The fate of N species, particularly dissolved organic nitrogen (DON), through process trains of a wastewater treatment plant (WWTP) was investigated. In this study, three fully nitrifying plants in Illinois, USA and biological nutrient removal (BNR) plants in northern Poland were sampled for N characterization in the primary and secondary effluents as a function of the particle size distribution. The correlations between dissolved organic carbon (DOC) and dissolved organic nitrogen (DON) concentrations were examined. The key findings are that DON becomes significant portion (about 20%) of the effluent N, reaching up to 50% of effluent total N in one of the Polish plants. The DON constituted 56–95% of total ON (TON) in the secondary effluents, whereas in the Polish plants the DON contribution was substantially lower (19–62%) and in one case (Gdansk WWTP) colloidal ON was the dominating fraction (62% of TON). The DOC to DON ratio in the US plants is significantly lower than that in the receiving waters indicating potential for deterioration of receiving water quality. In Polish plants, the influent and effluent C:N ratios are similar, but not in the US plants.