Validation of an Enzyme Immunoassay to Measure Faecal Glucocorticoid Metabolites in Common Brushtail Possums (Trichosurus vulpecula) to Evaluate Responses to Rehabilitation

SIMPLE SUMMARY: Little is known about how exposure to novel stimuli during rescue and rehabilitation could affect the physiology of native wildlife. We investigated this question in a species commonly rescued for rehabilitation, the common brushtail possum (Trichosurus vulpecula). Glucocorticoids (major hormones involved in stress responses) are metabolised in the body and excreted in the form of faecal glucocorticoid metabolites, which can be measured as a way of evaluating the response of animals to potential stressors. Comparing five enzyme immunoassay options, we found that the 11-oxoaetiocholanolone (abbreviation: 72a) EIA was the most suitable for measuring these metabolites in brushtail possums. This assay was then used to measure faecal glucocorticoid metabolite concentrations in 20 possums during rehabilitation. The probability of a physiological “stress” response occurring within five days of a potentially stressful event was about 50%, regardless of the type of event. There was a high level of variation in hormone profiles between possums. Our study has demonstrated that injured and orphaned possums show detectable changes in faecal glucocorticoid metabolites during captivity and rehabilitation, and has identified events that can induce a physiological response in some individuals. This is the first step toward understanding the relationship between these responses during rehabilitation and survival. ABSTRACT: Volunteer wildlife rehabilitators rescue and rehabilitate thousands of native animals every year in Australia. However, there is little known about how exposure to novel stimuli during rehabilitation could affect the physiology of wildlife. We investigated this question in a species that commonly enters rehabilitation, the common brushtail possum (Trichosurus vulpecula). We evaluated five enzyme immunoassays (EIA) to determine the most suitable for measuring faecal glucocorticoid metabolites (FGM) as a proxy for evaluating the response of brushtail possums to potential stressors during rehabilitation. An adrenocorticotrophin hormone (ACTH) challenge was conducted on wild-caught possums to determine the best-performing EIA based on the successful detection of FGM peaks in at least two of three possums. While a number of assays met these criteria, the 11-oxoaetiocholanolone (abbreviation: 72a) EIA was selected as it had the largest amplitude of change in response to the ACTH challenge. This assay was then used to measure FGM concentrations in 20 possums during rehabilitation. There was high variation in baseline FGM concentrations and response to captivity between possums. Significant changes in FGM levels were detected in most possums during captivity, but were not reliably associated with potentially stressful events that were identified by rehabilitators. The probability of an FGM peak occurring within five days of a potentially stressful event was about 50%, regardless of the type of event. Our study has demonstrated that injured and orphaned possums show changes in FGMs during captivity and rehabilitation and has identified events that can induce a physiological response in some individuals. We recommend that research now focus on the relationship between these responses during rehabilitation and pre- and post-release survival.