[Objective] To evaluate the effect of spent culture supernatant (SCS) of Bifidobacterium on microflora of mice with dysbacteriosis. [Methods] The model of animals with dysbcteriosis was made by means of injecting penicillin into mice adoment, and they were given a therepy of microecology. [Results] There is obvious difference between the normal group and the group of bifidobacterium, and the group of SCS of bifidobacterium. [Conclusions] SCS of bifidobacterium has effect on microflora of mice, which may regulate mice normal physiology functions. It is more obviously helpful to raising bifidobacterium and lactobacillus.