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Phosphorus belongs to the crucial bioelements that cause eutrophication, and phosphates, easily assimilated by organisms, are widespread in the environment. Phosphates can be of natural or anthropogenic origin and can derive from various point or non-point sources. Knowledge about the origin of nutrients is necessary to effectively manage, protect, and revitalize water resources. To recognize various phosphate sources in the study area of our research, i.e., the Vistula and Bug interfluve (SE Poland), we used the oxygen isotopic signature of phosphate ions (δ18O-PO4), which has been successfully used in recent decades as a tracer of phosphorus cycling in water studies. We measured the δ18O-PO4 of dissolved inorganic phosphates (DIPs) extracted from various phosphate sources. The obtained results are as follows: For springs, the δ18O-PO4 value varied from +14.8‰ to +18.5‰; for riverine samples, from +10.3‰ to +18.6‰, which were significantly location-dependent; while waste water treatment plant effluents ranged from +12.4‰ to +15.6‰. Two tested drainage water samples had similar isotopic compositions (+16.7‰ and +17.3‰). In the case of two analyzed bedrock samples, the δ18O-PO4 values, which were similar (+20.5‰ and +21.7‰), are close to the existing data on sedimentary bedrocks derived from similar geological periods. The obtained results can be helpful in future research aimed at identifying phosphate sources and P cycling in the studied area. [ABSTRACT FROM AUTHOR]