This article is dedicated to evaluation of water use in the Botna river basin – one of the biggest tributary of the Dniester River in the limits of the Republic of Moldova. Botna river basin occupies a total surface of 1518 km3 and includes areas of 5 administrative districts from the central and south-eastern part of the Republic. In the study are situated over 60 settlements, including only 2 medium and small towns (Căușeni and Căinari). The study is based on the approaches and methodology proposed by the EU Water Framework Directive and its methodological guidelines on the economic assessment of water use and on the national normative documents in the field and will be used in the elaboration od the Management Plan of Botna river basin. In the years 2010-2018, the total volume of water used in the Botna river basin was, on average, 3.1 million cubic meters, of which 85% were captured from underground sources. Over 50% of the total volume of water was used for household purposes, about 1/3 - for agricultural needs and only 13% for industrial activities. Also, are registering an oscillating dynamics of total volume of used water, depending on: the capacity of the water pumping stations; transport capacity of public aqueducts; local water resources; the annual amount of rainfall; the economic activity of big agricultural companies and its capacity to records of abstracted an used water. As a result of rapid extension of public aqueducts in the rural area, is observed an increase more than 2 times the volume of water used for household purposes. In the same time, it is registered a ≈4 times reduction of the volume of water used by agricultural companies. Also, there is a slow reduction of the water used in the industry. The key problems of water use in the research area are: intensification of climate change and reduction of water supply access of households and agriculture companies; incomplete inventory of water resources and frequent cases of unauthorised water use; low prices of water supply services reduced accessibility of the population to aqueducts; significant impact of diffuse pollution on agriculture; a large number of reservoirs in improper conditions, which do not cover water use costs and investments in water resources and aquatic ecosystems protection; insufficient and fragmented subventions in water sector.