Successful acclimatization of mandrills (Mandrillus sphinx) translocated to Conkouati-Douli National Park, Republic of Congo, as measured by fecal glucocorticoid metabolites

Translocation and reintroduction are common tools in conservation management and can be very successful. However, translocation can be stressful for the animals involved, and stress is implicated as a major cause of failure in release programs. Conservation managers should therefore seek to understand how the stages of translocation impact stress physiology in the animals involved. We quantified fecal glucocorticoid metabolites (fGCMs) as a noninvasive measure of response to potential stressors during a translocation of 15 mandrills (Mandrillus sphinx) into Conkouati-Douli National Park, Republic of Congo. The mandrills were initially housed in a sanctuary, transferred to a pre-release enclosure in the National Park and then released into the forest. We collected repeated fecal samples (n = 1101) from known individuals and quantified fGCMs using a previously validated enzyme immunoassay. Transfer from the sanctuary to the pre-release enclosure correlated with a significant 1.93-fold increase in fGCMs, suggesting that transfer was a stressor for the mandrills. fGCM values decreased over time in the pre-release enclosure, suggesting that the mandrills recovered from the transfer and acclimatized to the enclosure. Release to the forest was not linked to a significant increase in fGCMs over the final values in the enclosure. Following release, fGCMs continued to decrease, fell below sanctuary values after just over a month and were about half the sanctuary values after 1 year. Overall, our results suggest that the translocation, although initially presenting a physiological challenge to the animals, was not detrimental to the well-being of the animals over the timescale of the study and, in fact, may have been beneficial. Our findings show the value of non-invasive physiology in monitoring, evaluating and designing wildlife translocations and, ultimately, contributing to their success

Simulated heatwave and fishing stressors alter corticosteroid and energy balance in neonate blacktip reef sharks, Carcharhinus melanopterus

The increasing frequency and duration of marine heatwaves attributed to climate change threatens coastal elasmobranchs and may exacerbate existing anthropogenic stressors. While the elasmobranch stress response has been well studied, the role of the unique corticosteroid-1α-hydroxycorticosterone (1α-OHB)-in energy balance is not understood. Therefore, 1α-OHB's utility as a stress biomarker in elasmobranch conservation physiology is equivocal. Here, we analyse the roles of corticosteroids, 1α-OHB and corticosterone, and metabolites, glucose and 3-hydroxybutyrate (3-HB), in response to stress in a protected tropical shark species, the blacktip reef shark (Carcharhinus melanopterus). Wild-caught neonates were exposed to ambient (27°C) or heatwave conditions (29°C) and subsequently a simulated fishing stressor (1 min air exposure). Blood samples were taken prior to temperature exposure, prior to air exposure, and 30 min, 1 h, 24 h, and 48 h post-Air exposure at treatment temperatures. Plasma 1α-OHB was elevated for 48 h in 27°C-exposed sharks but declined over time in 29°C-exposed sharks. Plasma 1α-OHB was not correlated with either metabolite. Plasma glucose was higher and plasma 3-HB was lower in 29°C-exposed sharks. In a separate experiment, blood samples were collected from both neonate and adult sharks immediately following capture and again 5 min later, and analysed for corticosteroids and metabolites. Plasma 1α-OHB increased in neonates within 5 min, but neonates displayed lower plasma 1α-OHB and higher glucose concentrations than adults. We conclude that 1α-OHB does not serve as a classic glucocorticoid role in C. melanopterus under these stressors. Furthermore, we show for the first time, ontogenetic differences in plasma 1α-OHB. Ultimately, our findings provide insights into hormonal control of energy mobilization during stress in C. melanopterus, particularly during simulated heatwave conditions, which seem to alter both endocrine and energy mobilization. Further work is needed to determine the utility of 1α-OHB as a biomarker for the mobilization of energy during a stress event in elasmobranchs

Biochemical and biological validations of a faecal glucocorticoid metabolite assay in mandrills (Mandrillus sphinx)

Stress is a major factor in determining success when releasing endangered species into the wild but is often overlooked. Mandrills (Mandrills sphinx) are vulnerable to extinction due to habitat loss and demand for bush meat and the pet trade. To help bolster in situ populations, rehabilitated rescued mandrills recently were released into a protected area in the Republic of Congo. The goal of this study was to validate the use of faecal glucocorticoid metabolite enzyme immunoassays (EIAs) in mandrills and test field-friendly faecal hormone extraction techniques that can subsequently be used to monitor the stress physiology and welfare of mandrills throughout the release process. Using faecal samples collected from ex situ mandrills, we tested cortisol, corticosterone, 11β-hydroxyetiocholanolone (69a), and 11-oxoetiocholanolone EIAs. Absolute concentrations, hormone profiles following medical procedures or translocation, and high-performance liquid chromatography fraction immunoreactivity showed that the 69a assay was the best choice to monitor the stress response in this species. Samples with delayed extraction or drying times had 40–80% lower 69a concentrations than samples extracted immediately post-collection and frozen. The 69a EIA is an appropriate assay for monitoring welfare in this species in situ or ex situ, and results indicated that consistent extraction methods are important for accurate comparisons

Challenges, Pitfalls and Surprises: Development and Validation of a Monoclonal Antibody for Enzyme Immunoassay of the Steroid 1α-Hydroxycorticosterone In Elasmobranch Species

Sharks and rays are popular species used in wildlife ecotourism and aquariums to educate the public on the behavior, ecology and conservation challenges of elasmobranchs. To understand long-term physiological health and welfare under varying social and husbandry conditions, we developed and validated an enzyme immunoassay (EIA) to measure stress/ionoregulatory hormones in managed and semi-free range southern rays (Hypanus americanus). Banked serum and interrenal samples from 27 female rays managed at Disney’s The Seas with Nemo and Friends® and Castaway Cay were used to evaluate measurement of 1α-hydroxycorticosterone (1αOHB) relative to corticosterone (B). Although commercial EIAs are available for B, those tested exhibit only low relative cross-reactivity to 1αOHB (3-5%). To improve measurement of 1αOHB, we developed a monoclonal antibody using a synthesized 1αOHB-derivative for evaluation using high-performance liquid chromatography (HPLC) and EIA. Relative displacements of cross-reactant compounds showed that the antibody had good sensitivity for the target antigen 1αOHB, and low sensitivity to related steroids (desoxycorticosterone and B), but greater sensitivity to 11-dehydrocorticosterone. Tests of competitive vs. noncompetitive EIA formats, reagent titration, and incubation times of the antibody and conjugate were used to optimize sensitivity, repeatability and precision of measured 1αOHB in standards and samples (4 ng/ml, 90% binding). Tests of sample pre-treatment (pH adjustment) and extraction with varying solvent polarity were used to optimize measurement of 1αOHB i