This study aimed to describe the normal ultrasonographic image of the gastrointestinal tract and spleen in Egyptian buffaloes (Bubalus bubalis). The obtained results were compared to the normal reference values that had been previously established in cattle. This study was carried out on thirty healthy buffalo. Complete clinical examination, hematological pictures profiling, blood biochemical analysis of different parameters such as total blood serum proteins, albumins, globulins, aspartate aminotransferase, alkaline phosphatase, gamma-glutamyl transferase, triglycerides and cholesterol, radiography and ultrasonographic examination were done. The clinical findings, blood pictures and blood biochemical levels agreed with the previously reported reference values of healthy buffaloes. Radiography of the healthy buffaloes showed free reticulum, normal heart and well-identified diaphragm. The heart appeared as radio-opaque structure. This study could detect the suitable site for visualization of reticulum, omasum, abomasums, rumen, small intestine and large intestine. It also described the normal ultrasonographic patterns of these organs. It was very essential to establish these ultrasonographic findings to improve the diagnostic efficacy of ultrasonography for evaluation of different gastrointestinal organs in healthy buffaloes that will be very useful in the future detection of the abnormal findings associated with gastrointestinal diseases.