The invention discloses a preparation method of a clostridium butyricum compound bacterial solution for livestock and poultry. The preparation method comprises the steps as follows: A, producing a compound flora material; B, producing a first-class liquid strain: (1) preparing a culture solution; (2) putting the prepared culture solution in a conical flask, performing sealing and sterilization, then, cooling the culture solution to 25 DEG C or below, inoculating the culture solution with parent mycelia blocks obtained in step A, inoculating every 100 ml of culture solution with 1-2 parent mycelia blocks 2 mm long and wide respectively; adding 10-50 ml of clostridium butyricum activated propagation solution obtained in step A; uniformly mixing strains, and then, performing culture for 1-2.5days to obtain a first-class liquid strain; and C, producing a second-class liquid strain to obtain a compound liquid probiotic agent. Research topic results are taken as the basis, the prepared compound bacterial solution contains specific probiotic florae, and improvement of sanitary environments of livestock and poultry and control of diseases can be carried out by spraying the compound liquidprobiotic agent on livestock and poultry bodies.