This study examined the effects of stevioside (S) and organic selenium (O-Se) supplementation on the sensory attributes, microbial activity, fatty acid composition, and meat quality traits of Hanwoo cattle (Korean native cattle). Twenty-four Hanwoo cattle (663 ± 22 kg body weight) were assigned to two dietary treatments for 8 months: control diet and 1% stevioside with 0.08% organic selenium supplemented diet. S and O-Se inclusion in the diet enhanced the final body weight, weight gain, and carcass crude protein (p < 0.05). Moreover, supplementation with S and O-Se had a significant effect on lowering the drip loss and shear force and enhanced the a* (redness) of the longissimus dorsi muscle (p < 0.05). The inclusion of dietary S and O-Se improved the sum of the polyunsaturated fatty acid (ΣPUFAs) content of the meat, and the oxidative status (TBARS) values during second week of storage decreased by 42% (p < 0.05). On the other hand, the microbial count tended to decrease (7.62 vs. 7.41 log10 CFU), but it was not significant (p > 0.05), and all sensory attributes were enhanced in the S and O-Se supplemented diet. Overall, these results suggest that supplementation of the ruminant diet with stevioside and organic selenium improves the growth performance, carcass traits, and meat quality with enriched PUFAs profile and retards the lipid oxidation during the storage period in beef.