In this study, by-products from alcohol production were examined in terms of their potential application as external carbon sources for enhancing denitrification in biological nutrient removal systems. Three types of batch tests were used to compare the effects of the distillery by-products, such as fusel oil, syrup and reject water, on the non-acclimated activated sludge. Much higher nitrate utilization rates (NURs) were observed for the latter two carbon sources. In the conventional NUR measurements (one-phase experiments), the observed NURs with syrup and reject water were 3.2-3.3 g N/(kg VSS h) compared with 1.0 g N/(kg VSS h) obtained for fusel oils from two different distilleries. When the carbon sources were added at the beginning of the anoxic phase preceded by an anaerobic phase (two-phase experiments), the NURs were 4.2 g N/(kg VSS h) (syrup and reject water) and 2.4-2.7 g N/(kg VSS h) (fusel oils). The heterotrophic yield coefficient, determined based on the conventional OUR measurements, varied in a relatively narrow range (0.72-0.79 g COD/g COD) for all the examined carbon sources. Due to advantageous composition (much higher COD concentrations and COD/N ratios), fusel is a preferred carbon source for practical handling in full-scale wastewater treatment plants.