The performance of hydrological models is affected by uncertainty related to observed climatological and discharge data. Although the latter has been widely investigated, the effects on hydrological models from different starting times of the day have received little interest. In this study, observational data from one tropical basin were used to investigate the effects on a typical bucket-type hydrological model, the HBV, when the definitions of the climatological and discharge days are changed. An optimization procedure based on a genetic algorithm was used to assess the effects on model performance. Nash-Sutcliffe efficiencies varied considerably between day definitions, with the largest dependence on the climatological-day definition. The variation was likely caused by how storm water was assigned to one or two daily rainfall values depending on the definition of the climatological day. Hydrological models are unlikely to predict high flows accurately if rainfall intensities are reduced because of the day definition.