Search

Funder: University of Zurich; Id: http://dx.doi.org/10.13039/501100006447, Funder: MAVA Foundation, Funder: Mammal Research Institute at the University of Pretoria, The ability of dispersing individuals to adjust their behaviour to changing conditions is instrumental in overcoming challenges and reducing dispersal costs, consequently increasing overall dispersal success. Understanding how dispersers' behaviour and physiology change during the dispersal process, and how they differ from resident individuals, can shed light on the mechanisms by which dispersers increase survival and maximise reproduction. By analysing individual behaviour and concentrations of faecal glucocorticoid metabolites (fGCM), a stress-associated biomarker, we sought to identify the proximate causes behind differences in survival and reproduction between dispersing and resident meerkats Suricata suricatta. We used data collected on 67 dispersing and 108 resident females to investigate (a) which individual, social and environmental factors are correlated to foraging and vigilance, and whether the role of such factors differs among dispersal phases, and between dispersers and residents; (b) how time allocated to either foraging or vigilance correlated to survival in dispersers and residents and (c) the link between aggression and change in fGCM concentration, and their relationship with reproductive rates in dispersing groups and resident groups with either long-established or newly established dominant females. Time allocated to foraging increased across dispersal phases, whereas time allocated to vigilance decreased. Time allocated to foraging and vigilance correlated positively and negatively, respectively, with dispersers' group size. We did not find a group size effect for residents. High proportions of time allocated to foraging correlated with high survival, and more so in dispersers, suggesting that maintaining good physical condition may reduce mortality during dispersal. Furthermore, while subordinate individuals rarely reproduced in resident groups, the conception rate of subordinates in newly formed dispersing groups was equal to that of their dominant individuals. Mirroring conception rates, in resident groups, fGCM concentrations were lower in subordinates than in dominants, whereas in disperser groups, fGCM concentrations did not differ between subordinates and dominants. Our results, which highlight the relationship between behavioural and physiological factors and demographic rates, provide insights into some of the mechanisms that individuals of a cooperative species can use to increase overall dispersal success.

Quantification of glucocorticoids and their metabolites as a central component of the physiological stress response has many potential applications in ornithological research, but require careful species-specific validations. We established a reliable non-invasive method to measure faecal glucocorticoid metabolites (fGCM) in the southern yellow-billed hornbill (Tockus leucomelas). Following an adrenocorticotropic hormone (ACTH) challenge, plasma corticosterone levels increased by 110%, with a tetrahydrocorticosterone enzyme immunoassay (EIA) revealing a ~ fourfold increase in fGCM concentrations approximately 3 h post-injection. These data reveal droppings are a suitable matrix for monitoring stress-associated biomarkers in this species and suggest the tetrahydrocorticosterone EIA may often be appropriate for wild birds.

Little information exists on the sexual behaviours of the roan antelope (Hippotragus equinus; E. Geoffroy Saint-Hilaire, 1803), a species listed vulnerable in southern Africa. To fill this gap, two adult males and 13 adult females from two captive breeding herds, at Lapalala Wilderness, South Africa, were observed three times per day from September 2017 to July 2018. Pre-categorised behaviours exhibited by males and females were recorded and their frequencies quantified. Fifteen pre-defined behaviours were recorded in males with precopulatory behaviours being the most recurrent. Overall, 181 courtships and 73 complete mounts (copulations) were recorded for the two mating periods observed; each mount lasted between 3 and 4 s. In females, eight pre-defined behaviours were observed and homosexual mounts were observed for the first time in females of this species and indeed within the Hippotragini tribe. Receptivity lasted for between 24 and 72 h, with prominent behaviours being copulation, flehmen and urination. Besides hiding, moving away or lying-out, non-receptive females fell on their metacarpal joints when approached by a male, a behaviour not yet reported in hippotragine species. This study showed that courtship is the most prominent component of male mating behaviour and that receptivity in females may vary between 1 and 3 days.

Land transformation for anthropogenic use is the leading cause of species decline globally. However, few species are able to succeed in anthropogenically disturbed environments. African clawless otters (Aonyx capensis) occur in a wide variety of habitats, and thus are good model species to investigate animal adaptation to anthropogenic environments. Therefore, we aimed to estimate and compare population densities, group size, and activity time of African clawless otters occurring in a transformed area, Millstream Farm, and two natural areas, Verloren Vallei Nature Reserve and Cobham Nature Reserve. Camera trap arrays, consisting of between 18, 27 and 24 cameras, were placed on all three sites, recording otter presence for a total of 2439 camera days. Using random encounter models, we estimated that the transformed area had the highest density of African clawless otters (8.2 ± 2.3 km−2), compared to otter densities at Verloren Vallei and Cobham Nature Reserve, South Africa, (natural areas) with estimated densities of 0.7 ± 0.2 km−2 and 2.1 ± 0.6 km−2 respectively. We found a significant difference (p = 0.007) between group sizes in the transformed area (detections = 112; group size range = 1–5) and natural areas (detections = 29; group size range = 1–3). Furthermore, there was a significant difference in otter activity time (p = 0.033, activity overlap = 66.5 ± 8.33%) between Verloren Vallei Nature Reserve and the transformed area at Millstream Farm. This study illustrates how African Clawless otters exhibit behavioural plasticity in order to exploit a resource-rich anthropogenic environment.

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...

In a time of increasing environmental change caused by anthropogenic disturbance, there is a greater need to understand animal adaptations to manmade environments. In this regard, the measurement of stress-related endocrine markers provides a useful tool to examine the impact of environmental challenges and the physiological consequences for wildlife occupying such space. The aims of the present study were to validate fecal glucocorticoid metabolite (fGCM) concentrations as a measure of stress using samples from a male African clawless otter (Aonyx capensis; n = 1) and to compare fGCM concentrations of otters occurring in a transformed and in 2 natural areas in South Africa. From the 5 different enzyme-immunoassays (EIA) tested, a cortisol and oxoetiocholanolone (measuring 11,17 dioxoandrostanes) EIA revealed the highest response (74% and 48% increase, respectively) 30 and 24 hours after a stress event (translocation of a captive individual as part of its rehabilitation prior to release), respectively. For both EIAs, fGCM concentrations were comparable for samples collected up to 3 h post-defecation. Using the cortisol EIA for subsequent analyses, fGCM concentrations of animals from the transformed area (n = 20; mean [± SD]: 0.468 [± 0.539] µg/g dry weight [DW]) were significantly higher (P = 0.013) than those from otters in the natural areas (n = 17; 0.242 [± 0.226] µg/g DW). These preliminary results suggest that African clawless otters may have increased adrenocortical activity that could be due to conditions linked to living in a transformed environment.

Poaching is the primary threat to the survival of rhinoceros’ populations. One frequent consequence of poaching is the creation of orphan calves. If found, orphans are taken into captivity for rehabilitation and subsequent release. However, rehabilitation practices can influence their behavior and welfare, potentially compromising their post-release adaptation and survival. In this study, the effects of hands-off and hands-on rehabilitation methods on the behavior, welfare, and adaptation potential of orphaned white rhinoceros (Certatotherium simum simum) were compared. To achieve these aims, 12 behavioral, one physiological, and four physical indicators of welfare and adaptation potential were measured non-invasively on 25 orphaned rhino at two rehabilitation facilities in South Africa. Results indicated that although orphan welfare was not compromised under either rehabilitation method, the hands-off cohort showed fewer indicators of poor welfare and more indicators of good welfare. Regarding adaptation potential, hands-off rehabilitated rhino showed the species’ natural response to humans, and alert and defense behaviors were part of their behavioral repertoire. The hands-on cohort displayed fewer social interactions than the hands-off cohort, showed habituation to humans, and seldom expressed alert or defense behaviors, which could potentially compromise their survival and social integration after release. Post-release studies are required to confirm whether fitness is compromised in hands-on rehabilitated rhino. Until then, we suggest to minimize anthropogenic exposure during rehabilitation in order to maximize welfare and retain crucial behaviors for post-release adaptation and survival.

Social dynamics can play a major role in shaping the population ecology and evolutionary trajectory of a species. This is, for instance, the case in species known to experience infanticide when a dominant male is replaced by another. Infanticide by males has been observed in many taxa, mostly in species that breed year-round and in which a few males monopolize reproduction. In such species, infanticide often occurs after take-overs of breeding groups, and the frequency of take-overs often increases with increasing female group size, therefore raising the risk of infanticide in larger groups. Among ungulates, male infanticide has been reported mostly in harem-forming equids in captive populations, usually following harem-male turnover. In this study, we empirically tested in the wild whether the rate of stallion (i.e. harem-male) turnover increases with the number of reproductive females in harems, and whether these events reduce foal survival. We used a long-term study of wild plains zebra, Equus quagga, in Hwange National Park, Zimbabwe, where, for more than 15 years, stallion turnovers have been recorded and accurate data on pregnancy obtained by hormone assays. We show that stallion turnovers were frequent and, as expected, particularly in harems with many reproductive females. We found that foal survival did not differ significantly between situations when a stallion turnover occurred shortly before or after their birth and when it did not. This result was consistent with the fact that no attacks by stallions on foals of their group or forced matings were observed during the study. Our study offers rare data showing that if male infanticide occurs in wild plains zebra, this should be rare, and has no important consequences for foal survival at the population level.

Males in female-philopatric social groupings leave their natal groups to pursue successive reproductive opportunities in one or more other groups. In vervet monkeys, Chlorocebus pygerythrus , adult males coexist and physical eviction is not a driver of male movement. Migratory decisions are expected to turn on an evaluation of future reproductive opportunity, as indexed principally by local operational sex ratio and relative competitive ability. Although vervet males' reproductive success is correlated with dominance, they are distinctive in that the attainment of rank is contingent on integration into female sociospatial networks and we expect decisions about continued residency to reflect this. We used 8 years' data from three groups to confirm that male dispersal between groups is seasonal in our population, with a peak that is coterminous with androgen levels and precedes peak mating and conception by 4 weeks. The average length of completed residency was 459 days, with an increase in the logged rate of departure after 1428 days, which is 150 days longer than the estimated modal age at first conception by putative daughters. There were positive correlations between a male's initial and highest rank, and between his highest rank and the length of time to reach it. We found that a male's residency was positively and independently associated with his highest achieved rank and both his grooming centrality and proximity degree. Additionally, increasing rank and proximity degree also had positive effects on residency length subsequent to the attainment of his highest rank. The probability of emigration was associated negatively with both female number and grooming centrality scores. We conclude that emigration from a group is linked to male rank attainment and mediated by a male's integration into female sociospatial networks. We found no evidence that emigration preceded the sexual maturity of putative daughters.

Relatively little effort has been directed towards elucidating the role of physiological stress pathways in mediating avian responses to global heating. For free-ranging southern pied babblers, Turdoides bicolor, daily maximum air temperatures (Tmax) between ∼35 and ∼40°C result in reduced foraging efficiency, loss of body mass and compromised breeding success. We tested the hypothesis that very hot days are experienced as stressors by quantifying relationships between Tmax and faecal glucocorticoid metabolite (fGCM) levels in naturally excreted droppings. On days when Tmax38°C, however, fGCM levels increased linearly with Tmax and averaged 190.79±70.13 ng g−1 dry mass. The effects of Tmax on fGCM levels did not carry over to the following morning, suggesting that very hot days are experienced as acute stressors.

Rapid anthropogenic environmental change is expected to impact a host of ecological parameters in Southern Ocean ecosystems. Of critical concern are the consequences of these changes on the range of species that show fidelity to migratory destinations, as philopatry is hypothesized to help or hinder adaptation to climate change depending on the circumstances. Many baleen whales show philopatry to feeding grounds and are also capital breeders that meet migratory and reproductive costs through seasonal energy intake. Southern right whales (Eubalaena australis, SRWs) are capital breeders that have a strong relationship between reproductive output and foraging success. The population dynamics of South Africa's population of SRWs are characterized by two distinct periods: the 1990s, a period of high calving rates; and the late 2010s, a period associated with lowered calving rates. Here we use analyses of stable carbon (δ13C) and nitrogen (δ15N) isotope values from SRW biopsy samples (n = 122) collected during these two distinct periods to investigate foraging ecology of the South African population of SRWs over a time period coincident with the demographic shift. We show that South African SRWs underwent a dramatic northward shift, and diversification, in foraging strategy from 1990s to 2010s. Bayesian mixing model results suggest that during the 1990s, South African SRWs foraged on prey isotopically similar to South Georgia/Islas Georgias del Sur krill. In contrast, in the 2010s, South African SRWs foraged on prey isotopically consistent with the waters of the Subtropical Convergence, Polar Front and Marion Island. We hypothesize that this shift represents a response to changes in preferred habitat or prey, for example, the decrease in abundance and southward range contraction of Antarctic krill. By linking reproductive decline to changing foraging strategies for the first time in SRWs, we show that altering foraging strategies may not be sufficient to adapt to a changing ocean.

The savannah biome supports unique biodiversity and provides a multitude of ecosystem services. Defining background concentrations for trace elements in the environment is beneficial for the determination of nutrient deficiencies/hotspots and for the management of pollution. Sediment and corresponding vegetation samples were collected around 48 surface water points in two savannah wildlife areas for assessment and comparison of 20 trace elements using ICP-MS. Site-specific and matrix-specific differences were evident for essential B, Co, Cu, Fe, Mn, Mo, Ni, Se and Zn, potentially toxic As, Cd, Cr, Hg, Pb and V and additional elements Al, Ba, Sb, Sn and Sr analysed. Sediment and vegetation from all sampled locations at both sites contained single or multiple potentially toxic elements at various concentrations. Although the presence of all elements can be linked to underlying geology and geochemistry specific to each site, evidence of anthropogenic cause was also evident at both sites. This paper covers the widest range of trace elements assessed in protected terrestrial wildlife reserves in the South African savannah biome to date and highlights the potential for deleterious consequences of trace element contamination of the environment.

The southern ground-hornbill (SGH) is a cooperatively breeding bird endemic to eastern and southern Africa, but is endangered in its southern distributional range. The national conservation restoration program harvests redundant chicks for captive breeding and reintroduction; with sexing and social grouping of the species evaluated by throat-skin coloration, with adult males displaying a completely red color compared to dark blue within the red observed in adult females. However, recent findings indicate that dominant and subordinate adult males exhibit patches of blue throat-skin. To optimize SGH management practices, it is vital to determine the role of red and blue coloration, as well as the possible drivers thereof. As a prerequisite, an enzyme immunoassay for monitoring fecal androgen metabolite (fAM) concentrations in SGH was established. Following this, fresh fecal samples were collected from 78 SGH, of various demographics and origin, across 12 captive institutions, to determine whether fAM concentrations differ between blue (B), partially blue (sB), and fully red (R) throat-skin colored SGH. Furthermore, fAM concentrations were compared between males housed in different social groups of different age and sex classes. Individual median fAM concentrations of B, sB, and R adult males did not differ significantly but were considerably higher in B and sB males compared to R males. Social dynamics within captivity, for example, dominance, played no role as a driver of male gonadal activity or throat skin coloration. The results of the study indicate that androgens and apparent social dynamics are not primary determinants of throat coloration in male SGH.

There are few stressful factors which have been investigated to affect adrenocortical function in the African wild dog (AWD) Lycaon pictus. Understanding what animals perceive as stressors is important for not only the implementation of management practices promoting general animal welfare in captivity, but also because a prolonged stress perception is known to disrupt reproduction, immune function and ultimately pose a threat to survival. In this regard, faecal glucocorticoid metabolite (fGCM) measurements are commonly used as a non-invasive approach to assess the impact of factors which may be acting as perceived stressors in wildlife. This study was aimed at determining if there are significant differences in AWD fGCM concentrations as a result of sex, presence of absence of an injury, age-class, body condition, dietary provisioning (through stable nitrogen (δ15N) and carbon (δ13C) isotope analysis), hierarchal status, and setting (i.e. captive or free-ranging). A total of 47 faecal samples were collected immediately post-defaecation from 39 AWDs across four free-ranging sites (Hluhluwe-iMfolozi Park, Madikwe Game Reserve, Kruger National Park and the Waterberg), two permanently captive sites (Johannesburg and Pretoria Zoo), and four temporary captive holding facilities (Hoedspruit Endangered Species Centre and Maremani, Tembe and Zimanga bomas) in South Africa. Captive AWDs had distinctively higher fGCM levels than their free-ranging counterparts, regardless of sex, body condition, hierarchal status, age-class or dietary provisioning. The present study is the first to assess physiological stress responses across permanently captive, temporarily captive and free-ranging AWDs within the South African managed metapopulation, while incorporating the use of stable isotope analysis to quantify differences in dietary isotopic profiles between these different settings. Ultimately this demonstrates the usefulness of fGCM analysis as a tool for assessing animal welfare in both captive and free-ranging AWDs, and underpins the importance of understanding factors perceived as stressors for the management of the species. Keywords: animal welfare, body condition scoring, faecal glucocorticoid metabolite monitoring, managed metapopulation, stress, wildlife management.

Background Glucocorticoids mediate responses to perceived stressors, thereby restoring homeostasis. However, prolonged glucocorticoid elevation may cause homeostatic overload. Using extensive field investigations of banded mongoose (Mungos mungo) groups in northern Botswana, we assessed the influence of reproduction, predation risk, and food limitation on apparent homeostatic overload (n=13 groups, 1542 samples from 268 animals). We experimentally manipulated reproduction and regulated food supply in captive mongooses, and compared their glucocorticoid responses to those obtained from free-living groups. Results At the population level, variation in glucocorticoid levels in free-living mongooses was explained by food limitation: fecal organic matter, recent rainfall, and access to concentrated anthropogenic food resources. Soil macrofauna density and reproductive events explained less and predation risk very little variation in glucocorticoid levels. Reproduction and its associated challenges alone (under regulated feeding conditions) increased glucocorticoid levels 19-fold in a captive group. Among free-living groups, glucocorticoid elevation was seasonal (occurring in late dry season or early wet season when natural food resources were less available), but the timing of peak glucocorticoid production was moderated by access to anthropogenic resources (groups with fewer anthropogenic food sources had peaks earlier in dry seasons). Peak months represented 12- and 16-fold increases in glucocorticoids relative to nadir months with some animals exhibiting 100-fold increases. Relative to the captive group nadir, some free-living groups exhibited 60-fold increases in peak glucocorticoid levels with some animals exhibiting up to 800-fold increases. Most of these animals exhibited 1- to 10-fold increases relative to the captive animal peak. Conclusions Banded mongooses exhibit seasonal chronic glucocorticoid elevation, associated primarily with food limitation and secondarily with reproduction. Magnitude and duration of this elevation suggests that this may be maladaptive for some animals, with possible fitness consequences. In late dry season, this population may face a convergence of stressors (food limitation, agonistic encounters at concentrated food resources, evictions, estrus, mate competition, parturition, and predation pressure on pups), which may induce homeostatic overload.

Non-invasive methods for measuring glucocorticoids and their metabolites are frequently used in ecological, behavioural and physiological studies of mammals. Using faeces, urine and other matrices for such a measurement has considerable advantages in comparison to more traditional methods, but also requires thorough validation of the methods used. Eastern rock sengis (Elephantulus myurus) are fascinating African mammals and the non-invasive monitoring of the adrenocortical activity opens up new opportunities to study their biology. We were able to validate two assays for measuring urinary (uGCM) and faecal glucocorticoid metabolite (fGCM) concentrations in this species using a dose-dependent challenge with adrenocorticotropic hormone (ACTH). A higher concentration of ACTH elicited higher uGCM and fGCM concentrations in both males and females. Interestingly, uGCM and fGCM concentrations and the responses to ACTH were higher in females than in males and small changes in faecal glucocorticoid metabolites could not be reliably detected in males. In contrast to ACTH, a saline injection did not result in an increase in uGCM or fGCM concentrations. The study also provided insight into when responses to a stressor are likely to be detected in the urine and faeces of sengis and opens up new opportunities to study the stress physiology of this and other sengi species. It further emphasises the importance of thoroughly validating non-invasive methods for measuring hormones in both sexes of a species and for incorporating dose-dependent approaches.

The non-invasive monitoring of physiological stress can provide conservation and wildlife managers with an invaluable tool for assessing animal welfare and psychological health of captive and free-ranging populations. A significant decrease in free-ranging primate populations globally and an increase in captive-housed primates have led to a need to monitor the stress and general welfare of these animals. We examined the suitability of three enzyme immunoassays (EIAs) for monitoring stress-related physiological responses in the samango monkey, Cercopithecus albogularis erythrarchus. We conducted an adrenocorticotropic hormone (ACTH) challenge on a male and female at the National Zoological Garden, Pretoria, South Africa. Individual faecal samples were collected 8 days pre- and post-ACTH administration and subsequently analysed for faecal glucocorticoid metabolite (fGCM) concentrations. During the study, biological stressors occurred for both the male and female. Two of the three EIAs tested (11-oxoetiocholanolone I and II) were able to reliably monitor fGCM alterations throughout the study period in both sexes. The 11-oxoetiocholanolone I EIA, however, had the lowest mean deviation from the calculated baseline value and was thus chosen as the preferred assay. Both the physiological activation of the stress response and the biological response to a stressor could be monitored with the chosen assay. The successful establishment of a reliable, non-invasive method for monitoring adrenocortical activity in C. albogularis erythrarchus will now allow conservationists, scientific researchers and wildlife managers to evaluate the level of stress experienced, and general welfare, by animals in captivity as well as free-ranging populations.

Understanding the reproductive biology of the roan antelope (Hippotragus equinus) (É. Geoffroy Saint-Hilaire, 1803) is crucial to optimise breeding success in captive breeding programmes of this threatened species. In this study, the pattern of faecal androgen metabolite (fAM) production related to reproductive events (calving or birthing, mating, gestation, and lactation), sexual behaviours as well as environmental cues were studied in captive adult male roan antelope. Faecal sample collection and behavioural observations were carried out from August 2017 to July 2018 for three reproductive males participating in a conservation breeding programme at the Lapalala Wilderness Nature Reserve in South Africa. As a prerequisite, the enzyme immunoassay used in this study was biologically validated for the species by demonstrating a significant difference between fAM concentrations in non-breeding adults, breeding adults and juvenile males. Results revealed that in adults males, the overall mean fAM levels were 73% higher during the breeding period compared to the non-breeding periods, and 85% higher when exclusively compared to the lactation/gestation periods, but only 5.3% higher when compared to the birthing period. Simultaneously, fAM concentrations were lower during the wet season compared to the dry season, increasing with a reduction in photoperiod. With the exception of courtship, frequencies of sexual behaviours monitored changed in accordance with individual mean fAM concentrations in male roan antelope, the findings suggest that androgen production varies with the occurrence of mating activity and may be influenced by photoperiod but not with rainfall.

Dominant females of cooperative breeding species often use aggression to suppress reproduction of subordinate females, resulting in subordinates experiencing stress-related increases in glucocorticoid levels, which may cause reproductive down-regulation. This would suggest a general pattern with higher glucocorticoid levels in subordinate compared to dominant individuals; however, the opposite was found in a number of cooperatively breeding species. Furthermore, breeding females of the cooperatively breeding Damaraland mole-rats (Fukomys damarensis) exhibit very high androgen concentrations during the wet season, presumably to support their breeding monopoly. Hormone analysis in Damaraland mole-rats have typically been measured using plasma and urine, but faecal analysis offers additional advantages especially for field studies on this species. The present study examines the suitability of Damaraland mole-rat faecal samples for determining glucocorticoid metabolite (fGCM) and androgen metabolite (fAM) concentrations using enzyme immunoassays. Using these assays, we further evaluated the effects of breeding status on fGCM and fAM concentrations in wild-caught and captive Damaraland mole-rats. Wild-caught breeding and non-breeding males and females exhibited no differences in fAM concentrations. Immunoreactive fGCM concentrations were only high in male breeders and comparatively low in non-breeders and breeding females. Concentrations of fAMs and fGCMs were similar in captive males and females, but fAM concentrations were elevated in captive compared to wild-caught individuals, which may be related to a higher reproductive activity due to removal from the breeding female. The relatively uniform fAM and fGCM concentrations found in wild-caught mole-rats may be explained by a stable colony structure during the dry season during which this study was conducted. Limited dispersal opportunities result in lower aggression and stress levels within a colony and as a result lower fAM and fGCM concentrations.

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.

Photoperiod and food availability often interact in the regulation of reproductive development in seasonal environments. We tested the effects of a 10% food restriction and short- (SD) and long-day (LD) photoperiods on reproductive preparedness of male spiny mice (Acomys spinosissimus) from South Africa. Males were either fed ad libitum or a 10% restricted diet and on both food regimes, they were exposed to either a 14 h light:10 h dark (LD) or 10 h light:14 h dark (SD) photoperiod. Testes size, seminiferous tubule diameter and faecal androgen metabolite (fAM) concentrations were larger in food restricted males subjected to LD than in any of the other groups. SD photoperiods did not suppress reproductive development, despite earlier findings, which may be explained by “spontaneous” recrudescence due to prolonged exposure to SD. The increased development of the gonads and higher fAM concentrations observed in food restricted LD males may indicate a terminal investment strategy in male spiny mice with males investing more into reproduction in the face of reduced future survival.

The National Research Foundation, South Africa and a postgraduate bursary from the DST-NRF SARChI chair for Mammal behavioural ecology and physiology to (NCB).

One of the primary sources of perceived stress is the social environment of an animal and the interactions with conspecifics. An essential component of the response to a stressor is the activation of the hypothalamic-pituitary-adrenocortical axis, which results amongst others in a temporal increase in circulating glucocorticoid (GC) levels. Giraffes occur in a highly flexible fission-fusion social system and group compositions can change on a daily basis, with bulls establishing an age-related dominance hierarchy and showing a roaming strategy in the search for fertile females. The aim of this study was to non-invasively monitor the influence of different group compositions (mixed sex groups vs. all-male groups) on GC concentrations in free ranging giraffe bulls of different age classes. We collected fecal samples from free-ranging giraffe bulls for 12months in a South African Private Game Reserve to examine age- and social context-related patterns of fecal GC metabolite (fGCM) concentrations. We found that fGCM levels in giraffe bulls are age-class dependent, as well asassociated with changes in the social environment. Independently of the social setting, bulls of the youngest age class exhibited the highest fGCM levels compared to bulls of the other two older age-classes, with differences most pronounced when the bulls are associated in all-male groups. In contrast, an almost reversed picture appears when looking at the fGCM levels of sexually active individuals in mixed sex groups, where highest levels were found for the bulls in the oldest age-class, and the lowest for the bulls in the youngest age-class. The study stresses the importance to taking factors such asage-related status and social settings into account, when interpreting fGCM levels in free ranging giraffes.

A physiological stress response can be triggered by a variety of intrinsic and extrinsic stimuli, but minimal information is available about the physiological stress response related to pain in wildlife. Recently established techniques now allow the non-invasive measurement of faecal glucocorticoid metabolite (fGCM) concentrations to monitor the physiological stress response in giraffe. We examined the consequences of injury of various severities and loss of body condition in relation to glucocorticoid output in free-ranging giraffes. Body condition (BC) was visually estimated based on the amount of fat and muscle covering the bones, using a species-specific scoring system (one – emaciated to eight – obese). An adapted animal trauma triage scoring system was also applied to evaluate the severity of injuries observed. Individual fGCM concentrations were determined to assess stress-related glucocorticoid output using an enzyme immunoassay technique, and compared with assigned BC and injury scoring. Significantly elevated fGCM levels were found in injured individuals that showed wounds with deep tissue involvement and/or poor BC, but not in individuals that showed superficial wounds. Responsible for the observed changes in fGCM levels may be a combination of differences in the severity of the injuries and the subsequent degree of pain associated with it, the influence of the stress response on the energetic condition, and the duration of the injuries. The results of this study are somewhat limited due to the small sample size, and therefore the effect of food intake on the body conditions cannot be controlled for. However, euthanasia is a common management tool used to prevent unnecessary suffering, nevertheless, especially in wild animals the severity of an injury and the associated pain perceived may be difficult to assess. Combining an assessment of BC and analysis of individual stress-hormone levels can help improve health assessments in free-ranging giraffes and thus assist management decisions.

Non-invasive techniques for the monitoring of animal well-being, such as faecal hormone analysis, are increasingly becoming some of the most desirable methods for addressing practical conservation questions. Despite the widespread use of faecal hormone measurements for monitoring responses to stressors and its known applicability to African wild dogs Lycaon pictus, the potential influence of time of collection post-defaecation on stress-steroid concentrations in faecal matter has not yet been investigated. In the present study, we determined the rate at which African wild dog faecal glucocorticoid metabolite (fGCM) concentrations change over a 16-day period post-defaecation, in order to provide recommendations for best sampling practice. No significant changes in fGCM concentrations were found for the first 48 h post-defaecation. However, an approximately 30% increase in fGCM concentrations were already noted between day 1 and day 2, followed by a significant 150% increase at 96 h post-defaecation. We therefore suggest that respective faecal material should be collected within the first 24 h post-defaecation to ensure the reliability of fGCM analysis. In addition, we collected baseline data denoting the fGCM concentrations of captive African wild dogs sampled across three South African captive sites. Determined baseline fGCM concentrations differed between African wild dogs at the sites sampled. These data could be used in future studies aimed at identifying the key stressor complexes perceived by captive African wild dogs in order to improve management strategies.

Living under predation risk may alter both behaviour and physiology of potential prey. In extreme cases, such alterations may have serious demographic consequences, and recent studies support that non-lethal effects of predation may have broad ecological consequences. However, behavioural and physiological responses to predation risk may be related to trade-offs associated with resource acquisition and direct predation risk. We validated an enzyme-linked immunoassay (EIA) for non-invasive monitoring of stress in plains zebras (Equus quagga) from faecal material. We used this assay in combination with behavioural data to assess if plains zebras living with and without lions (Panthera leo) in a mountain savannah in southern Africa differed in behaviour and physiology, and if such differences were influenced by seasons with contrasting resource availability. Zebra group sizes did not differ between areas with and without lions, but zebra groups had more juveniles in an area with lions than groups in an area without lions, but only during the wet season. Similarly, we observed differences in individual vigilance, foraging behaviour and stress hormone concentrations, but all these differences were influenced by seasons. Despite these seasonal influences, our study did not suggest that zebras in an area with lions spent a higher proportion of time being vigilant, a lower proportion of time foraging, or had higher stress hormone levels. Our results instead suggest that zebras' responses to lion presence were highly context dependent and the result of complex interactions between resource abundance and cues about predation risk. Because of the obvious ecological and evolutionary ramifications of such findings, we argue that further research is needed to define the spatial and temporal scales over which predators impose indirect effects on their prey.

The golden langur (Trachypithecus geei) is an endangered primate endemic to northern India and Bhutan. The main stressors to the species are habitat degradation and fragmentation. Non-invasive fecal glucocorticoid metabolite (fGCM) analysis is a powerful tool for assessing stress associated with environmental disturbances in wildlife. However, interspecific differences in glucocorticoid metabolism require careful selection of the antibody used in their quantification. The goals of this study were to: 1) validate an enzyme immunoassay (EIA) to determine fecal GC metabolite (fGCM) concentrations in the golden langur and 2) compare fGCM concentrations between golden langurs living under different environmental conditions. We compared five enzyme immunoassays for determining stress related physiological responses in golden langurs by performing an adrenocorticotropic stimulation test in both sexes. Our validations identified an 11-oxoaetiocholanolone EIA detecting 11, 17 dioxoandrostanes as the most suitable assay for monitoring adrenocortical activity in the species. FGCM concentrations from semi-captive temple langurs were significantly higher than respective stress steroid concentrations in free-ranging or pet/zoo langurs. This study presents a validated practical method for non-invasive monitoring of adrenocortical function in this rare and declining species.

Stress in animals has been linked to behavioural and physiological changes in response to environmental, social and anthropogenic stimuli. Hence, stress-related responses in animals, especially in rodents, have been used as biological indicators of ecosystem health. This study aimed to establish an enzyme immunoassay (EIA) for monitoring adrenocortical activity in free-ranging Namaqua rock mice Micaelamys namaquensis (Rodentia: Muridae) using faeces as a prerequisite for assessing the effects of anthropogenic land use and season on faecal glucocorticoid metabolite (fGCM) concentration. Rodents were live-trapped seasonally across four land use types: an agricultural crop farm, an agricultural livestock farm, a human-populated site and a nature reserve; all situated in Limpopo Province, South Africa. Determined fGCM concentrations from capture and recapture events were used for biologically validating an EIA detecting steroids with a 5α-3β-11β-diol structure. Recapturing resulted in a significant overall 40% elevation of individual fGCM concentrations demonstrating the effectiveness of the chosen EIA to reliably detect glucocorticoid output in the study species. Neither land use type nor season affected fGCM concentrations in the species, suggesting that land use and season-related environmental changes do not necessarily act as stressors for M. namaquensis, presumably due to their adaptive and resilient nature. Such species can be used to identify ecosystems affected by human-mediated disturbances and allow insights into the management and restoration of these threatened ecosystems and their associated species.

The phenotype of parents can have long-lasting effects on the development of offspring as well as on their behaviour, physiology, and morphology as adults. In some cases, these changes may increase offspring fitness but, in others, they can elevate parental fitness at a cost to the fitness of their offspring. Here, we show that in Kalahari meerkats (Suricata suricatta) the circulating glucocorticoid hormones of pregnant females affect the growth and cooperative behaviour of their offspring. We performed a 3-year experiment in wild meerkats to test the hypothesis that stress-mediated maternal effects reduce the potential for offspring to reproduce directly and therefore cause them to exhibit more cooperative behaviour. Daughters (but not sons) born to mothers treated with cortisol during pregnancy were smaller early in life and exhibited significantly more of two types of cooperative behaviour (pup rearing and feeding) once they were adults compared to offspring from control mothers. They also had lower measures of stress hormones as they aged, which could explain the observed increases in cooperative behaviour. Because early growth is a crucial determinant of fitness in female meerkats, our results indicate that stress-mediated maternal effects may reduce the fitness of offspring but may elevate parental fitness as a consequence of increasing the cooperative behaviour of their daughters. Significance Statement Maternal characteristics can have a profound influence on offspring, which are referred to as maternal effects. It is widely assumed that selfish maternal effects, where mothers modify offspring to benefit their own fitness and decrease the fitness of offspring, are rare but this assumption is rarely tested. We show that a stress-mediated maternal effect in meerkats altered the growth, behaviour, and stress physiology of daughters in such a way that benefitted mothers at the expense of their daughters. Daughters from stressed mothers grew more slowly early in life, which should reduce their future reproductive opportunities, but exhibited more cooperative behaviour that should directly benefit their mother. This highlights the important role of sociality in the evolution of maternal effects.

Amphibians are experiencing drastic population declines. New semi-invasive conservation methods are required to protect threatened species. The collection of mucus, from dorsal and ventral body regions, is an ideal matrix for monitoring stress hormones, and thus physiological stress, in African amphibians. This technique will assist in amphibian conservation in Africa., The complex interaction between factors leading to amphibian declines is responsible for the inability to develop robust, standardized conservation action plans. Monitoring physiological stress responses in amphibians may provide an ideal tool to assist conservationists in this regard. This study aimed to validate dermal secretions as a robust matrix for monitoring glucocorticoid alterations in an African amphibian, the edible bullfrog (Pyxicephalus edulis). Both a biological (handling) and physiological (ACTH challenge) validation were conducted to determine which of five available enzyme immunoassays (EIAs) tested is most suited for monitoring alterations in dermal glucocorticoid (dGC) concentrations. Additionally, the most optimal body region for monitoring dGC concentrations in P. edulis was identified. To confirm the activation of the hypothalamic–pituitary–interrenal (HPI) axis following ACTH administration, urinary glucocorticoid metabolites (uGCM) were also quantified. The tested corticosterone EIA was the only assay able to monitor alterations in dGC concentrations following the handling event in P. edulis. Further validation during the ACTH challenge supported the corticosterone EIA in this regard. Dermal secretions collected from both the ventral and dorsal regions were suitable for monitoring dGC concentrations in both sexes. A considerable increase in uGCM concentrations following ACTH administration was found, suggesting that the dGC concentrations observed were derived from the HPI axis. This study offers further support for the use of dermal secretions as an important matrix for monitoring physiological stress, and thus general health, in amphibian species.

Tigers continue to face unprecedented threats to their existence due to poaching, habitat loss, habitat fragmentation and anthropogenic disturbances. The present study examines the physiological stress response of tigers due to anthropogenic activities including wildlife tourism in Bandhavgarh Tiger Reserve and Kanha Tiger Reserve using faecal glucocorticoid metabolite (fGCM) measurement. We collected a total of 341 faecal samples from both reserves during tourism and non-tourism periods. Data on various anthropogenic disturbances including tourism activities like number of vehicles and visitors were also collected. We ascertained the species identity and sex of all the samples collected using genetic markers. fGCMs were extracted using a previously reported procedure, and fGCM concentrations were subsequently determined using an established enzyme immunoassay. There was no significant difference in overall mean fGCM concentrations between the two tiger reserves, but within each reserve, concentrations were significantly higher in tigers during the tourism period as compared to the non-tourism period. We also found that the number of tourist vehicles and disturbance level significantly correlated with fGCM concentrations. This study further supports the assumption that unbridled tourism associated with high anthropogenic disturbance can be related to perceived stress and consequently may have an impact on the reproductive fitness of tigers and long-term survival of isolated populations.

Hormone monitoring in keratinous tissues has become increasingly popular. The insoluble keratin materials are generally pulverised before hormone extraction; however, this is difficult for thicker keratin structures like baleen plates or hooves. A new method, involving the use of keratinase, allows enzymatic digestion of keratin and hormone analysis in the resulting suspension. Pangolins are unique mammals covered in keratinous scales, which are one of the reasons these animals are extensively trafficked. This study aimed to investigate the suitability of Temminck's pangolin scales as hormone matrix for quantifying reproductive steroids. A protocol was developed to digest scales with a keratinase before measuring hormone concentrations. This method can be used to investigate the reproductive endocrinology of Temminck's pangolins but may also be extended to the other extant pangolin species.•Keratinase digests Temminck's pangolin scales and reproductive steroid metabolite concentrations are measurable in the resulting suspension.•Isopropanol is an ideal washing solvent for scales to remove surface contaminants and scale sample mass should be standardised to allow comparisons.•Any section of a scale and scales from any pangolin body region can be used as samples for hormone quantification., Graphical abstract Image, graphical abstract

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

The use of carbon dioxide (CO2) exposure as a means of animal euthanasia has received considerable attention in mammals and birds but remains virtually untested in reptiles. We measured the behavioral responses of four squamate reptile species (Homalopsis buccata, Malayopython reticulatus, Python bivitattus, and Varanus salvator) to exposure to 99.5% CO2 for durations of 15, 30, or 90 minutes. We also examined alterations in plasma corticosterone levels of M. reticulatus and V. salvator before and after 15 minutes of CO2 exposure relative to control individuals. The four reptile taxa showed consistent behavioral responses to CO2 exposure characterized by gaping and minor movements. The time taken to lose responsiveness to stimuli and cessation of movements varied between 240-4260 seconds (4-71 minutes), with considerable intra- and inter-specific variation. Duration of CO2 exposure influenced the likelihood of recovery, which also varied among species (e.g., from 0-100% recovery after 30-min exposure). Plasma corticosterone concentrations increased after CO2 exposure in both V. salvator (18%) and M. reticulatus (14%), but only significantly in the former species. Based on our results, CO2 appears to be a mild stressor for reptiles, but the relatively minor responses to CO2 suggest it may not cause considerable distress or pain. However, our results are preliminary, and further testing is required to understand optimal CO2 delivery mechanisms and interspecific responses to CO2 exposure before endorsing this method for reptile euthanasia.

Fynbos is a unique endemic vegetation type belonging to the Cape Floral Kingdom in the Western Cape Province of South Africa, representing the smallest of the six floral kingdoms in the world. Nowadays, only a few game reserves in this region support populations of African elephants (Loxodonta africana), and thus, little information exists regarding the suitability of the nutritionally poor Fynbos vegetation for these megaherbivores. Using already established non-invasive methods, the monitoring of individual body conditions and fecal glucocorticoid metabolite (fGCM) concentrations, as a measure of physiological stress, was performed to examine a herd of 13 elephants in a Western Cape Province Private Game Reserve, during two monitoring periods (April and June 2018), following a severe drought. The results indicate that overall median body condition scores (April and June: 3.0, range 2.0&ndash, 3.0) and fGCM concentrations (April: 0.46 &micro, g/g dry weight (DW), range 0.35&ndash, 0.66 &micro, g/g DW, June: 0.61 &micro, g/g DW, range 0.22&ndash, 1.06 &micro, g/g DW) were comparable to those of other elephant populations previously studied utilizing the same techniques. These findings indicate that the individuals obtain sufficient nutrients from the surrounding Fynbos vegetation during the months monitored. However, a frequent assessment of body conditions and stress-associated fGCM concentrations in these animals would assist conservation management authorities and animal welfare practitioners in determining ways to manage this species in environments with comparably poorer nutritional vegetation.

Developing non-invasive techniques for monitoring physiological stress responses has been conducted in a number of mammal and bird species, revolutionizing field-based endocrinology and conservation practices. However, studies validating and monitoring glucocorticoid concentrations in reptiles are still limited. The aim of the study was to validate a method for monitoring glucocorticoid metabolite concentrations in urine (uGCM) and faeces (fGCM) of the cordylid lizard, the Sungazer (Smaug giganteus). An adrenocorticotropic hormone (ACTH) challenge was conducted on one male and two females with both urine and faecal material being collected during baseline and post-injection periods. Steroid extracts were analysed with four enzyme immunoassays (EIAs)namely: 11-oxoaetiocholanolone, 5α-pregnane-3β-11β-21-triol-20-one, tetrahydrocorticosterone, and corticosterone. A considerable response in fGCM and uGCM concentrations following ACTH administration was observed in all subjects, with the 5α-pregnane-3β-11β-21-triol-20-one and tetrahydrocorticosterone EIAs appearing to be the most suited for monitoring alterations in glucocorticoid metabolite concentrations in S. giganteus using faeces or urine as hormone matrix. Both EIAs showed a significantly higher concentration of glucocorticoid metabolites in faeces compared to urine for both sexes. Collectively, the findings of this study confirmed that both urine and faeces can be used to non-invasively assess adrenocortical function in S. giganteus.

Due to the strong individual cost of being predated, potential prey species alter their behavior and physiology in response to predation risk. Such alterations may cause major indirect consequences on prey populations that are additive to the direct demographic effects caused by prey being killed. However, although earlier studies showed strong general effects of the presence of apex predators, recent data suggest that indirect effects may be highly context dependent and not consistently present. We combined behavioral data with data on endocrine stress and stable isotopes to assess landscape level effects of lion (Panthera leo) presence on two prey species in South Africa, impala (Aepyceros melampus) and blue wildebeest (Connochaetes taurinus). We also evaluated if there was any seasonal variation in such effects. In addition, we provide results from a physiological validation for an enzyme-linked immunoassay (EIA) that can be used for non-invasive monitoring of glucocorticoid stress metabolite concentrations in impala from fecal pellets. We did not find any significant differences in vigilance behavior, fecal glucocorticoid metabolite concentrations, δ13C values or isotope niche breadth between animals living with and without lions for either species. However, wildebeest living in a reserve with lions spent more time foraging compared to wildebeest in a lion-free environment, but only during the wet season. Values of fecal δ15N suggest a shift in habitat use, with impala and wildebeest living with lions potentially feeding in less productive areas compared to animals living without lions. For both species, characteristics of the social groups appeared to be more important than individual characteristics for both foraging and vigilance behavior. Our results highlight that antipredator responses may be highly dynamic and scale-dependent. We urge for further studies that quantify at what temporal and spatial scales predation risk is causing indirect effects on prey populations.

Background: Faecal hormone metabolite measurement is a widely used tool for monitoring reproductive function and response to stressors in wildlife. Despite many advantages of this technique, the delay between defaecation, sample collection and processing may influence steroid concentrations, as faecal bacterial enzymes can alter steroid composition post-defaecation. Objectives: This study investigated changes in faecal glucocorticoid (fGCM), androgen (fAM) and progestagen (fPM) metabolite concentrations in faeces of a male and female African elephant ( Loxodonta africana ) post-defaecation and the influence of different faeces-drying regimes. Method: Subsamples of fresh faeces were frozen after being dried in direct sun or shade for 6, 20, 24, 48 and 72 h and 7 and 34 days. A subset of samples for each sex was immediately frozen as controls. Faecal hormone metabolite concentrations were determined using enzyme immunoassays established for fGCM, fAM and fPM monitoring in male and female African elephants. Results: Hormone metabolite concentrations of all three steroid classes were stable at first, but changed distinctively after 20 h post-defaecation, with fGCM concentrations decreasing over time and fPM and fAM concentrations steadily increasing. In freeze-dried faeces fGCM concentrations were significantly higher than respective concentrations in sun-dried material, which were in turn significantly higher than fGCM concentrations in shade-dried material. In contrast, fAM concentrations were significantly higher in sun- and shade-dried faeces compared to freeze-dried faeces. Higher fPM concentrations were also found in air-dried samples compared to lyophilised faeces, but the effect was only significant for sun-dried material. Conclusion: The revealed time restriction for collecting faecal material for hormone monitoring from elephants in the wild should be taken into account to assure reliable and comparable results. However, if logistics allow a timely collection, non-invasive hormone measurement remains a powerful and reliable approach to provide information about an elephant’s endocrine status.

Steroid hormones play an important role in female reproductive physiology and behaviour and are often used to monitor important female reproductive events. However, such studies are often attempted on captive populations alone, delivering limited data. One such example is the African lesser bushbaby, Galago moholi, for which contradicting observational data exist between captive and free-ranging populations, while hormonal analyses have only been obtained from a single captive population. To extend and rectify the limited information, we monitored faecal progestagen and oestrogen metabolite levels across various important life history stages of both captive and free-ranging G. moholi. We additionally recorded changes in vaginal state as well as the occurrence of reproductive and aggressive behaviour throughout the study. Data from our captive population revealed an ovarian cycle length of 33.44 ± 0.59 days (mean ± SD), with follicular and luteal phases of 14.2 ± 1.0 and 19.1 ± 1.5 days, respectively, and an average pregnancy length of 128 ± 3.3 days. The initiation of female reproductive activity was closely linked to an oestrus-related increase in faecal oestrogen metabolite levels. Four of the seven captive females monitored in our study conceived during the May mating period, with one additional female fertilised in September, supporting the idea that the September mating period functions as a back-up for female G. moholi. Identified benchmark faecal progestagen metabolite levels (non-pregnant: >1 µg/g dry weight (DW), pregnant: >9 µg/g DW) should help researchers to determine pregnancy status of randomly wild-caught females in even a cross-sectional study setup.

In cooperative breeders, aggression from dominant breeders directed at subordinates may raise subordinate stress hormone (glucocorticoid) concentrations. This may benefit dominants by suppressing subordinate reproduction but it is uncertain whether aggression from dominants can elevate subordinate cooperative behaviour, or how resulting changes in subordinate glucocorticoid concentrations affect their cooperative behaviour. We show here that the effects of manipulating glucocorticoid concentrations in wild meerkats (Suricata suricatta) on cooperative behaviour varied between cooperative activities as well as between the sexes. Subordinates of both sexes treated with a glucocorticoid receptor antagonist (mifepristone) exhibited significantly more pup protection behaviour (babysitting) compared to those treated with glucocorticoids (cortisol) or controls. Females treated with mifepristone had a higher probability of exhibiting pup food provisioning (pup-feeding) compared to those treated with cortisol. In males, there were no treatment effects on the probability of pup-feeding, but those treated with cortisol gave a higher proportion of the food they found to pups than those treated with mifepristone. Using 19 years of behavioural data, we also show that dominant females did not increase the frequency with which they directed aggression at subordinates at times when the need for assistance was highest. Our results suggest that it is unlikely that dominant females manipulate the cooperative behaviour of subordinates through the effects of aggression on their glucocorticoid levels and that the function of aggression directed at subordinates is probably to reduce the probability they will breed.

In many mammal species, androgen levels in males are elevated during periods of mating activity, often to facilitate aggressive behavior between males over access to fertile females. However, this pattern might be less obvious in species with a rather low male-male aggression rate, or in those that are not strictly seasonal breeders. A complex social structure, as well as additional social and environmental factors, might add more to the complexity. Here, we applied a non-invasive method to monitor fecal androgen metabolite (fAM) levels in free-ranging giraffe bulls over a period of months to examine longitudinal patterns of androgen metabolite concentrations in relation to observed male sexual behavior in different age classes. Giraffes are non-seasonal breeders, living in a fission-fusion social system and males show a roaming strategy to search for fertile females. Our results show that season has an impact on fAM levels in free-ranging giraffes, with respective steroid concentrations being higher in summer. In the presence of females, fAM levels of bulls are significantly higher compared to when found in all-male groups, with old adult bulls showing the highest fAM levels. In contrast, young adult bulls have overall slightly higher fAM levels compared to old adult bulls when residing in all male groups. Sexual behavior increases fAM levels only in old adult bulls.

Mammalian parental investment (i.e. care of descendant offspring) is largely biased towards maternal contributions due to the specific feeding needs of mammalian offspring; however, varying degrees of paternal investment have been reported in about 10% of all mammalian species. Within the order Carnivora, paternal contribution to rearing offspring is particularly high: an estimated 32% of all studied carnivore species exhibit direct paternal care. Despite the prominence of paternal investment in carnivores, the endocrine basis of this behaviour is not well understood. This review examines the current – highly constrained – state of knowledge about the endocrine basis of carnivore paternal investment. We attempt to link changes in androgen and glucocorticoid levels with variation in direct and indirect paternal care behaviour making specific predictions regarding the way forward. Well-studied species, such as bat-eared foxes (Otocyon megalotis), dwarf mongoose (Helogale parvula) and meerkats (Suricata suricatta), where social dynamics are relatively well understood, can act as ideal model systems through which we may further investigate the endocrine basis of paternal investment in carnivores.Keywords: androgens, carnivore, glucocorticoids, paternal investment

Rhinoceros populations in Africa are under severe threat as a result of surging poaching rates and risk-mitigation strategies are continuously adapted in an attempt to ensure the survival of the species. This study compared faecal glucocorticoid metabolite (fGCM) levels of two age classes of limited free-ranging female white rhinos with fGCM levels of adult free-ranging female white rhinos. Subsequently, fGCM alterations in the limited free-ranging animals were monitored following routine dehorning as a measure of the animals’ short-term physiological stress response. Baseline fGCM levels differed significantly between tested groups, with both free-ranging and limited free-ranging adult animals showing significantly higher fGCM levels compared with limited free-ranging juvenile females. In contrast, baseline fGCM levels did not differ significantly between limited free-ranging and free-ranging adult individuals. Routine dehorning procedures resulted in a short-term stress response expressed by a significant increase in fGCM levels 48 h post-dehorning, with stress steroid levels returning to pre-dehorning concentrations 72 h after the procedure.Keywords: faecal glucocorticoid metabolites, non-invasive hormone monitoring, physiological stress, South Africa

It is well established that animal vocalizations can encode information regarding a sender’s identity, sex, age, body size, social rank and group membership. However, the association between physiological parameters, particularly stress hormone levels, and vocal behavior is still not well understood. The cooperatively breeding African meerkats (Suricata suricatta) live in family groups with despotic social hierarchies. During foraging, individuals emit close calls that help maintain group cohesion. These contact calls are acoustically distinctive and variable in rate across individuals, yet, information on which factors influence close calling behavior is missing. The aim of this study was to identify proximate factors that influence variation in call rate and acoustic structure of meerkat close calls. Specifically, we investigated whether close calling behavior is associated with sex, age and rank, or stress hormone output (i.e., measured as fecal glucocorticoid metabolite (fGCM) concentrations) as individual traits of the caller, as well as with environmental conditions (weather) and reproductive seasonality. To disentangle the effects of these factors on vocal behavior, we analyzed sound recordings and assessed fGCM concentrations in 64 wild but habituated meerkats from 9 groups during the reproductive and non-reproductive seasons. Dominant females and one-year old males called at significantly higher rates compared to other social categories during the reproductive season. Additionally, dominant females produced close calls with the lowest mean fundamental frequencies (F0) and the longest mean pulse durations. Windy conditions were associated with significantly higher call rates during the non-reproductive season. Fecal GCM concentrations were unrelated to close calling behavior. Our findings suggest that meerkat close calling behavior conveys information regarding the sex and social category of the caller, but shows no association with fGCM concentrations. The change in call rate in response to variation in the social and ecological environments individuals experience indicates some degree of flexibility in vocal production. peerReviewed

In mammals, glucocorticoid (i.e. GC) levels have been associated with specific life-history stages and transitions, reproductive strategies, and a plethora of behaviors. Assessment of adrenocortical activity via measurement of glucocorticoid metabolites in feces (FGCM) has greatly facilitated data collection from wild animals, due to its non-invasive nature, and thus has become an established tool in behavioral ecology and conservation biology. The aim of our study was to validate a fecal glucocorticoid assay for assessing adrenocortical activity in meerkats (Suricata suricatta), by comparing the suitability of three GC enzyme immunoassays (corticosterone, 11β-hydroxyetiocholanolone and 11oxo-etiocholanolone) in detecting FGCM increases in adult males and females following a pharmacological challenge with adrenocorticotropic hormone (ACTH) and biological stimuli. In addition, we investigated the time course characterizing FGCM excretion, the effect of age, sex and time of day on FGCM levels and assessed the potential effects of soil contamination (sand) on FGCM patterns. Our results show that the group specific 11β-hydroxyetiocholanolone assay was most sensitive to FGCM alterations, detecting significant and most distinctive elevations in FGCM levels around 25 h after ACTH administration. We found no age and sex differences in basal FGCM or on peak response levels to ACTH, but a marked diurnal pattern, with FGCM levels being substantially higher in the morning than later during the day. Soil contamination did not significantly affect FGCM patterns. Our results emphasize the importance of conducting assay validations to characterize species-specific endocrine excretion patterns, a crucial step to all animal endocrinology studies using a non-invasive approach. peerReviewed

The giraffe (Giraffa camelopardalis) is usually described as an exclusive browser, feeding only on shrubs and trees, preferrably between 2 and 5 m above ground (Lamprey, 1963; McNaughton & Georgiadis, 1986; Ciofolo & Le Pendu, 2002). Although browsing seems to be an easier form of feeding for giraffes in terms of accessibility and vigilance (Young & Isbell, 1991), a few studies mention that the giraffe also ‘very occasionally’ feeds on grass (Pienaar, 1963; Du Toit, 2005). To be able to graze, a giraffe has to adopt the typical ‘drinking position’, where the forelegs are splayed out laterally, and sometimes the carpal joints are also flexed. In this position, the animals are particularly vulnerable to predators (Periquet et al., 2010). In this note, we show that grazing, although not a core activity, is a recurrent event in a nutrient-poor environment such as Hwange National Park and suggest a possible function.

International audience; Regulative endocrine mechanisms influence the reproductive behaviour and success of mammals, but they have been studied predominantly in domestic and captive animals. The study aims at describing the pattern of faecal 20-oxopregnane and oestrogen concentrations during pregnancy in wild plains zebra Equus quagga chapmani. Data were collected during wet and dry seasons 2007-2009. Enzyme Immunoassays were used to determine 20-oxopregnane and oestrogen concentrations in faecal samples (n=74) collected from individual mares (n=32) whose dates of foaling were known through long-term monitoring. Hormonal profiles were described with a General Additive Model (GAM: Hormone ∼ Days to Foaling). Faecal 20-oxopregnanes have a complex cycle during pregnancy (GAM, n=70, R(2)=0.616, p200ng/g DW) of faecal 20-oxopregnanes associated with high (>160ng/g DW) faecal oestrogen levels indicate mid-pregnancy in c.90% of cases (16/17). High faecal 20-oxopregnanes (>200ng/g DW) and low faecal oestrogen levels (

The social organization of giraffes ( Giraffa camelopardalis ) imposes a high-cost reproductive strategy on bulls, which adopt a ‘roving male’ tactic. Our observations on wild giraffes confirm that bulls indeed have unsynchronized rut-like periods, not unlike another tropical megaherbivore, the elephant, but on a much shorter timescale. We found profound changes in male sexual and social activities at the scale of about two weeks. This so far undescribed rutting behaviour is closely correlated with changes in androgen concentrations and appears to be driven by them. The short time scale of the changes in sexual and social activity may explain why dominance and reproductive status in male giraffe in the field seem to be unstable.

International audience; Mammalian reproductive rates vary among individuals for physiological and environmental reasons. This study aims to determine reproductive rates from an individually monitored population of wild Plains zebras Equus quagga, and to assess the sources of variability in inter-birth intervals. The animals were monitored, where possible, every six months from 2004 to 2011. Thirty nine intervals corresponding to 65 births in 26 mares were identified, using direct observations and faecal steroid monitoring. Mean foaling rate of the population is 0.74 foal/year, and comparable with the literature. There was no significant effect of mother's age, nor of the season of previous birth on the length of inter-birth intervals. Inter-birth interval was significantly longer when the first foal was a male. This finding indicates that additional costs of having a son may delay future reproduction and thus reduce the total number of offspring a mare can have during her lifetime. Individually-based data provide critical information on the determinants of reproductive rates, and are therefore a key to understanding the causes of variations in life-history traits.