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Objectives of the study were to measure water hardness in Burdur, and to establish its possible association with urolithiasis in cattle. Water samples were obtained from different stables (n = 15). Water hardness and the concentrations of potassium, calcium, magnesium, sodium, iron, zinc, manganese and copper ions were calculated from these water samples. Total hardness of the samples (mean 285 ppm) exceeded the standards and the water was characterized by high content of magnesium ions. Kidneys (n = 500) were collected randomly from slaughterhouses and examined for urolithiasis. Urolithiasis was observed in 102 kidneys (20.4%). The weights of the stones were between 0.02 and 237.44 g and the colour varied from white to brown. The calculi collected had various shapes and composed of calcium apatite (42.45%), struvite (20.15%), magnesium carbonate (15.15%), calcium carbonate (12.12%), and calcium phosphate cystine (10.13%). It was concluded that high water hardness with high magnesium ion concentrations in water may contribute to urolithiasis and needs to be investigated further in future studies.