The invention discloses a prescription of a compound berberine sulfate injection for veterinary use and a preparation process thereof, belonging to veterinary drug technologies. The prescription consists of a main drug, an effect extending agent, a cosolvent, an antioxidant and water, and a high-concentration injection can be prepared. The injection is advanced in process, high in stability, long-acting, broad in spectrum, high-efficiency, small in irritation to the injection part and convenient to use, and the curative effect of the injection is obviously superior to that of single preparations of a berberine sulfate injection and an enrofloxacin injection. The injection comprises the following raw materials: 0.5-5.0 percent of berberine sulfate, 2.5-20.0 percent of enrofloxacin, 1-10 percent of effect extending agent, 1-10 percent of cosolvent, and 0.1-0.5 percent of antioxidant, wherein the effect extending agent refers to one or a mixture of several in polyethylene glycol, polyvinyl alcohol, povidone, hydroxyethyl cellulose, hydroxy propyl cellulose, carboxymethyl cellulose, hyaluronic acid, xanthan gum, gelatin, chitosan and alginate, the cosolvent refers to one or a mixture of several in sulfuric acid, hydrochloric acid, lactic acid, citric acid and acetic acid, and the antioxidant refers to one or a mixture of several in sodium hydrogen sulfite, anhydrous sodium sulfite, sodium formaldehyde sulfoxylate, sodium pyrosulfite, thiourea, L-cysteamine hydrochloride, vitamin C and ethidene diamine.