title: Non-invasive monitoring of physiological stress in the Western lowland gorilla (Gorilla gorilla gorilla): validation of a fecal glucocorticoid assay and methods for practical application in the field.
abstract: Enzymeimmunoassays (EIAs) allow researchers to monitor stress hormone output via measurement of fecal glucocorticoid metabolites (FGCMs) in many vertebrates. They can be powerful tools which allow the acquisition of otherwise unobtainable physiological information from both captive animals and wild animals in remote forest habitats, such as great apes. However, methods for hormone measurement, extraction and preservation need to be adapted and validated for field settings. In preparation for a field study of Western lowland gorillas (Gorilla gorilla gorilla) in the Central African Republic we used samples from captive gorillas collected around opportunistic stressful situations to test whether four different glucocorticoid EIAs reflected adrenocortical activity reliably and to establish the lag-time from the stressor to peak excretion. We also validated a field extraction technique and established a simple, non-freezer-reliant method to preserve FGCMs in extracts long-term. We determined the rate of FGCM change over 28 days when samples cannot be extracted immediately and over 12h when feces cannot be preserved immediately in alcohol. Finally, we used repeat samples from identified individuals to test for diurnal variation in FGCM output. Two group-specific assays measuring major cortisol metabolites detected the predicted FGCM response to the stressor reliably, whereas more specific cortisol and corticosterone assays were distinctly less responsive and thus less useful. We detected a lag time of 2-3 days from stressor to peak FGCM excretion. Our field extraction method performed as well as an established laboratory extraction method and FGCMs in dried extracts stored at ambient temperatures were as stable as those at -20 °C over 1 yr. Hormones in non-extracted feces in alcohol were stable up to 28 days at ambient temperatures. FGCMs in un-fixed gorilla feces deteriorated to almost 50% of the original values within 6h under field conditions. We detected no diurnal variation in FGCMs in samples from wild gorillas. Our study highlights the importance of thorough biological and immunological validation of FGCM assays, and presents validated, practical methods for the application of non-invasive adrenocortical monitoring techniques to field conservation contexts where it is crucially needed.
authors.affiliation: Department of Anthropology, Durham University, DH1 3LE, Durham, UK. k.a.shutt@durham.ac.uk
authors.forename: Kathryn
authors.initials: K
authors.lastname: Shutt
authors.forename: Joanna M
authors.initials: JM
authors.lastname: Setchell
authors.forename: Michael
authors.initials: M
authors.lastname: Heistermann
mesh_terms: D000818:Animals; D000835:Animals, Wild / Q000502:physiology; D000836:Animals, Zoo / Q000502:physiology*; D002488:Central African Republic; D002940:Circadian Rhythm; D000431:Ethanol; D005243:Feces / Q000737:chemistry*; D005260:Female; D005938:Glucocorticoids / Q000032:analysis*; D006071:Gorilla gorilla / Q000502:physiology*; D007124:Immunoenzyme Techniques; D008297:Male; D013048:Specimen Handling / Q000379:methods; D013312:Stress, Physiological
chemical_list: D005938:Glucocorticoids; D000431:Ethanol