Antibiotic resistance is being constantly developed worldwide. Coagulase Negative Staphylococci (CNS) and Staphylococcus aureus are common causes of bovine subclinical mastitis. Bioactive compound of medicinal plants shows anti-microbial, anti-mutagenic and anti-oxidant effects. The anti-bacterial and anti-oxidant activities of Liquidambar orientalis (L. orientalis) extracts on subclinical mastitis causing bacteria in cows have not been reported to date. The aim of the present study was to examine anti-bacterial and anti-oxidant effects of L. orientalis leaf extracts on S. aureus and CNS isolated from cows with subclinical mastitis symptoms. In this study, 3.2 mg/mL minimum inhibitory concentration (MIC) of ethanol extracts of L. orientalis has shown to be a most potent anti-bacterial and anti-oxidant for all isolated bacterial species from mastitis cows. In this study, it was investigated anti-bacterial and anti-oxidant potentials of acetone, methanol and ethanol extracts of the L. orientalis. The acetone extract showed maximum inhibition zone against S. aureus numbered 17 (12 mm). In addition to anti-bacterial properties, anti-oxidant activity of L. orientalis extract was examined by ABTS [2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid)] free radical assay. Trolox was used as a positive control anti-oxidant. Ethanol extract exhibited a strong anti-oxidant activity like Trolox anti-oxidant which was effective at 2.58 mM concentration. Bioactive compounds of sweet gum may be useful to screening mastitis causing bacteria for clinical applications.