This study aimed to validate the enzyme immunoassay (EIA) for fecal progestin quantification of the species Mazama americana, define its excretion profile during periods of gestation and postpartum and determine the gestation period and resumption of postpartum ovarian activity in this species in captivity. Fecal samples were collected twice a week during gestation and every day in the postpartum period, and analyzed using EIA. The mean concentrations (±SEM) of fecal progestins during gestation were 2180.0±299.1ng/g in early pregnancy (week 1-11), 3271.4±406.9ng/g in middle pregnancy (week 12-22) and 5592.0±1125.8ng/g in late pregnancy (week 23-32). The gestation period determined for the species was 220.9±1.2 days. The concentration of progestins reached its peak prior to parturition and returned to baseline levels in 4±0.31 days after parturition. In the postpartum period, the mean concentrations of fecal progestins were 1564.2±182.6ng/g in the interval between parturition and resumption of ovarian activity, 469.8±24.5ng/g in the inter-luteal phase and 2401.7±318.5ng/g during the luteal phase, such that the postpartum period and the luteal phase differed from the inter-luteal phase. Fecal progestin profiling permitted the detection of ovulation 26.9±3.4 days after parturition in all the hinds studied and estimation of the mean duration of the estrous cycle, 21.3±1.1 days. Analysis established that concentrations of progestins above 3038.76ng/g diagnosed pregnancy, a value determined from the week 12 of gestation. Moreover, the quantification of fecal progestins by EIA proved to be an important tool for noninvasive endocrine monitoring and to obtain reproductive data on the species M. americana in captivity. Copyright © 2013 Elsevier B.V. All rights reserved.