Edwards, KatieL.Wheaton, Catharine J.Brown, Janine L.Dimovski, Alicia M.Fanson, Kerry V.Ganswindt, AndreGanswindt, Stefanie BirgitHagenah, NicoleKeeley6, TamaraMöstl, ErichO’Hara, BobbiPenfold, LindaM.Shablin, Samantha A.Palme, Rupert2025-12-032025-12-032025-10Edwards, K.L., Wheaton, C.J., Brown, J.L., Dimovski, A.M., Fanson, K.V., Ganswindt, A., Ganswindt, S.B., Hagenah, N., Keeley, T., Möstl, E., O’Hara, B., Penfold, L.M., Shablin, S.A. & Palme, R. (2025) Developmentofan 11-oxoetiocholanolone mini-kit for the quantification of faecal glucocorticoid metabolites in various wildlife species. Conservation Physiology 13(1): coaf074; doi:10.1093/conphys/coaf074.2051-1434 (online)10.1093/conphys/coaf074http://hdl.handle.net/2263/107056As part of its mission to advance the field of wildlife endocrinology, the International Society of Wildlife Endocrinology aims to develop cost-effective antibodies and enzyme immunoassay kits that support research across a diverse range of species and sample matrices. To provide additional options for the quantification of faecal glucocorticoid metabolites (fGCMs), an antibody against 11-oxoetiocholanolone-17-carboxymethyl oxime (CMO) was generated in rabbits, and an enzyme immunoassay incorporating a horseradish peroxidase-conjugated label and 11-oxoetiocholanolone standard has been developed, designed for use with anti-rabbit IgG secondary antibody coated plates. This mini-kit was used to quantify glucocorticoid metabolites with a 5β-3α-ol-11-one structure in faecal extracts from 23 species: African and Asian elephants, Alpine chamois, American bison, Bengal tiger, blue wildebeest, blue-and-yellow macaw, brushtail possum, cape buffalo, fat-tailed dunnart, Florida manatee, ghost bat, giraffe, golden langur, Gould’s wattled bat, hippopotamus, Leadbeater’s possum, mandrill, okapi, roan antelope, samango monkey, short-beaked echidna, and western lowland gorilla. Pharmacological (adrenocorticotropic hormone challenge) and biological (inter-zoo translocation, wild capture, social disruption, illness/injury and veterinary intervention) challenges resulted in expected increases in fGCM concentrations, and in a subset of species, closely paralleled results from a previously established immunoassay against 11-oxoetiocholanolone-17-CMO. Two additional species tested, Krefft’s glider, which showed contradictory results on this assay compared to a previously validated enzyme immunoassay (EIA) and Ankole cow, where the magnitude increase post-event did not quite reach the 2-fold change criteria, highlight that differences in excreted faecal metabolites across species mean that no EIA will be suitable for all species. This assay provides a valuable new option for assessing adrenal activity across taxa using a group-specific antibody. Future studies should put similar emphasis on validation to determine optimal assay choice for measuring fGCMs in a variety of species.en© The Author(s) 2025. Published by Oxford University Press and the Society for Experimental Biology. This is an Open Access article distributed under the terms of the Creative Commons Attribution License.11-OxoetiocholanoloneWildlifeStressNon-invasive monitoringKit developmentFaecal glucocorticoid metabolites (fGCMs)Enzyme immunoassay (EIA)CortisolCorticosteroneArticle