observe the morphous through microscope, perform Gram stain, and to identify their physiological and biochemical characteristics. The results showed that L1 was L.casei, L2 was L.gasseri and L3 was L.graminis. With Oxford Cup mcthod, these three Lactobacilli were detected for their bacteriostasis. Results showed that L1, L2 and L3 strains had strong antibacterial activities to E. coli and Salmonella.