Interior microelectrolysis (IM) as a pretreatment process was effective to treat Reactive Black 5 (RB5) in this study. The removal rates of chemical oxygen demand (COD), total organic carbon (TOC) and color were 46.05, 39.99 and 98.77%, respectively, when this process was conducted under the following optimal conditions: the volumetric ratio between iron scraps and active carbon (AC) (V(Fe)/V(C)) 1.0, pH 2.0, aeration dosage 0.6 L/min, and reaction time 100 min. Contaminants could be further removed by ozonation. After subsequent ozonation for 200 min, the solution could be completely decolorized, and the COD and TOC removal rates were up to 77.78 and 66.51%, respectively. In addition, acute toxicity tests with Daphnia magna showed that pretreatment by IM generated effluents that were more toxic when compared with the initial wastewater, and the toxicity was reduced after subsequent ozonation.