The aim of the present study was to investigate the effects of thyme extract (TE) and oxytetracycline (OTC) on rainbow trout (Oncorhynchus mykiss) stress response, immune parameters, oxidative stress and enzymatic changes. The fish were assigned into eight treatments receiving diets with OTC (0 and 2.5 g/kg) and TE (0, 5, 10 and 20 g/kg) for 2 weeks. Thereafter, serum characteristics and gut oxidative status were evaluated. OTC significantly increased serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, and cortisol and glucose levels; however, TE significantly suppressed these changes. OTC significantly decreased, but TE significantly increased gut catalase (CAT) and glutathione peroxidase (GPx) activities. OTC significantly decreased gut superoxide dismutase (SOD) and increased gut glutathione-s-transferase (GST) activities, malondialdehyde (MDA) content and blood monocyte percentage; TE significantly mitigated these changes. TE significantly increased serum total protein, total Ig, lysozyme and ACH50, and blood WBC; however, OTC significantly decreased serum lysozyme and ACH50 activities and blood WBC and neutrophil percentage. In conclusion, OTC administration causes immunosuppression, which might be related to deterioration of fish health. TE augments trout innate immune and antioxidant status and is capable to counteract OTC-induced health and immune deterioration. © 2018 John Wiley & Sons Ltd