Background and Aim: Obstructive urolithiasis is one of the major health problems in livestock animals, mainly in young calves. The present study was designed first to investigate the changes in the serum levels of Vitamins A and D in buffalo calves (Bubalus bubalis) with obstructive urolithiasis and second to investigate the efficacy of tube cystostomy technique in management of such condition. Materials and Methods: One hundred and forty-nine buffalo calves of variable ages ranged from 3 to 7 months with a history of retained urine were examined clinically and ultrasonographically. Then, they were subjected to surgical treatment using the tube cystostomy technique. The serum levels of Vitamins A and D were investigated in retained urine calves in addition to 10 clinically healthy calves of the same age used as a control group. Results: Based on clinical and ultrasonographic findings, the calves were diagnosed as obstructive urolithiasis with intact bladder (n=64 calves) or with bladder rupture (n=85 calves) with the peak incidence in winter months. Tube cystostomy was an efficient and quick surgical technique for the management of such condition and 95.3% of calves returned their normal urination within 7–14 days after surgery. Significant hypovitaminoses A and D were found between retained urine calves and control ones (p= 0.01 and 0.002, respectively). Conclusion: Hypovitaminoses A and D suggested predisposing obstructive urolithiasis in buffalo calves, but further clinical studies are recommended for more confirmation. Surgical treatment using tube cystostomy technique is recommended for the management of obstructive urolithiasis in buffalo calves.