In order to know the effects of Garlic, Scallion, Flavonoid, Urushiol, Anthocyanidin and Bio-MOS?? on pathogenic microbes and rumen anaerobic microbes, the growth rate of pathogens (including Escherichia coli O157, Salmonella paratyphi, Listeria monocytogenes and Staphylococcus aureus) and in vitro rumen microbial growth, gas production, ammonia concentration, carboxymethylcellulase(CMCase) activity, and microbial populations were investigated.The growth of pathogens was inhibited by supplementation of 0.1% Flavonoid, Scallion or Bio-MOS?? as biological active materials. And Scallion and Flavonoid had powerful antimicrobial properties on the pathogens applied in paper disc method.Although few effects by biological active materials disappeared in rumen fermentation in vitro, CMCase activity removed with supplementation of 1% of Flavonoid which had antimicrobial property in paper disc method. Scallion, having powerful antimicrobial property on pathogens and no inhibiting on rumen fermentation, might be a source in development of natural antimicrobial agent for ruminants.