Abstract:As 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.