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A passive treatment process using sulfate-reducing bacteria (SRB) is known to be effective in removing heavy metals from acid mine drainage (AMD), though there has been little discussion of the mechanism involved to date. In this work, a sulfate-reducing column test was carried out using supplementary ethanol as an electron donor for microorganisms, and the reaction mechanism was examined using geochemical modeling and X-ray absorption fine structure (XAFS) analysis. The results showed that Cu was readily removed from the AMD on the top surface of the column (0–0.2 m), while Zn and Cd depletion was initiated in the middle of the column (0.2–0.4 m), where sulfide formation by SRB became noticeable. Calculations by a developed geochemical model suggested that ethanol decomposition by aerobic microbes contributed to the reduction of Cu, while sulfide produced by SRB was the major cause of Zn and Cd removal. XAFS analysis of column residue detected ZnS, ZnSO4 (ZnS oxidized by atmospheric exposure during the drying process), and CuCO3, thus confirming the validity of the developed geochemical model. Based on these results, the application of the constructed geochemical model to AMD treatment with SRB could be a useful approach in predicting the behavior of heavy metal removal. [ABSTRACT FROM AUTHOR]