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Faecal glucocorticoid metabolite (fGCM) analysis provides a non-invasive, feedback-free approach for monitoring adrenocortical responses to natural and anthropogenic stressors. The use of enzyme-immunoassays (EIAs) to quantify immunoreactive fGCMs has gained popularity in recent years but requires species-specific validation prior to first use. We conducted a pharmacological challenge with adrenocorticotropic hormone (ACTH) to determine whether changes in circulating glucocorticoids are reflected in fGCM, concentrations and therefore to validate excreta as a matrix for monitoring endocrine status in a southern African passerine, the White-browed Sparrow-weaver (Plocepasser mahali) and a columbid, the Laughing Dove (Spilopelia capensis). We tested the suitability of four EIAs to quantify fGCMs in 10 individuals of each species. Two of the EIAs, tetrahydrocorticosterone and 11-Oxoetiocholanolone II, detected significant elevations and were therefore most suitable for quantifying fGCMs in the White-browed Sparrow-weavers. In contrast, the 5α-pregnane-3β, 11β, 21-triol-20-one EIA detected the highest elevations in fGCM concentrations in the Laughing Doves. The lag time between stressor initiation (ACTH injection) and the resulting peak fGCM concentrations was ~2 h in both species. The validations presented here open opportunities for monitoring physiological responses in free-ranging individuals and contribute to our knowledge of the EIAs suitable for non-invasive quantification of avian fGCM concentrations.

Despite the commonly held belief that reptiles are immune to extreme temperatures, global warming is predicted to result in the loss of 40% of all reptile species by 2080. In order to understand th...

Elephant interaction programs, specifically ones that provide elephant back riding, have come under public scrutiny, and little information exists to show whether these activities affect animal wel...

Information regarding the physiology of African lions is scarce, mainly due to challenges associated with essential routine research procedures. The aim of this experiment was to test the possibility of training six captive lionesses by positive reinforcement conditioning (PRC) to voluntarily allow the collection of vaginal swabs and blood samples. This was done with the final goal of avoiding frequent anesthesia, and potentially stressful management during reproduction research. All lionesses mastered basic clicker and targeting principles within 2 weeks. Routine sampling was possible after 20 weeks of training, enabling the collection of about 750 vaginal swabs and 650 blood samples over 18 months. The animals remained calm and cooperative during all sessions, and demonstrated curiosity in the training. PRC training of captive lionesses proved to be a suitable, minimally invasive method for repeated collection of vaginal swabs and blood. Additionally, PRC may serve as behavioral enrichment for African lions in captive settings. Compared to chemical or physical restraining methods, this noninvasive management approach may reduce distress and physiological negative side effects, thus opening up new avenues for feline research.

Leopards Panthera pardus are classified in the IUCN Red List as Vulnerable, primarily due to habitat loss, natural prey base depletion and exploitation caused by various anthropogenic activities. Although protected areas are important for leopard conservation, the majority of suitable leopard habitat lies beyond protected area boundaries exposing individuals to different environmental, physiological and psychosocial stressors. This study aimed to examine the suitability of five different enzyme immunoassays (EIAs) for monitoring adrenocortical function in the leopard based on faecal glucocorticoid metabolite (fGCM) analysis. After performing an adrenocorticotrophic hormone (ACTH) stimulation test and investigating the stability of fGCM post-defaecation, faeces from free-ranging leopards in a peri-urban and a conservation area were collected to investigate the potential impact of habitat variability on glucocorticoid output. An EIA measuring fGCM with a 5α-3β-11β-diol structure performed best, demonstrating a ∼200%–330% increase in fGCM concentrations approximately 40 h post-ACTH administration. Concentrations of fGCM remained quite stable for up to six days post-defaecation and showed a maximum increase of 8% and a maximum decrease of 9%. Although not significantly different, overall median fGCM concentrations were 68% higher in individuals utilising the peri-urban area compared with leopards utilising the conservation area. The ranges of fGCM concentrations between sites, however, were similar. Individual median fGCM concentrations differed distinctly between free-ranging males and females, possibly linked to female reproductive status. The established method can now assist in addressing some of the issues facing local wildlife managers, conservationists and researchers tackling various aspects related to leopard conservation and management under different land-use practices.

Measuring physiological stress reactions through the quantification of plasma cortisol often involves physical restraint, which acts as a stressor itself. Here, we present the validation of a non-invasive method for assessing adrenocortical activity as an indicator of stress in the bat-eared fox (Otocyon megalotis). By conducting an adrenocorticotropic hormone (ACTH) challenge, we examined the suitability of three enzyme immunoassays (EIAs) detecting 11,17 dioxoandrostanes (11,17-DOA) as well as faecal glucocorticoid metabolites (fGCM) with a 5β-3α-ol- 11-one (3α,11oxo-CM), or 11,17,21-trihydroxy-4-ene-20-one structure (cortisol), respectively, for monitoring stressrelated physiological responses in male and female bat-eared foxes. Our results suggest that the cortisol EIA seems most suitable for measuring fGCMs in this myrmecophageous mammal. Using the cortisol EIA, we compared fGCM concentrations of three populations of foxes. Only one population experienced a sudden change in social environment and the assay appeared to effectively detect the expected resulting increase in stress hormone levels. Therefore, the identified EIA is effective at detecting intraspecific variation in fGCM levels and hence is a useful tool to evaluate physiological stress responses in this species.Keywords: adrenocorticotropic hormone challenge, faecal glucocorticoid metabolites, non-invasive hormone measurement, Otocyon megalotis, stress

We used vocal indicators to examine the effect of a translocation of an African elephant family herd within the Kruger National Park. These animals were moved 300 km from their home range, but returned unaided to this range within 23 days. We found that translocation resulted in a change in the mean fundamental frequency of low-frequency elephant vocalizations, known as rumbles. The rumbles increased significantly in pitch compared with pre-translocation levels during the 23 days the animals spent outside their normal home range. Mean fundamental frequency returned close to pre-translocation level by the time the animals had navigated their way back to their previous home range. Raised pitch is known to be an indicator of stress in humans and other animals. The observed acoustic results are consistent with a physiological measure of stress, faecal glucocorticoid metabolite levels, which were monitored from the same animals during the study and have already been reported elsewhere. To our knowledge, this i...

For spiny mice, detailed long-term data on reproductive endocrine function is lacking, as limited blood volumes make frequent collections of individual plasma samples for hormone monitoring impractical. To provide the necessary tools in order to fill this gap, this study examined the suitability of two enzyme immunoassays for monitoring male and female reproductive function in the southern African spiny mouse (Acomys spinosissimus) based on faecal hormone analysis. Fourteen non-pregnant and one pregnant female and 24 male spiny mice were wild-caught and subsequently monitored under controlled conditions. Thirteen of the 14 non-pregnant females revealed elevated faecal progestagen metabolite concentrations with eight individuals showing indications of a luteal phase. Two females showed two post-ovulatory luteal phases with estimated cycle lengths of 16 and 18 days, respectively. The pregnant female had an elevated (231%) mean faecal progestagen metabolite level compared with the overall mean baseline hormone concentration determined for the 14 non-pregnant females. Males exposed to a long photoperiod, simulating summer-related breeding activity, exhibited a 47.8% increase in faecal androgen metabolite levels compared with males exposed to a short photoperiod. Collectively, the data demonstrated that reproductive endocrine function can be monitored in male and female spiny mice by measuring respective faecal hormone metabolites.

This study was a preliminary investigation of an enzyme immunoassay for measuring fecal glucocorticoid metabolites in a male Asian elephant (Elephas maximus) by investigating changes in behavior and cortisol metabolite excretion associated with a putative stressful event. The study collected fecal samples for 10 days prior to, and 10 days after, 24-hr transport and relocation of the elephant to a new herd. The study measured cortisol metabolites using 2 enzyme immunoassays indicating a 389% and 340% increase in cortisol metabolite excretion following relocation. Maximal cortisol metabolite excretion occurred 2 days after relocation and remained elevated during establishment of the new herd. Stereotypic behavior increased approximately 400% after relocation. The relocation disturbed sleep patterns, the elephant spent less time sleeping during the night, and the elephant slept standing up. These results provide preliminary evidence that noninvasive monitoring of fecal cortisol metabolites can be used to investigate adrenal activity in Asian elephants and may be a safe, practical, and accurate welfare indicator.