The aim of the present study was to evaluate the efficiency of advanced oxidation processes (electrochemical, Fenton and electro-Fenton) in the removal of oxytetracycline using SS316 and SS316/β-PbO2 anodes. This study was performed experimentally on a laboratory scale in a 250 mL reactor. First, experiments were designed for the electrochemical process using a central composite design, and the optimal conditions for the variables pH(3.53), electric current density(3.85mA/cm2), initial concentration of oxytetracycline (20mg/L) and electrolysis time (42.35min) was obtained; then, under these conditions, the efficiency of Fenton process with FeSO4 variable without the presence of electrodes was evaluated, and its optimal value was 0.3 g/L, and then in the presence of optimal values of the above 5 variables, the efficiency of electro-Fenton process with H2O2 changes were investigated and the optimal value of 0.12 was obtained for H2O2. The removal efficiencies of oxytetracycline in electrochemical, Fenton, and electro-Fenton processes were 84.7%, 73.4%, and 98.2%, respectively. Under optimal conditions, the SS316/β-PbO2 anode electrode enhanced the oxytetracycline efficiency by electron-Fenton process to 100%. The results of bioassay with microorganisms showed that the reduction of toxicity of the effluent treated by electro-Fenton process for Pseudomonas aeruginosa and Staphylococcus aureus was 84.5% and 69%, respectively.