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Abstract Objective: This study aims to evaluate the effect of electrolyzed water effect on bacterial colonization on diabetic foot ulcer (DFU) patients in Makassar. Method: This is an in vitro. Eight bacteria are incubated into 5 types of electrolyzed water (ESAW), electrolyzed weak acid water (EWAW), electrolyzed neutral water (ENAW), electrolyzed weak alkaline water (EWAlW), and electrolyzed strong alkaline water (ESAlW). Evaluations were performed 2, 24, 48 and 72 hours after incubation. Data were analyzed using repeated ANOVA test used to compare the difference of electrolyzed water effect on bacterial colonization of DFU patients. Results: ESAW (pH 2.5) significantly (p = 0.001) had a better bactericidal effect than EWAW, ENAW, EWAlW and ESAlW. Conclusion: There is a difference of electrolyzed water effect on bacterial colonization in DFU patients.