For wild animals, being in captivity in wildlife centers can cause considerable stress. Therefore, it is necessary to establish and validate non-invasive tools to measure chronic stress during rehabilitation. Eight Common Buzzards which lived in permanent husbandry were placed individually into prepared aviaries and their feces were collected before, during and after a stress event for biological validation over a period of seven days. The extracted fecal glucocorticoid metabolites (fGCMs) were analyzed with three different enzyme immune assays (EIA) to find the most suitable one. Additionally, we aimed to investigate the stability of fGCM levels after defecation because further metabolization by bacterial enzymes can lead to changed results. The Cortisone-EIA performed best in males and females and showed that the stress event led to an fGCM increase of 629% (557% in females and 702% in males) in relation to basal values. We found no significant differences between the sexes, but observed significant differences between different times of day. FGCM concentration significantly changed after eight hours at room temperature. Our study successfully validated the non-invasive measurement of fGCM as a stress indicator in Common Buzzards and could therefore lay the foundation for future studies providing new insights for animal welfare research in Buzzards.

The poultry industry is an important and still growing sector in many parts of the world. For ethical reasons and due to increased consumer awareness for animal welfare in production animals, it is of importance to establish a reliable and objective test system for monitoring and improving health and welfare. During the rearing process, broiler chickens are exposed to numerous potential stressors and management interventions (e.g. weighing of individual animals, preslaughter fasting and capture processes), but assessing the level of stress perceived by the animals entirely through behavioral observations can be challenging. Monitoring stress-related physiological markers, such as glucocorticoids, can be an accurate and presumably more objective addition. To avoid additional stressors induced by blood collection, a noninvasive approach using urofecal samples is advisable. However, a thorough validation is needed to establish a suitable test system for measuring stress hormone levels, including potential effects of the time of day of collection or the time that has elapsed since defecation. Therefore, the aim of this study was to test the stability of urofecal glucocorticoid metabolites (ufGCM) postdefecation, to determine time of day effects on ufGCM levels, and to investigate the effect of standard management procedures on ufGCM concentrations in broiler chickens. Our results revealed a time window of 4 h in which fecal samples from broilers can be collected without major alterations to the ufGCM concentrations. In this regard, a "fecal box" proved useful for collecting uncontaminated fresh samples. The time of day of sample collection did not influence ufGCM concentrations significantly. Moreover, the used assay proved to be sensitive enough to detect even small and short-lasting activations of the HPA axis induced by handling, confinement, and fasting. Thus, the system used can be a powerful and easy to apply tool in a chicken production setup for assessing stress as a marker of welfare in commercially housed broiler chickens, which in the long-term can also improve production, particularly with regard to process quality.

Foot-and-mouth disease (FMD) is a transboundary animal disease that has a major impact on livestock production, regional and international trade and livelihoods of smallholder farmers in endemic settings. Livestock movement is one of the most important ways of spreading infectious diseases including FMD. Many livestock diseases are transmitted through direct contact between animals, and thus between herds and flocks through animal movements. In this study, we described the pattern of livestock movements among smallholder farmers within a communal farming area in South Africa. A cross-sectional survey using a semi-structured questionnaire was administered to 116 respondents, and 13 focus group discussions employing participatory mapping and semi-structured interviews were conducted among smallholder goat farmers. Data from the study reported 37 nodes and 78 ties with an overall network density of 0.059 (SD 0.235) for goats, and 42 nodes and 90 ties with an overall network density of 0.052 (SD 0.223) for cattle across the study area. The study identified several FMD high-risk locations to prioritise vaccination programmes and targeted disease surveillance. Four locations within the (former) FMD-free zone of the country were identified to have connections with movement of goats from the study area. Findings from this study further demonstrated that goats are moved without official movement permits to the FMD free zone of the country, with most farmers being ignorant of the need to obtain official veterinary movement permits. These animal movements put the country at risk of future FMD outbreaks within the free zone. We recommend that the relevant authorities implement risk-based control measures to prevent the spread of infectious diseases.

In South Africa, blue wildebeest (Connochaetes taurinus) are routinely captured for relocation purposes. To monitor the stress caused by this practise, a non-invasive method assessing adrenocortical function as a measure of stress would minimize disturbance during sample collection. In our study, an adrenocorticotropic hormone challenge, and a mass-capture event were used to examine the suitability of five enzyme immunoassays (EIAs) for monitoring stress-related physiological responses using faeces as a sample matrix. The tested 11-oxoaetiocholanolone I EIA performed best, showing a 2126% increase above baseline after 22 h for a male, and a 474% increase for a female 23 h post-ACTH injection. Baseline faecal glucocorticoid metabolite (fGCM) concentrations did not differ between animals captured on either day 1 or day 2 of the capture event, indicating that the frequent presence of a helicopter during the two days did not influence fGCM concentrations. However, during capture-related restraint for up to 10 h, an overall 1.5-fold elevation in (fGCM) concentrations was found. Storage of faeces at ambient temperature post-defecation indicated a fair stability of fGCMs for up to 8 h. The ability to reliably assess adrenocortical function provides a solid basis to examine endocrine responses to putative stressful circumstances in blue wildebeest.

Little information exists on the sexual behaviours of the roan antelope (Hippotragus equinus; É. 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.

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 welfare. This study examined the impact of human interactions and ride-based activities on physiological stress-related indicators in African elephants. Fifteen trained semi-captive elephants, as well as free-ranging elephants roaming under the same ecological conditions, were monitored. Faecal samples were collected over a nine-month period from both groups and these were analysed using an enzyme immunoassay detecting faecal glucocorticoid metabolites (fGCMs) with a 5β-α-ol-11-one structure. Elephants that participated in elephant-back-safari (EBS) activities showed significant decreases in fGCM concentrations when EBS were discontinued. Similarly, fGCM concentrations of the trained semi-captive individuals that did not participate in EBS showed decreased steroid concentrations over the same time. Overall, fGCM concentrations of the trained semi-captive herd and the free-ranging herd did not differ significantly. The collected data will help to better understand the physiological and behavioural requirements of semi-captive elephants with frequent exposure to humans. The findings will also help to optimise management strategies for wild elephant populations and elephants living in controlled environments on reserves exposed to wildlife tourism.

Infectious diseases and parasitic infestations can cause a set of non-specific clinical signs, such as increased body temperature and resting, and a decrease in food intake. These physiological and behavioural changes have an adaptive function facilitating defences against the pathogen and to support immune functions. These so-called' sickness behaviours' can also be used as an early detection tool for disease. Foot-and-mouth disease (FMD) still causes great economic losses in endemic countries, especially to smallholder farmers. The aim of this study was to determine if behavioural changes in goats can be used as an early indicator of FMD virus (FMDV) infection. The efficacy of a Southern African Territories (SAT) FMD vaccine was studied on forty South African indigenous goats. Changes in daily activities (resting, feeding, walking), as well as social behaviours (social resting, social feeding, dominance behaviours) were recorded and then compared over time and between clinically affected and unaffected goats. Pedometers were used to estimate average daily steps and to compare between groups of study animals. Eleven goats developed clinical signs of FMD, as well as non-FMD related sicknesses during the course of the study. Overall walking and resting behaviours were not significantly affected by the presence of FMD related clinical signs (p > 0.05). However, during the time of FMDV infection, social resting increased significantly (p < 0.001). Although goats developed FMD lesions on lips and tongues, percentage of time feeding was not affected (p = 0.762), suggesting that the study goats did not perceive the oral lesions as an important disturbance. Similarly, the number of steps did not consistently decrease in the presence of FMD-associated foot lesions. When affected by non-FMD related sicknesses, animals did not have an overall reduction in the time spent feeding (p = 0.867). However, goats affected with non-FMD conditions reduced the amount of social feeding (p = 0.002), potentially avoiding energetically costly competition at the feeding points. Overall, goats affected with FMD did not show more sickness behaviour, suggesting that FMDV infection in goats might not lead to obvious and therefore, easily detectable behavioural changes. This might have implications for farmers and animal health personnel, as individual goats infected with FMDV might be undetected within a flock due to the absence of obvious sickness behaviours, and the virus can therefore be spread more easily between herds through animal movements.

Intensive pig management involves in a commercial setting the housing and implementation of certain procedures, such as castration and tail docking, which may be stressful for the animal. Good farming practices include the reduction of stress due to management processes, but assessing the level of stress perceived entirely through behavioural observations can be challenging. The monitoring stress-related physiological markers, like glucocorticoids (GC), can be an accurate alternative that would presumably be more objective. In order to avoid an additional stressor by taking blood, a non-invasive approach is advisable. We used an adrenocorticotropic hormone (ACTH) stimulation test and the effect of transport to examine the suitability of different enzyme immunoassays (EIAs) for monitoring adrenocortical function in domestic pigs using saliva and faeces as sample matrices. An assay measuring faecal glucocorticoid metabolites (fGCMs) with a 3ß,11ß-diol group has proven suited to determine adrenocortical activity, showing an overall increase of 180% in fGCM concentrations related to ACTH administration and of 70% related to transport, respectively. A cortisol EIA was used to detect salivary glucocorticoid (sGC) concentrations, revealing a 1100% increase in sGC concentrations after ACTH administration. The stability of fGCM concentrations post-defecation was determined to assess possible changes in measured fGCM concentrations in unpreserved faecal material over time, with fGCM concentrations being relatively stable (maximal 12% change) under natural conditions for approximately two days after defecation. This implicates that untreated faecal material from pigs can be analysed for up to two days after collection without appreciable level of depreciation in fGCM concentrations. Being able to assess the physiological stress response of domestic pigs non-invasively can help to improve the well-being of commercially reared pigs.

ContentsAs the natural habitat of more and more species becomes depleted, captive breeding programmes have become established to bring species back from the brink of extinction. Monitoring the reproductive status of an individual is essential in order to improve breeding success. Traditional methods have involved stressful blood sampling, and thus noninvasive methods have been proven to be reliable alternatives for monitoring reproductive function in both captive and free‐ranging animals. Subsequently, noninvasive methods have become an invaluable tool in longitudinal studies and conservation efforts, as animals can be observed without, or minimal human contact. The Beira antelope is a small antelope endemic to the northern part of the Horn of Africa. Population numbers of the Beira have been declining over the last few decades due to habitat fragmentation. We show here that the reproductive cycle of female Beira antelopes can be monitored noninvasively, by using faecal samples to analyse oestrogen (fEM) as well as progestagen (fPM) metabolites. The profiles of fPM and fEM of both females showed regular cyclic patterns in which the follicular and luteal phases could be distinguished. The overall mean cycle length is 22 days (range: 21–25 days), with a mean length of the follicular phase of 6 days (range: 4–7 days) and a mean length for the luteal phase being 15 days (range: 12–16 days). The suitability of these noninvasive techniques should assist in optimizing breeding efforts of this endemic small antelope in captivity. Being noninvasive, this method could also be a useful tool for monitoring reproductive function in the dwindling wild populations.

Fission-fusion social societies allow animals to respond in a flexible manner to environmental changes by adapting the size and composition of a group. Although group members change frequently in these systems, associations with preferred partners may be found. In this study, we examined the grouping patterns of a population of 80 individual giraffes in a fenced South African game reserve over a 12-month period. Using social network analyses as a tool to evaluate observed associations, we subsequently analysed both sex- and season-related grouping patterns within the study population. Mixed sex groups represented 49% of all groups observed, and although overall group composition distribution did not differ significantly between seasons, the number of encountered single females decreased by 50%, whereas the number of multi-male groups increased by over 50% in winter. Overall average group size did not differ significantly between seasons, but significantly larger multi-female and multi-male groups were seen in winter. Within the social network, two distinct clusters were found in summer, with the population more divided in winter, with five distinct clusters emerging. The strongest ties (highest HWIGs) were found between adult females. Our study revealed that giraffes not only live in a highly flexible social fission-fusion system, but also show seasonal patterns of grouping.

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.

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. (C) 2017 Elsevier Inc. All rights reserved.

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.

Poster presented at the University of Pretoria, Faculty of Veterinary Science Faculty Day, August 25, 2016, Pretoria, South Africa.

Measuring energetic condition of wild animals is of major importance in ecological research, as it is profoundly linked to fitness. However, noninvasive monitoring of energetic condition in wild-living animals is methodologically challenging. Measuring urinary C-peptide levels is a suitable method to noninvasively assess energy balance in wild-living animals. As collecting urine is not always feasible in the wild, it is essential to establish alternative biomarkers for other sample types to assess energy balance. Thyroid hormones (TH) are potential candidates as they are involved in the regulation of metabolic processes. During periods of low energy intake, serum TH levels are reduced, leading to a decrease in metabolic activity. To investigate whether fecal TH can serve as a biomarker for energy balance, we validated a total T-3 ELISA to measure immunoreactive T-3 (iT(3)) in fecal samples of yellow-breasted capuchins. We restricted caloric intake of seven males, assessed daily group caloric intake and determined daily individual fecal iT(3) levels. Analytical validation of the assay showed that fecal iT(3) levels can be reliably measured; however, proper storage conditions must be implemented and possible degradation to be accounted for. IT3 levels were significantly higher on days with high group caloric intake. However, individual iT(3) levels varied substantially, resulting in an overlap across individuals between conditions. Our results indicate that fecal iT(3) levels can serve as a useful biomarker to detect changes in energy intake of yellow-breasted capuchins. Overall, measuring fecal iT(3) levels may present a suitable method for monitoring energy balance when urine collection is impossible.