Ghrelin, the endogenous ligand for the GH-secretagogue receptor (GHS-R), has been found in the testes of many vertebrates, and in the cerebellum and gastrointestinal tract of African ostrich, but there is little known about its distribution in the testes of the African ostrich. In the present study, the distribution and morphological characteristics of ghrelin-immunopositive (ghrelin-ip) cells in the testes of African ostrich were investigated using immunohistochemistry. The testes of African ostrich consisted of a capsule and parenchyma. The capsule was divided into three annular layers, and the parenchyma consisted of tubular compartments and interstitial tissue. The tubular compartments were comprised of the seminiferous tubules, straight seminiferous tubules and rete testis, without any distinct boundary tissue between them. The interstitial tissue between the tubules contained many Leydig cells, tiny veins and connective tissue. Ghrelin-ip cells in the African ostrich testes were localized only in the parenchyma, and mostly in the tubular compartments. The ghrelin-ip cells in the tubular compartments were mainly spermatogenic cells and some Sertoli’s cells. In the insterstitial tissue, the ghrelin-ip cells were identified as Leydig cells. The ghrelin-ip cells showed cytoplasmic staining, and were irregularly shaped. These findings indicate that ghrelin may participate in the spermatogenesis in the African ostrich. Further studies into this regulation will help clarify the physiological roles played by this hormone in the African ostrich.