This study aimed to assess the biometrics of the testes and the morphology of the seminiferous tubules of Crioulo horses. We studied 10 sexually mature stallions (3–6 years of age). After orchiectomy, testes were perfused with Karnovsky's solution and then embedded in glycol methacrylate. Testis sections (4 μm) were cut and stained with toluidine blue and a solution of 1% sodium borate. The histological images were digitized, and the morphometric analysis was performed using ImageJ software. The average weight of the stallions was 377.5 kg, and the average weight of both testicles was 162.9 g. The percentage of testicular parenchyma occupied by the seminiferous tubules and the intertubular tissue was 77.97% ± 6.34% and 22.03% ± 6.34%, respectively. The average tubular diameter was 205.00 ± 36.91 μm, whereas the average height of the seminiferous epithelium was 70.56 ± 2.82 μm. Average tubular length per testicle and average tubular length per gram of testicle were 4,085.10 ± 1,170.68 m and 26.09 ± 10.63 m/g, respectively. The characteristics of the eight stages of the seminiferous epithelium cycle were similar to those reported in other horse breeds. We conclude that the morphometry of the seminiferous tubules of Crioulo horse resembles what has been reported in other horse breeds. The volumetric proportion of the seminiferous tubules and the Leydig cells of the Crioulo horse is one of the highest ever reported for stallions.