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The electrochemical treatment of canal water was investigated in a batch-wise system in the presence of stainless steel 316-grade electrodes. Three effective process parameters, including current density, reaction time, and electrode spacing, were evaluated in the range of 0.25–2.5 mA/cm2, 1–10 min, and 0.5–2.5 cm, respectively. Operational variables of electrochemical disinfection are optimized in response surface methodology (RSM) using Box–Behnken design. Before electrochemical disinfection, a pretreatment process of coagulants mixing for turbidity removal was conducted. Results revealed that a 10 ppm dosage of Ferric chloride (FeCl3.6H2O) and alum (Al2(SO4)3·16H2O) at neutral pH is appropriate. Furthermore, the RSM analysis shows that interelectrode spacing is the most prominent factor affecting the disinfection performance, and increasing electrode spacing inversely affects the disinfection efficiency. Results revealed that 1.52 mA/cm2 current density, 6.35 min reaction time, and 1.13 cm of electrode spacing are the optimum conditions, resulting in a statistically 98.08% disinfection of the total coliform. The energy required for electrochemically disinfection of water at optimum conditions was 0.256 kWh/m3. [ABSTRACT FROM AUTHOR]