Abstract: In order to screen Lactobacillus acidophihts with probiotic properties, the intestinal contents samples collected from Weifang and surrounding area of Shandong Province were used to isolate the LactobaciUus strains by conventional method. The isolates with preferable acid production performance were selected by determining the pH value of fermentation liquid. The screened isolates were molecularly identified by using PCR assay and sequencing. The prohiotic properties of the selected isolates were evaluated by conventional method. Among the 9 isolated strains, one isolate with preferable acid production performance, named as XMR-3, was found and was molecularly identified as Lactobacillus acidophilus. The isolate got into the logarithmic growth phase after six-hour incubation. The live bacteria number reached the peak when it was incubated for 16 h, and the pH value of its culture fluid was 3.92. The survival rate of the isolate was above 99% when it was incubated in broth with pH value of 3.0 and 4.0, and that of the isolate was 87.3% when it was incubated in broth with 0.3% bile salt. The antibacterial circle diameters against common pathogenic bacteria were all above 20 ram. The selected Lactobacillus acidophilus isolate showed preferable probiotic properties and bad potentiality in the development of probiotics products for animal use.