The Erythrinidae trahira, Hoplias malabaricus (Bloch, 1794), is widespread throughout South America river basins. We determined Sertoli cell supporting capacity (ratio of primary spermatocytes: Sertoli cells and spermatids: Sertoli cells), meiotic index (ratio of spermatids: primary spermatocytes) and the number of spermatogonial mitotic generations of this fish. The fish were captured in the Igarapava reservoir, Grande River, Alto Paraná River basin, Brazil. Testis fragments of three sexually mature trahiras were fixed in 5% buffered glutaraldehyde solution and embedded in glycol methacrylate. Serial sections of 2 and 3 µm in thickness were stained with 0.5% toluidine blue. Histological counts from cysts of primary spermatocytes and spermatids revealed, respectively, 326 ± 99 and 468 ± 73 nuclei of these cells. Sertoli cell supporting capacity was considerably higher for spermatids (113.3 ± 16:1) when compared to primary spermatocytes (71 ± 5:1). Between eight and ten spermatogonial generations were formed to give rise to primary spermatocytes. These values were within the generation range of those already found in freshwater teleosts of external fertilization. Correlation between the number of Sertoli cells and primary spermatocytes per cyst, and Sertoli cells and spermatids per cyst were statistically significant (p < 0.05).