Olive flounder was treated with oxytetracycline (OTC) and changes in blood physiology, antioxidant enzymes and heat shock protein (HSP) were recorded to obtain preliminary data for optimal OTC treatment. Blood parameters were measured 1 and 3 h after the OTC treatments at the concentration of 0 (control), 100, 300 and 500 ppm for I h. Hematocrit decreased with time, however the difference was not significant (P>0.05). Reduced number of red blood cell was observed with increasing OTC concentration. Serum glucose level increased as the OTC concentration increased. However, glucose level was similar to control after 3 h. Blood total protein decreased immediately after the OTC treatment but increased after 1 and 3 h. However, the increment in blood total protein was low. Activities of superoxide dismutase enzymes in 300 and 500 ppm groups increased by the OTC concentration. Catalase enzyme activity was negatively affected by the OTC concentration. However, the differences were not significant (P>0.05). High expression of HSP-70 protein was recorded for groups treated with 100 and 500 ppm compared to that of the control group. However HSP-70 mRNA showed a lower increment which was not significant (P>0.05).