Abstract: To screen Lactobacillus with strong ability of selenium resistance and selenium enrichment, the selenium resistance of Streptococius thermopiles, Lactobacillus bulgaria, and Lactobacillus acidophilus is compared, and the selenium-enriched study is conducted on the strain with the strongest selenium resistance. The separation and purification, combined with glycolytic and Gram staining, was used to identify the strains, and then the strains were domesticated (the media should form the concentration differences). The 3,3''-aminobenzidine colorimetry was used to determine the selenium conversion. The results showed that the order of selenium tolerance was Lactobacillus acidophilus, Streptococius thermopiles and Lactobacillus bulgaria. When the selenium content was 17 μg/mL, the strains were cultivated for 9 h under 37℃, the selenium enrichment rate of Lactobacillus acidophilus reached 65.4%. Lactobacillus acidophilus colonies were generally found to be relatively small, irregularly rounded, milky white, smooth and shiny on selenium-containing media