The stimulatory role of equine Chorionic Gonadotropin (eCG) in the production of steroid hormones was evaluated during the first 4 months of pregnancy in mares impregnated by either stallions or jack donkeys. Twenty mares were divided in two groups: Mares in the first group were inseminated with stallion semen (horse pregnancies), and those in the second group were inseminated with donkey semen (mule pregnancies). Blood samples were collected twice weekly from day 30 to day 120 of pregnancy to determine the concentrations of eCG, progesterone, androstenedione, and testosterone. Analysis of variance for repeated measures was used to compare the concentrations of each hormone between groups. Linear regression models that considered the linear and quadratic effects of week of gestation as well as the linear and quadratic effects of the concentrations of eCG on the production of each steroid hormone were carried out. Concentrations of eCG, progesterone, and androstenedione were higher in horse than in mule pregnancies (P < .01 for eCG and P < .05 for progesterone and androstenedione). Testosterone concentrations were also higher in horse pregnancies than in mule pregnancies at weeks 7, 9, and 10 (P < .05). Regression analysis indicated that eCG had considerable stimulatory effects on the secretion of progesterone and androstenedione and weaker effects on the secretion of testosterone. The results suggest that eCG stimulates luteal production of progesterone, androstenedione, and testosterone in horse and mule pregnancies, these effects being more evident in horse pregnancies than in mule pregnancies due to the higher concentrations of eCG in horse pregnancies.