One size does not fit all: Monitoring faecal glucocorticoid metabolites in marsupials.

Marsupial research, conservation, and management can benefit greatly from knowledge about glucocorticoid (GC) secretion patterns because GCs influence numerous aspects of physiology and play a crucial role in regulating an animal's response to stressors. Faecal glucocorticoid metabolites (FGM) offer a non-invasive tool for tracking changes in GCs over time. To date, there are relatively few validated assays for marsupials compared with other taxa, and those that have been published generally test only one assay. However, different assays can yield very different signals of adrenal activity. The goal of this study was to compare the performance of five different enzyme immunoassays (EIAs) for monitoring adrenocortical activity via FGM in 13 marsupial species. We monitored FGM response to two types of events: biological stressors (e.g., transport, novel environment) and pharmacological stimulation (ACTH injection). For each individual animal and assay, FGM peaks were identified using the iterative baseline approach. Performance of the EIAs for each species was evaluated by determining (1) the percent of individuals with a detectable peak 0.125-4.5days post-event, and (2) the biological sensitivity of the assay as measured by strength of the post-event response relative to baseline variability (Z-score). Assays were defined as successful if they detected a peak in at least 50% of the individuals and the mean species response had a Z⩾2. By this criterion, at least one assay was successful in 10 of the 13 species, but the best-performing assay varied among species, even those species that were closely related. Furthermore, the ability to confidently assess assay performance was influenced by the experimental protocols used. We discuss the implications of our findings for biological validation studies.

Wombat reproduction (Marsupialia; Vombatidae): an update and future directions for the development of artificial breeding technology.

This review provides an update on what is currently known about wombat reproductive biology and reports on attempts made to manipulate and/or enhance wombat reproduction as part of the development of artificial reproductive technology (ART) in this taxon. Over the last decade, the logistical difficulties associated with monitoring a nocturnal and semi-fossorial species have largely been overcome, enabling new features of wombat physiology and behaviour to be elucidated. Despite this progress, captive propagation rates are still poor and there are areas of wombat reproductive biology that still require attention, e.g. further characterisation of the oestrous cycle and oestrus. Numerous advances in the use of ART have also been recently developed in the Vombatidae but despite this research, practical methods of manipulating wombat reproduction for the purposes of obtaining research material or for artificial breeding are not yet available. Improvement of the propagation, genetic diversity and management of wombat populations requires a thorough understanding of Vombatidae reproduction. While semen collection and cryopreservation in wombats is fairly straightforward there is currently an inability to detect, induce or synchronise oestrus/ovulation and this is an impeding progress in the development of artificial insemination in this taxon.

Eliminative behavior of captive numbats, Myrmecobius fasciatus (Mammalia; Marsupialia): pattern and identification of fecal deposits.

This study examined the spatial defecation patterns of numbats within captivity and tested the efficacy of a food colorant as a fecal marker in this species. Rather than randomly distributing scats throughout their environment, the numbats aggregated their feces at specific sites forming latrines. It is unclear whether the strong inclination for latrine formation was due to this species' inherent behavior or is a direct result of captivity. Males were found to have a higher daily defecation rate, different defecation time, larger number of latrines, and greater number of scats per quadrant, as compared to females. In this study, the majority of scats were deposited along enclosure boundaries and for both sexes there was a higher probability of latrines being placed along enclosure fencing shared with a female neighbor. The high concentration of latrines along boundaries suggests that they may play a role in territorial defense. The results also indicated that captive numbats tend to choose defecation points away from food and refuge sites. Transit and total retention time of the marker through the GIT was ≤3 hr and between 24 and 27 hr, respectively. A marker dose of 3 gtts feed(-1) × 2 feeds day(-1) was required to reach a steady and detectable state of marker output, which enabled accurate identification of individual samples during the breeding season. Reliable labeling was obtained using blue and red colored food dye, and there was no evidence that incorporation of the marker into the diet had any negative effects on food intake.

Non-invasive assessment of stress in captive numbats, Myrmecobius fasciatus (Mammalia: Marsupialia), using faecal cortisol measurement.

Annual patterns of faecal cortisol metabolite (FCM) secretion were examined in six captive numbats (Myrmecobius fasciatus). The use of enzyme-immunoassay for the measurement of FCM in the numbat faeces was validated using an adrenocorticotropic hormone (ACTH) challenge and the resultant FCM measurements represent the first description of adrenal endocrinology in this species. Total overall, baseline and peak FCM mean concentrations varied according to individual, but not gender. For males, mean baseline and overall FCM secretion was higher in spring in summer (compared to winter and autumn) and was elevated during the breeding season. For females, mean baseline FCM secretion did not differ by season or breeding season, but mean overall FCM secretion was elevated during the breeding season. Thus, male (but not female) numbats display an annual change in FCM secretion that is strongly linked to their seasonal pattern of reproduction. Significant FCM elevations (n=178) were observed in response to 20 different stressors, with these stressors being allocated to one of six categories: ANIM, ENVIRO, HAND, HEALTH, MAN and UNK. The mean proportion of positive responses to each category varied according to category, season and breeding season, but did not vary by individual or gender. ANIM and HEALTH stressors elicited a higher response rate than all other categories and an increase in the number of ANIM, ENVIRO, and HEALTH stressors were observed during the breeding season. Although there were multiple stressors within the captive environment that the numbats reacted to, this did not translate into a welfare issue.