Objectives The antibacterial effects of traditional Chinese medicine Lianhua-Qingwen formula on two common pathogens in livestock and poultry including Pasteurellamultocida and Erysipelothrix rhusiopathiae were studied to provide reference for its application in veterinary clinic. Methods The minimum bactericidal concentration (MBC) and minimum inhibitory concentration (MIC) of Lianhua-Qingwen formula against three different strains of Pasteurella including Pasteurella X7, Pasteurella YU, Pasteurella SD were measured with the microbroth dilution method. The experiment of drug therapy was conducted. 50 mice aged 4-6 weeks were divided into 5 groups. Each group after infection with Pasteurella was treated with different therapeutic regimens including Lianhua-Qingwen formula, sterile water, amoxicillin, cefaclor for 3 days. The therapeutic effect of Lianhua-Qingw formula on Pasteurella infection was evaluated with the efficacy of treatment. The MIC, MBC, and the efficacy of treatment of Lianhua-Qingwen formula against Erysipelothrix rhusiopathiae were determined according to the anti-Pasteurella test method of Lianhua-Qingwen formula. Results The MIC and MBC of Lianhua-Qingwen formula for different strains of Pasteurella was 4-8 mg/mL and 8-16 mg/mL, respectively. The efficacy of treatment for Pasteurella infection was 60%, significantly higher than that in the control group, and there was no significant difference in treatment compared to the antibiotic oral group. The MIC and MBC of Lianhua-Qingwen formula against SE38 strain of Erysipelothrix rhusiopathiae was 64 mg/mL and 128 mg/mL, and there was no significant therapeutic effect in the mouse infection model. Conclusions Lianhua-Qingwen formula has a good antibacterial effect on Pasteurella, and can be can be further studied as a substitute for antibiotics.