BACKGROUND: Dietary supplementation for beef cattle, using natural plant extracts, such as oregano essential oil (OEO), has proven effective in enhancing growth performance, beef production quantity and quality, and ensuring food safety. However, the precise mechanisms underlying these effects remain unclear. This study investigated the impact of OEO on carcass traits, muscle fiber structure, meat quality, oxidative status, flavor compounds, and gene regulatory mechanisms in the longissimus thoracis (LT) muscles of beef cattle. Eighteen steers were randomly assigned to two groups (n = 9 per group) and fed either a control diet (CK) or the same diet supplemented with 20 g of OEO per head per day for 300 days. RESULTS: Oregano essential oil supplementation improved the body weight, carcass weight, meat production, area and diameter of fiber, ether extract, and water-holding power of muscle. Increasing catalase (CAT), peroxidases (POD), glutathione peroxidase (GSH-Px) and by decreasing lipid droplets (LDs) reduced muscle lipid oxidation. However, the color (L*, a*, b*, C* and H°) and the flavor compounds of muscle were affected adversely by OEO. The transcriptome and metabolome indicated the OEO group enriched fat synthesis, proteo-metabolism, antioxidants, and growth significantly. Five key genes (SH2B2, CD209, LOC504773, C1QC, and HMOX) and ten downregulated metabolites (deoxyguanosine, d-melezitose, maltotriose, raffinose, melatonin, quinic acid, orotic acid, hydrocinnamic acid, 2-methylsuccinic acid, and pyridoxal 5'-phosphate) were identified as key biomarkers. These interacted to positively influence the growth, oxidative status, and meat quality of steers positively. CONCLUSION: These findings suggest that OEO, as a natural bioactive compound, can serve as an additive for beef cattle, with a daily dose of 20 g per steer improving growth and meat quality, although it may affect muscle color and flavor negatively. © 2025 Society of Chemical Industry.