This study comparatively investigated the exoelectrogenic utilization and hydrogen conversion of major dark fermentation products (acetate, propionate, butyrate, lactate, and ethanol) from organic wastes in dual-chamber microbial electrolysis cells (MECs) alongside their mixture as a simulated dark fermentation effluent (DFE). Acetate-fed MECs showed the highest hydrogen yield (1,465 mL/g chemical oxygen demand), near the theoretical maximum yield, with the highest coulombic efficiency (105%) and maximum current density (7.9 A/m2), followed by lactate-fed, propionate-fed, butyrate-fed, mixture-fed, and ethanol-fed MECs. Meanwhile, the highest hydrogen production rate (514 mL/L anolyte center dot d) was observed in ethanol-fed MECs despite their lower coulombic efficiency. Butyrate was the least favored substrate, followed by propionate, leading to significantly delayed startup and reaction. The active anodic microbial community structure varied considerably among the MECs utilizing different substrates, particularly between Geobacter and Acetobacterium dominance. The results highlight the substantial effect of the DFE composition on its utilization and current-producing bioanode development.