Phenotypic, andrologic, and cytogenetic investigations were performed on a 2.5 years old buffalo-bull that produced azoospermic semen from puberty. Also, ultrasonographic and histopathologic examinations were conducted on its testis. Phenotypic examinations showed atypical male features. Andrological investigations revealed small non-pendulous scrotum and reduced testicular size as indicated by lowered scrotal circumference measurement and ultrasound-detected testicular breadth. Both libido and serum testosterone concentration were greatly reduced. The sonographic examination of the test is revealed hypoechoic feature and reduced testicular breadth. Testicular hypoplasia was confirmed by the histopathological findings represented by a marked reduction in the number and diameter of seminiferous tubules and absence of mitotic or meiotic activities. Cytogenetic examination of peripheral leucocytes revealed the presence of trisomy-24.It could be concluded that identifying trisomy -24 as possible cause of testicular hypoplasia in buffalo bulls is being useful for earlier diagnosis of the affected cases before reaching the age of puberty when the condition could be diagnosed and thus reducing the costs of keeping them till puberty.