Color removal was evaluated in anaerobic assays using synthetic (SE) and real (RE) effluents containing the dye Direct Black 22 (DB22). The focus was on the individual influence of two electron donors (EDs), sucrose and ethanol; and their combination with two redox mediators (RMs), lawsone and riboflavin. In addition to SE assays, the EDs were combined with sulfate which was present in the RE. Distinct behavior was observed with both assays. In the case of SE, using the EDs sucrose and ethanol resulted in DB22 removal efficiency of 93 and 83%, respectively; but only 27% in the control. The respective removal rates were 27.8‐ and 4.4‐fold higher than that of the control. Using the EDs with the RM lawsone resulted in color removal higher than that with riboflavin. Interestingly, in the ethanol + sulfate assay, late release of the last was observed, possibly due to the desulfonation reaction of the aromatic structures during the DB22 molecule breakdown. In the case of RE, no improvement was observed with either EDs; and some combinations with the RMs even resulted in decreased color removal. Therefore, the used EDs and RMs showed positive effect on color removal in SEs; but unexpectedly not in REs.

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