Physiological response after translocation differs between source populations in a threatened mammal.

Conservation translocations are an important tool in the prevention of species loss, but the translocation process is associated with numerous stressors. Non-invasively monitoring stress physiology via faecal glucocorticoid metabolites (FGMs) can provide valuable insights into factors impacting translocation success and how to mitigate negative impacts. After validating an assay to measure FGMs in greater stick-nest rats (Leporillus conditor), we examined whether translocation caused a predictable change in physiology. We compared longer-term (one to five months post-translocation) physiological responses across three source populations (remnant-wild, reintroduced-wild, captive-bred), and investigated effects of body condition and sex on FGMs. Notably, FGMs of the remnant-wild population did not significantly change post-translocation, while the reintroduced-wild population exhibited a significant decrease and the captive-bred population a significant increase. Individuals in lower body condition had the highest FGMs in both wild-type populations, whereas the captive-bred population showed the opposite relationship. There was no difference in FGMs between the sexes. Our work highlights that physiological responses after translocation may not be uniform and source population history is an important factor to be considered, emphasizing the need for novel ideas that facilitate successful adaptation. By better understanding how species and individuals respond to translocation, we can improve translocation outcomes.

Sex-specific effects of a high fat diet on aortic inflammation and dysfunction.

Obesity and vascular dysfunction are independent and sexually dimorphic risk factors for cardiovascular disease. A high fat diet (HFD) is often used to model obesity in mice, but the sex-specific effects of this diet on aortic inflammation and function are unclear. Therefore, we characterized the aortic immune cell profile and function in 6-week-old male and female C57BL/6 mice fed a normal chow diet (NCD) or HFD for 10 weeks. Metabolic parameters were measured weekly and fortnightly. At end point, aortic immune cell populations and endothelial function were characterized using flow cytometry and wire myography. HFD-male mice had higher bodyweight, blood cholesterol, fasting blood glucose and plasma insulin levels than NCD mice (P < 0.05). HFD did not alter systolic blood pressure (SBP), glycated hemoglobin or blood triglycerides in either sex. HFD-females had delayed increases in bodyweight with a transient increase in fasting blood glucose at week 8 (P < 0.05). Flow cytometry revealed fewer proinflammatory aortic monocytes in females fed a HFD compared to NCD. HFD did not affect aortic leukocyte populations in males. Conversely, HFD impaired endothelium-dependent vasorelaxation, but only in males. Overall, this highlights biological sex as a key factor determining vascular disease severity in HFD-fed mice.

Scent detection dogs as a novel method for oestrus detection in an endangered species, the Tasmanian devil (Sarcophilus harrisii).

Captive breeding is a critical tool for conservation of endangered species. Identifying the correct time to pair males and females can be a major challenge for captive breeding programmes, with current methods often being invasive or slow. Detection dogs may provide a non-invasive way to determine female receptivity, but this has not been explored in captive wildlife. This exploratory study investigated the use of detection dogs as a novel method of oestrus detection in the endangered Tasmanian devil (Sarcophilus harrisii). Faecal samples were collected from 11 captive female devils during the breeding seasons of 2020 and 2021. Three dogs with prior detection experience were trained and subsequently assessed (n = 188 searches per dog), on their ability to discriminate between oestrus and non-oestrus devil faecal samples, in a one sample set-up. When assessed on training samples, dogs were able to correctly discriminate oestrus from non-oestrus with a mean sensitivity of 69.1% and mean specificity of 65.7%. When assessed on novel samples, their sensitivity to oestrus dropped (mean sensitivity of 48.6%). However, they were still able to correctly identify non-oestrus samples (mean specificity of 68.1%). This study is the first to explore detection dogs' ability to identify oestrus in a captive breeding programme for endangered wildlife, providing a promising tool for non-invasive monitoring of reproductive status in wildlife.

A light-exploiting insectivorous bat shows no melatonin disruption under lights with different spectra.

Natural light-dark cycles synchronize an animal's internal clock with environmental conditions. The introduction of artificial light into the night-time environment masks natural light cues and has the potential to disrupt this well-established biological rhythm. Nocturnal animal species, such as bats, are adapted to low light conditions and are therefore among the most vulnerable to the impacts of artificial light at night (ALAN). The behaviour and activity of insectivorous bats is disrupted by short-wavelength artificial light at night, while long-wavelength light is less disruptive. However, the physiological consequences of this lighting have not been investigated. Here, we examine the effect of LEDs with different spectra on urinary melatonin in an insectivorous bat. We collected voluntarily voided urine samples from Gould's wattled bats (Chalinolobus gouldii) and measured melatonin-sulfate under ambient night-time conditions (baseline) and under red (λP 630 nm), amber (λP 601 nm), filtered warm white (λP 586 nm) and cool white (λP 457 nm) LEDs. We found no effect of light treatment on melatonin-sulfate irrespective of spectra. Our findings suggest that short-term exposure to LEDs at night do not disrupt circadian physiology in the light-exploiting Gould's wattled bat.

The Effect of Behind-The-Scenes Encounters and Interactive Presentations on the Welfare of Captive Servals (Leptailurus serval).

The serval (Leptailurus serval) is a small African felid that is well represented in zoos and often serves as an animal ambassador in encounter programs with zoo visitors. The impact on serval welfare in relation to such programs has not been investigated to date, and the aim of this study was to assess short-term welfare effects of varying levels of visitor interaction in two captive servals. Weekly blocks of four different treatments were imposed three times on each animal over 12 weeks, and the treatments involved (1) Presentations (serval undertaking a routine training session in a designated presentation space, typically attracting high visitor numbers), (2) Behind-the-scenes (BTS, a close encounter allowing a small group of visitors to interact closely with the cat in its enclosure), (3) Presentations and BTS combined, and (4) No visitor interaction. Serval activity budgets as well as behavioural diversity were created from behaviours observed from Close Circuit Television (CCTV) footage during four daily recording sessions per animal over three consecutive days per treatment, using instantaneous scan sampling every 60 s. Individual faecal samples were collected daily to monitor changes in faecal glucocorticoid metabolite (FGM) concentration. Results indicate that the mean number of scans with stereotypic pacing was significantly reduced (p = 0.01) during Treatments 1 and 3, when cats participated in presentations only, or the two activities combined. Conversely, a significant reduction in behavioural diversity (p < 0.001) was observed when cats participated in Treatment 3, i.e., cats expressed fewer behaviours when interaction with visitors was more frequent. FGM concentrations did not vary significantly with treatment (p > 0.05). Given the reduction in stereotypic pacing, these findings suggest that involvement in an encounter program appears to exert an overall positive short-term welfare effect on the individual servals in this study. Although a reduction in behavioural diversity was not considered a negative welfare effect in the short term, potential long-term negative welfare effects resulting from a more frequent encounter program could not be ruled out in the present study.

Non-invasive monitoring of adrenocortical physiology in a threatened Australian marsupial, the western quoll (Dasyurus geoffroii).

Reintroduction has become an increasingly important conservation tool in Australia, yet the effects of stress on species during reintroduction programs have received little attention. The use of enzyme immunoassays to measure faecal glucocorticoid metabolites (FGM) is a useful non-invasive technique to monitor adrenal activity but requires validation before they can be reliably used. As part of a large reintroduction project, the goals of this study were to 1) monitor FGM in 53 western quolls (Dasyurus geoffroii) following capture from the wild and transfer to a holding facility and use this stressor to biologically validate an enzyme immunoassay; 2) determine if biological factors, such as sex, age, weight or source population affect baseline FGM levels; and 3) examine individual variation in the acute adrenal response of quolls to the capture and transfer associated with reintroductions. We successfully validated an assay that targets glucocorticoid metabolites with a 5α-3ß,11ß-diol structure and found that sex significantly influenced both baseline and peak FGM output in western quolls, whereas age, weight and source population did not. We also observed considerable variation among individuals in the magnitude and duration of their physiological response to capture and transfer. Using the methods described here, FGM analysis may provide further information about the adrenal activity of the western quoll and improve future conservation efforts for this threatened species.

Effects of Regulating Visitor Viewing Proximity and the Intensity of Visitor Behaviour on Little Penguin (Eudyptula minor) Behaviour and Welfare.

We examined the effects of regulating the viewing proximity of visitors and the intensity of visitor behaviour on behaviours indicative of fear and stress physiology of 15 zoo-housed little penguins (Eudyptula minor). A 2 × 2 factorial fully randomised design was used to examine the effects of regulating: 1) the viewing proximity of visitors to enclosure, 'normal viewing distance' and 'increased viewing distance' (using a physical barrier set up 2 m away from the enclosure), and 2) the intensity of visitor behaviour, 'unregulated visitor behaviour' and 'regulated visitor behaviour' (using signage and uniformed personnel). In addition, a treatment consisting of closing the enclosure to visitors was included. Penguin behaviour, visitor numbers and visitor behaviour were recorded by CCTV video recordings and direct observations, respectively. Penguin faecal glucocorticoid metabolites were also analysed as a measure of stress physiology. We found that increased viewing distance reduced (p < 0.05) all visitor behaviours except for loud vocalisations and tactile contact with penguins. However, there were no direct effects of signage and uniformed personnel on visitor behaviour (p > 0.05). As the regulation of viewing proximity increased from a closed exhibit to an open exhibit with increased viewing distance, and then to an open exhibit with normal viewing distance, this increased the proportion of penguins huddling (p = 0.0011), vigilant (p = 0.0060) and retreating (p = 0.00013), and decreased the proportion of penguins within 1 m of the visitor viewing area (p = 0.00066), surface swimming (p = 0.00091) and preening in the water (p = 0.042). There were also limited effects of regulating visitor behaviour on penguin behaviour. No treatment effects were found on faecal glucocorticoid metabolites (p > 0.05). These results indicate that regulating visitor viewing proximity affects penguin behaviours indicative of fear and visitor behaviour. This suggests that close visitor contact can be fear-provoking for little penguins but increasing the distance between visitors and penguins can reduce fear responses of penguins by regulating both viewing proximity and visitor behaviour. However, it is unclear whether these changes in penguin behaviour are due to the increased separation between visitors and penguins and/or specific visitor behaviours associated with close viewing proximity to the enclosure, such as leaning over the enclosure or tactile contact with the pool, which are impeded when visitors are further away.

Meta-analytic insights into factors influencing the repeatability of hormone levels in agricultural, ecological, and medical fields.

Interest in individual variation in hormone concentrations is rapidly increasing, particularly with regard to the evolutionary and practical implications. A key aspect of studying individual variation in any labile trait is estimating the degree of within- versus among-individual variation, but at present, we do not have a broad consensus on the extent to which hormone levels are repeatable and what factors might influence repeatability. To address this knowledge gap, we conducted a comprehensive meta-analysis of hormone levels that included 1,132 estimates of repeatability from 368 studies across three fields of study: agriculture, ecology, and medicine. We assessed the influence of sex, age class, sample type, hormone family, type of hormone measure, assay type, number of subjects, number of samples per subject, and sampling interval on repeatability estimates. Overall mean repeatability was 0.58, but estimates differed substantially among study disciplines, being lowest in ecology (0.34), moderate in agriculture (0.52), and relatively high in medicine (0.68). In addition, repeatability decreased slightly as sampling interval increased, and also tended to be higher for peak hormone levels than baseline levels. Overall, hormone levels are moderately repeatable, suggesting that they can potentially be useful indicators of individual variation. However, estimates of repeatability are quite variable among fields, so caution should be used when relying on single samples to assess individual variation.

Effects of invasion history on physiological responses to immune system activation in invasive Australian cane toads.

The cane toad (Rhinella marina) has undergone rapid evolution during its invasion of tropical Australia. Toads from invasion front populations (in Western Australia) have been reported to exhibit a stronger baseline phagocytic immune response than do conspecifics from range core populations (in Queensland). To explore this difference, we injected wild-caught toads from both areas with the experimental antigen lipopolysaccharide (LPS, to mimic bacterial infection) and measured whole-blood phagocytosis. Because the hypothalamic-pituitary-adrenal axis is stimulated by infection (and may influence immune responses), we measured glucocorticoid response through urinary corticosterone levels. Relative to injection of a control (phosphate-buffered saline), LPS injection increased both phagocytosis and the proportion of neutrophils in the blood. However, responses were similar in toads from both populations. This null result may reflect the ubiquity of bacterial risks across the toad's invaded range; utilization of this immune pathway may not have altered during the process of invasion. LPS injection also induced a reduction in urinary corticosterone levels, perhaps as a result of chronic stress.

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.

Stress-induced peak (but not resting) metabolism correlates with mating display intensity in male guppies.

Recent empirical and conceptual papers have highlighted the potential for metabolism to act as a proximate mechanism for behavior that could explain animal personality (consistency over time). Under this hypothesis, individuals with consistently high levels of behavioral activity should also have high resting metabolic rate (RMR) as it can reflect capacity to process food and generate energy. We tested for the predicted positive covariance between RMR and three behaviors that differ in energy demands in 30 male guppies, using multivariate mixed models; we repeatedly measured their activity (10 times each), courtship displays (nine times), voracity (10 times), and metabolism (four-times). Resting metabolic rate (measured overnight in respirometry trials) did not consistently differ among males, whereas initial peak metabolism measured during those same trials (R = 0.42), and all behaviors were repeatable (R = 0.33-0.51). RMR declined over time suggesting habituation to the protocol, whereas peak metabolism did not. Initial peak metabolism was negatively correlated with courtship display intensity, and voracity was positively correlated with activity, but all other among-individual correlations were not significant. We conclude that RMR does not provide a proximate explanation for consistent individual differences in behavior in male guppies, and therefore the potential for independent evolution of these physiological and behavioral traits seems possible. Finally, we identify peak metabolism as a potential measure of the stress response to confinement, which highlights the value of considering various aspects of metabolic rates recording during respirometry trials.

The value of eutherian-marsupial comparisons for understanding the function of glucocorticoids in female mammal reproduction.

This article is part of a Special Issue "SBN 2014". Chronic stress is known to inhibit female reproductive function. Consequently, it is often assumed that glucocorticoid (GC) concentrations should be negatively correlated with reproductive success because of the role they play in stress physiology. In contrast, a growing body of evidence indicates that GCs play an active role in promoting reproductive function. It is precisely because GCs are so integral to the entire process that disruptions to adrenal activity have negative consequences for reproduction. The goal of this paper is to draw attention to the increasing evidence showing that increases in adrenal activity are important for healthy female reproduction. Furthermore, we outline several hypotheses about the functional role(s) that GCs may play in mediating reproduction and argue that comparative studies between eutherian and marsupial mammals, which exhibit some pronounced differences in reproductive physiology, may be particularly useful for testing different hypotheses about the functional role of GCs in reproduction. Much of our current thinking about GCs and reproduction comes from research involving stress-induced levels of GCs and has led to broad assumptions about the effects of GCs on reproduction. Unfortunately, this has left a gaping hole in our knowledge about basal GC levels and how they may influence reproductive function, thereby preventing a broader understanding of adrenal physiology and obscuring potential solutions for reproductive dysfunction.

Impacts of visitor number on Kangaroos housed in free-range exhibits.

Free range exhibits are becoming increasingly popular in zoos as a means to enhance interaction between visitors and animals. However very little research exists on the impacts of visitors on animal behaviour and stress in free range exhibits. We investigated the effects of visitor number on the behaviour and stress physiology of Kangaroo Island (KI) Kangaroos, Macropus fuliginosus fuliginosus, and Red Kangaroos, Macropus rufus, housed in two free range exhibits in Australian zoos. Behavioural observations were conducted on individual kangaroos at each site using instantaneous scan sampling to record activity (e.g., vigilance, foraging, resting) and distance from the visitor pathway. Individually identifiable faecal samples were collected at the end of each study day and analysed for faecal glucocorticoid metabolite (FGM) concentration. When visitor number increased, both KI Kangaroos and Red Kangaroos increased the time spent engaged in visitor-directed vigilance and KI Kangaroos also increased the time spent engaged in locomotion and decreased the time spent resting. There was no effect of visitor number on the distance kangaroos positioned themselves from the visitor pathway or FGM concentration in either species. While there are limitations in interpreting these results in terms of fear of visitors, there was no evidence of adverse effects animal welfare in these study groups based on avoidance behaviour or stress physiology under the range of visitor numbers that we studied.

Gender differences in conference presentations: a consequence of self-selection?

Women continue to be under-represented in the sciences, with their representation declining at each progressive academic level. These differences persist despite long-running policies to ameliorate gender inequity. We compared gender differences in exposure and visibility at an evolutionary biology conference for attendees at two different academic levels: student and post-PhD academic. Despite there being almost exactly a 1:1 ratio of women and men attending the conference, we found that when considering only those who presented talks, women spoke for far less time than men of an equivalent academic level: on average student women presented for 23% less time than student men, and academic women presented for 17% less time than academic men. We conducted more detailed analyses to tease apart whether this gender difference was caused by decisions made by the attendees or through bias in evaluation of the abstracts. At both academic levels, women and men were equally likely to request a presentation. However, women were more likely than men to prefer a short talk, regardless of academic level. We discuss potential underlying reasons for this gender bias, and provide recommendations to avoid similar gender biases at future conferences.

Cyclic changes in cortisol across the estrous cycle in parous and nulliparous Asian elephants.

In the context of reproduction, glucocorticoids (GCs) are generally considered to have negative effects. However, in well-studied model species, GCs fluctuate predictability across the estrous cycles, and short-term increases promote healthy ovarian function. Reproductive challenges have plagued captive elephant populations, which are not currently self-sustaining. Efforts to understand reproductive dysfunction in elephants have focused on the suppressive effects of cortisol, but the potential permissive or stimulatory effects of cortisol are unknown. In this study, we provide a detailed examination of cortisol patterns across the estrous cycle in Asian elephants (Elephas maximus). Time series analysis was used to analyze cortisol and progesterone data for a total of 73 cycles from eight females. We also compared cortisol profiles between females that successfully conceived and females that failed to conceive despite repeated mating attempts. Our results revealed that cortisol fluctuates predictably across the estrous cycle, with a peak during the second half of the follicular phase followed by low levels throughout the luteal phase. Furthermore, this pattern was significantly altered in nulliparous females; cortisol concentrations did not decline during the luteal phase to the same extent as in parous females. This study highlights the complexity of cortisol signaling and suggests future directions for understanding the role of cortisol in reproductive dysfunction.

Cost of reproduction in the Queensland fruit fly: Y-model versus lethal protein hypothesis.

The trade-off between lifespan and reproduction is commonly explained by differential allocation of limited resources. Recent research has shown that the ratio of protein to carbohydrate (P : C) of a fly's diet mediates the lifespan-reproduction trade-off, with higher P : C diets increasing egg production but decreasing lifespan. To test whether this P : C effect is because of changing allocation strategies (Y-model hypothesis) or detrimental effects of protein ingestion on lifespan (lethal protein hypothesis), we measured lifespan and egg production in Queensland fruit flies varying in reproductive status (mated, virgin and sterilized females, virgin males) that were fed one of 18 diets varying in protein and carbohydrate amounts. The Y-model predicts that for sterilized females and for males, which require little protein for reproduction, there will be no effect of P : C ratio on lifespan; the lethal protein hypothesis predicts that the effect of P : C ratio should be similar in all groups. In support of the lethal protein hypothesis, and counter to the Y-model, the P : C ratio of the ingested diets had similar effects for all groups. We conclude that the trade-off between lifespan and reproduction is mediated by the detrimental side-effects of protein ingestion on lifespan.

Comparative patterns of adrenal activity in captive and wild Canada lynx (Lynx canadensis).

Stress and animal well-being are often assessed using concentrations of glucocorticoids (GCs), a product of the hypothalamic-pituitary-adrenal axis. However, GC concentrations can also be modulated by predictable events, such as changes in season or life history stage. Understanding normative patterns of adrenal activity is critical for making valid conclusions about changes in GC concentrations. In this study, we validated an assay for monitoring fecal glucocorticoid metabolites (FGM) in Canada lynx. We then used this technique to assess patterns of adrenal activity in Canada lynx across several contexts. Our results show that captive lynx have higher FGM concentrations than wild lynx, which may be related to differences in stress levels, metabolic rate, diet, or body condition. We also found that FGM concentrations are correlated with reproductive status in females, but not in males. For males, seasonal increases in FGM expression coincide with the onset of the breeding season, whereas in females, FGM increase toward the end of the breeding season. This information provides a valuable foundation for making inferences about normative versus stress-induced changes in adrenal activity in Canada lynx.

Patterns of testicular activity in captive and wild Canada lynx (Lynx canadensis).

Canada lynx are listed as a threatened species in the contiguous US. Understanding the reproductive characteristics (i.e., mating system, behavior, physiology) of a species is useful for ensuring effective in situ and ex situ management plans. The goal of this study was to describe patterns of androgen expression in both captive and wild male Canada lynx using fecal hormone metabolite analysis. Among captive lynx, juvenile and castrated males had lower concentrations of fecal androgens (fA) than intact males, thereby demonstrating that the assay detects biologically meaningful differences in testicular activity. We found that captive males in general had much higher fA levels than wild males. All males showed strong seasonal variation in fA concentrations, with significantly higher levels being expressed during the breeding season (February and March) than during the non-breeding season. Among captive males, variation in seasonal fA levels did not correlate with latitude. Finally, males housed with intact cage-mates (either male or female) had significantly higher fA levels than males housed alone or with a neutered cage-mate.

Patterns of ovarian and luteal activity in captive and wild Canada lynx (Lynx canadensis).

Canada lynx face some unique breeding restrictions, which may have implications for population viability and captive management. The goal of this study was to improve our understanding of basic reproductive physiology in Canada lynx. Using fecal hormone metabolite analysis, we established normative patterns of fecal estrogen (fE) and progestagen (fP) expression in captive and wild female Canada lynx. Our results indicate that Canada lynx have persistent corpora lutea, which underlie their uncharacteristic fP profiles compared to other felids. Thus, fP are not useful for diagnosing pregnancy in Canada lynx. We also found that Canada lynx are capable of ovulating spontaneously. Captive females had higher concentrations of fE and fP than wild females. Both populations exhibit a seasonal increase in ovarian activity (as measured by fE) between February and April. Finally, there was evidence of ovarian suppression when females were housed together.