Abstract. Liquid culture of seleniferous mycelia of Pleurotus geesteranus Singer (SeMyc-P) was evaluated for its acute toxicity in supplement application and antioxidant against D-galactose induced oxidation damaged mice. Maximum Tolerant Dose (MTD) of SeMyc-P was about 20g/kg bw for mice to define safety application for selenium supplement against acute toxicity. Comparing with carboxymethyl cellulose sodium (CMC) control and VitE standard group, SeMyc-P in 10-days administration increased blood GSH-Px about 24.8-52.0% and 17.1-42.6%, respectivily. By 45-days administration against D-galactose induced oxidative stress mice, the optimal given level of SeMyc-P was 0.2µg/hd/d equivalent to 5-times of DRIs in selenium for Recommended Nutritional Intake (RNI=50μg/d), which increased blood GSH-Px 18.48% than CK-m group concomitant with decreasing MDA and protein carbonyl for 22.19% and 44.16%, respectively.