Abstract: In order to investigate the biological properties of Bacillus licheniformis from different sources, such as the inhibition effect on common pathogenic bacteria, the experiment was performed by using bacterial morphology, physiological and biochemical test, 16S rRNA gene sequencing, phylogenetic tree analysis, Oxford Cup inhibition test and other detection techniques; five strains of Bacillus licheniformis from different sources（chicken manure, sheep manure, cattle rumen contents, pig intestinal contents and soil） were isolated, identified and detected for in vitro inhibition activity. The results showed that five strains B₁₇（chicken manure）, B₃₀（sheep manure）, B₃₂（bovine rumen contents）, B₃₅（pig intestinal contents） and B₃₈（soil） could grow on LB solid medium, with three strains from different sources, B₃₀, B₃₅ and B₃₈, having the same colony morphology. The microscopic examination showed that the strains were all Gram-positive bacillus with subcentral spores, which were consistent with the characteristics of Bacillus; the physiological and biochemical characteristics were consistent with those of Bacillus; the PCR amplification yielded a target fragment of about 1 500 bp. All strains were in the same cluster with Bacillus licheniformis in GenBank and were identified as Bacillus licheniformis. Among the five strains of Bacillus licheniformis, except for pig-derived Bacillus licheniformis B₃₅, the other four strains had bacteriostatic effect, and sheep-and cattle-derived Bacillus licheniformis B₃₀ and B₃₂ had better bacteriostatic effect and a wider range of bacteriostasis. The results suggested that Bacillus licheniformis isolated from sheep manure and bovine rumen contents could be used as a probiotic Bacillus to control pathogenic bacteria, and different sources of Bacillus licheniformis had different bacteriostatic effect.