Abstract: The aim of the present study was to evaluate the antibacterial effect of single and combined application of traditional Chinese herbs against Staphylococcus isolates from poultry in vitro. The minimal inhibitory concentration （MIC） of thirteen kinds of traditional Chinese herbs against clinical isolates of poultry origin Staphylococci was determined by tube double dilution method. Five kinds of traditional Chinese herbs with better antibacterial activity were screened, and the fractional inhibitory concentration index （FIC index ） against Staphylococci when the five kinds of traditional Chinese herbs were combinedly used was determined by checkerboard dilution method. The results showed that the aqueous extract of uncaria and dark plum showed the best antibacterial activity in vitro against the poultry origin Staphylococci and both of their MIC was less than 1.95 mg/mL, followed by that of coptis and sappan wood with MIC of 3.91 mg/mL. The synergistic antibacterial effect was observed in the combined application of honeysuckle and myrobalan, and the other combinations exhibited irrelevant or antagonistic action.