Developing the zoos' ability to assess the reproductive status of the individuals they house is essential to improve the husbandry and management of these species. The use of non-invasive techniques such as fecal hormone analysis has been proven to be a simple and effective way to achieve this. Designed by bioMerieux, mini VIDAS (R) instrument is used in human and veterinary medicine to evaluate different endocrinological parameters, including serum or plasma progesterone. This study evaluates VIDAS (R) Progesterone (PRG) assay's efficacy to monitor fecal progestagens using a simple sample extraction protocol adapted to the zoo environment. We compared (1) VIDAS (R) PRG fecal profiles with established assays specifically designed for fecal progestagens analysis at the VetmedUni (Vienna, Austria) for okapis (Okapia johnstoni), greater one-horned rhinoceros (Rhinoceros unicorns), giraffes (Giraffa camelopardalis reticulata) and hippopotamus (Hippopotamus amphibius) (2) VIDAS (R) PRG fecal profiles with VIDAS (R) PRG serum profiles for African elephants (Loxodonta Africana), giant anteater (Myrmecophaga tridactyla) and white rhinoceros (Ceratotherium simum). Spearman mean correlations were: 0.6748 for African elephants (n = 2 animals), 0.7969 for giant anteater (n = 1 animal), 0.7926 for okapis (n = 2 animals), 0.6072 for greater one-horned rhinoceros (n = 4 animals), 0.6062 for giraffes (n = 4 animals) and 0.5740 for hippopotamus (n = 2 animals). Fecal progestagens analysis revealed estrous cycles in several species: 12.5 +/- 0.5 weeks for African elephants (n = 2 cycles), 15.3 +/- 1.1 days for okapis (n = 6 cycles), 44 +/- 2.1 days for greater one-horned rhinoceros (n = 4 cycles) and 15.5 +/- 0.5 days for giraffes (n = 4 cycles). We observed pregnancies in a giant anteater, an okapi and a hippopotamus. We observed a strong positive Spearman correlation (r> 0.60) for individuals exhibiting estrous cycles. These first results indicate that the mini VIDAS (R) can be used for monitoring of the reproductive status of non-domesticated species and can be a useful tool for the reproductive management through fecal progesterone analysis. A simple extraction protocol was suitable for sample preparation of fecal progesterone metabolite analysis. Further studies using a larger number of individuals per species at different reproductive stages could confirm the relevance of mini VIDAS (R) in the zoo community.