Anaerobic reactors were operated in a semi-batch mode and fed with the dual substrates glucose (G) plus acetic acid (Ac) as primary organic sources to study the effect of sulfate on COD oxidation. With glucose, COD removal by methane formation was seriously inhibited, resulting in COD accumulation in the reactor. Although acetic acid can be consumed by some sulfate-reducing species, it was not a major substrate for sulfate reduction, but was largely responsible for methane formation in the anaerobic mixed culture used in this study. With dual substrates, extreme inhibition of methanogenesis did not occur as did with glucose alone. Instead, methanogens were found to work in harmony with acid formers as well as sulfate reducers to oxidise COD. Interestingly, from 12-hour monitoring, increased G/Ac COD ratios decreased COD removal rates as well as biogas production, but resulted in higher sulfate reduction. This suggests that there should be an optimal feed G/Ac COD ratio, for which removal of both organics could be maximised.