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Purpose: It is well-recognized that dietary factors may influence in the pathogenesis of urinary lithiasis (UL), the third most prevalent disorder of the urinary tract. Herein we evaluated the effects of the dietary supplementation with vitamin D, which has been recently associated with hypercalcemia, on the occurrence of UL in rats that developed chronic hyperoxaluria after exposition to ethylene glycol, an inducing agent. Materials and method: Thirty Sprague-Dawley male rats were randomly divided into three groups: Group 1 (control, n=10); Group 2 (0.5% ethylene glycol+vitamin D3, n=10); Group 3 (1.25% ethylene glycol, n=10). Urine samples were collected over a 24 h period at the baseline (day zero) and weekly during four weeks for the dosage of calcium, oxalate, uric acid, citrate and serum creatinine. Animals were euthanized, their right kidneys removed and the corresponding hematoxylin-eosin staining of the histological sections subjected to histological/ histomorphometric analyses using the Image J® software. The deposition of calcium salts in the renal parenchyma was quantified by the PIXE technique (Proton Induced X-Ray Emission). Results: All animals displayed normal levels of serum creatinine (median 0.6 mg/dl) and no statistical difference was found in the daily fluid intake. The volume of the urine samples was significantly higher in animals from G2 when compared to the control animals (10.1 mL versus 4.5 mL, p