Obstructive urolithiasis is a serious problem that negatively affects the quality of life in buffalo calves. The present study aims to evaluate the relationship between oxidative stress, antioxidant enzyme activity, and obstructive urolithiasis in buffalo calves and tube cystostomy as a surgical technique to treat such conditions. Seventy-seven non-castrated male buffalo calves aged 3-10 months were admitted to the Faculty of Veterinary Medicine clinic, Zagazig University, Egypt, with a history of anuria. Based on clinical, ultrasonographic, and biochemical examination, the calves were diagnosed with either intact bladder (n=32 calves) or ruptured bladder (n=45 calves). Ten healthy buffalo calves of the same age were used as a control for a reference range of biochemical analysis. Serum levels of malondialdehyde (MDA) significantly increased in all affected calves, especially those with ruptured bladders. In addition, a significant decrease in glutathione peroxidase (GSH) was reported in all affected calves, with lower values in calves with ruptured bladders. The tube cystostomy technique succeeded in 95.95% of the operated cases, and only 4.05% had complications. In conclusion, there is a relation between oxidative stress, antioxidant activity, and obstructive urolithiasis in buffalo calves with useful management using the tube cystostomy technique.