**Assessment of the Stress Response in North American Deermice: Laboratory and Field Validation of Two Enzyme Immunoassays for Fecal Corticosterone Metabolites**

#### **Andreas Eleftheriou 1,\*, Rupert Palme 2 and Rudy Boonstra 3**


Received: 27 May 2020; Accepted: 26 June 2020; Published: 30 June 2020

**Simple Summary:** If we want to employ stress physiology in the managemen<sup>t</sup> and conservation of wildlife populations, we need robust methods to quantify stress physiology in the field. Although this is typically done with blood glucocorticoids (GCs), scientists now increasingly use fecal cortisol/corticosterone metabolites (FCMs), which are metabolized GCs excreted in feces. However, immunoassays to measure FCMs need to be validated for each species. North American deermice (*Peromyscus maniculatus*; hereafter deermice) are commonly used in laboratory and field studies. Although a corticosterone radioimmunoassay (RIA) has been validated for deermice, there are no validated enzyme immunoassays (EIAs), which do not require radioactive materials. Through laboratory and field experiments, we validated two EIAs for measuring FCMs in deermice. Researchers can now use these EIAs to evaluate stress physiology in deermice without the need for radioactive materials.

**Abstract:** Stress physiology is commonly employed in studies of wildlife ecology and conservation. Accordingly, we need robust and suitable methods to measure stress physiology in the field. Fecal cortisol/corticosterone metabolites (FCMs) are now increasingly being used to non-invasively evaluate adrenocortical activity; a measure of stress physiology. However, immunoassays that measure FCMs must be appropriately validated prior to their use and factors that can influence FCMs, such as trap-induced stress, must be considered. Deermice (*Peromyscus maniculatus*) are widely used in scientific studies so that developing methods that appropriately measure their adrenocortical activity is critical. In the laboratory, we tested the suitability of two enzyme immunoassays (EIAs; a corticosterone EIA, and a group-specific 5α-pregnane-3β,11β,21-triol-20-one EIA) in deermice by challenging individuals with dexamethasone and adrenocorticotropic hormone (ACTH). We found that dexamethasone suppressed FCM levels within ~10 h post injection whereas ACTH increased FCM levels within ~2 h post injection. In the field, we found that FCM levels generally increased with more time in trap confinement when using both EIAs. Although we acknowledge low sample sizes (N = 4), our results validated the two EIAs for use with FCMs from deermice.

**Keywords:** captivity-induced stress; enzyme immunoassay; fecal glucocorticoid metabolites; physiological stress in rodents
