According to the Ukrainian official statistic reporting data, the total number of registered water users within the Ukrainian part of the Dnipro Basin is about seven thousand, with 903 effluent discharge outfalls being located along the rivers draining the Basin. The following criteria were used for identification of Hot Spots and preliminary screening of industrial enterprises and wastewater treatment utilities that discharge their effluents directly into the rivers of the Dnipro Basin: (1) discharge intensity and type; and (2) discharged effluent composition/quality, treatment and monitoring. In the process of identification and screening of pollution sources, preference was given to those sources that discharge large volume of effluents containing specific substances at higher concentrations, and thereby affect water quality in a river. Special focus was placed on those industries that discharge their effluent without any treatment, and/or provide only partial treatment. It should be noted that the existing monitoring of discharges is severely impaired by the lack of adequate funding, which can be attributed to a difficult economic situation in the country as a whole. In many cases, it is difficult to evaluate and assess the monitoring results with any confidence. Therefore, it was considered useful to expand the hot spot assessment methodology by a component that enables additional characterization of water quality downstream of effluent discharges (using data collected at the hydrometricological monitoring stations) in terms of key parameters. This approach would provide a greater degree of objectivity at a very early stage of the Hot Spot selection process.