Using Faecal Cortisol Metabolites to Assess Adrenocortical Activity in Wild‐Living Alpine Marmot <scp> <i>Marmota marmota</i> </scp> : A Biological Validation Experiment

Faecal cortisol/corticosterone metabolites (FCMs) have become increasingly popular as an easy-to-sample, non-invasive and feedback-free alternative to assess glucocorticoid (GC) levels, key components of the neuroendocrine stress response and other physiological processes. While FCMs can be a powerful aid, for instance, for gaining insights into ecological and evolutionary processes, as well as to assess animal welfare or impacts of anthropogenic stressors on wildlife populations, this method comes with specific challenges. Because GCs are heavily metabolised before excretion, it is critical to validate the enzyme immunoassays (EIAs) used to measure FCMs. Additionally, because species may differ in metabolite profiles, assay validation must be performed separately for each focal species. Despite this, the use of unvalidated assays remains widespread. We performed a biological validation experiment to test a set of EIAs to measure FCMs and adrenocortical activity in free-living Alpine marmots <i>Marmot marmota</i>. We capitalised on capture and handling as part of a relocation project of marmots under the assumption that capture, and handling represent a stressful event and tracked changes in FCM levels over the following 48 h. Faeces samples collected at capture were assumed to return baseline FCM levels. Of the three EIAs tested, only the 11-oxoetiocholanolone '72T' EIA detected an increase in FCM levels about 18 h after capture. This result paves the way for future studies using FCMs to investigate the adrenocortical activity in this species.