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® instrument is used in human and veterinary medicine to evaluate different endocrinological parameters, including serum or plasma progesterone. This study evaluates VIDAS® Progesterone (PRG) assay's efficacy to monitor fecal progestagens using a simple sample extraction protocol adapted to the zoo environment. We compared (1) VIDAS® 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 unicornis), giraffes (Giraffa camelopardalis reticulata) and hippopotamus (Hippopotamus amphibius) (2) VIDAS® PRG fecal profiles with VIDAS® 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 ¼ 2 animals), 0.7969 for giant anteater (n ¼ 1 animal), 0.7926 for okapis (n ¼ 2 animals), 0.6072 for greater one-horned rhinoceros (n ¼ 4 animals), 0.6062 for giraffes (n ¼ 4 animals) and 0.5740 for hippopotamus (n ¼ 2 animals). Fecal progestagens analysis revealed estrous cycles in several species: 12.5 ± 0.5 weeks for African elephants (n ¼ 2 cycles), 15.3 ± 1.1 days for okapis (n ¼ 6 cycles), 44 ± 2.1 days for greater one-horned rhinoceros (n ¼ 4 cycles) and 15.5 ± 0.5 days for giraffes (n ¼ 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® 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® in the zoo community.