Abstract: Glucocorticoids (GCs; i.e. cortisol/corticosterone) are a central component of the stress response and thus their measurement is frequently used to evaluate the impact of stressful situations. Their metabolites from faeces of various animal species are more and more taken as a non-invasive aid to assess GC release and thus adrenocortical activity. The current literature review includes an extensive collection (1327 papers) and evaluation (see also Supplementary Tables) of the literature on faecal cortisol/corticosterone metabolite (FCM) analysis published to date. It aims at giving reference for researchers interested in implementing FCM analysis into their study or seeking to improve such methods by providing background knowledge on GC metabolism and excretion, conveying insights into methodological issues and stating caveats of FCM analysis and by highlighting prerequisites for and some examples of a successful application of such methods. Collecting faecal samples and analysing FCMs may appear simple and straightforward, but researchers have to select and apply methods correctly. They also need to be aware of the many pitfalls and potentially confounding factors and, last but not least, have to carefully interpret results. Applied properly, measurement of FCMs is a powerful non-invasive tool in a variety of research areas, such as (stress) biology, ethology, ecology, animal conservation and welfare, but also biomedicine.

Abstract: Tbourida is a traditional Moroccan equestrian sport in which 15 horses gallop 200 m in a line while riders fire into the sky with muskets. The stop is the finale and representative demands of this equestrian event. Such particular sudden stop after a fast gallop requires a hyperextension of the metacarpophalangeal joint. Indeed, it is well known that Tbourida show predisposes horses to different injuries of the hard and soft tissues of the distal forelimbs. Yet, there is a paucity of research that examined such lesions. The aim of the present study was to investigate for the first time the type and the prevalence of osteoarticular findings in the distal forelimbs of Tbourida horses using radiographic images. The study was conducted on 127 Tbourida horses aged between 2.5 and 15 years old with 6-year-old horses being the most affected. Data analysis showed that 93,7% of horses exhibit degenerative joint lesions of the fetlock, 86,6% showed ossification of the ungual cartilage, 78,7% had enthesophytes associated with the deep digital flexor tendons, 81,1% had enthesophytes associated with the suspensory ligament branches, and 19,6% showed a particular exostosis on the first phalanx. This large number of lesions reflects how this sport is difficult for horses and also argues that animals are suffering from a lack of welfare and care in their husbandry management.

Abstract: Abstract Across the globe, wildlife populations and their behaviours are negatively impacted by people. Protected areas are believed to be an antidote to increasing human pressures but even they are not immune to the impact of anthropogenic activities. Areas that have been set aside for the protection of wildlife therefore warrant more attention when investigating the impact of anthropogenic pressures on wildlife. We use cheetahs Acinonyx jubatus as a case study to explore how a large carnivore responds to anthropogenic pressures inside wildlife areas. Using GPS-collar data we investigate cheetah space use, both when moving and stationary, and movement parameters (speed and turn angles) in relation to human disturbance, distance to human settlement, livestock abundance and livestock site use inside wildlife areas. Space use was negatively influenced by human disturbance, resulting in habitat loss and fragmentation and potentially reducing landscape permeability between neighbouring wildlife areas. Cheetahs were also less likely to stop in areas where livestock numbers were high, but more likely to stop in areas that were frequently used by livestock. The latter could reflect that cheetahs are attracted to livestock however, cheetahs in the study area rarely predated on livestock. It is therefore more likely that areas that are frequently used by livestock attract wild herbivores, which in turn could influence cheetah space use. We did not find any effects of people and livestock on cheetahs? speed and turn angles which might be related to the resolution of the data. We found that cheetahs are sensitive to human pressures and we believe that they could be an indicator species for other large carnivores facing similar challenges. We suggest that further research is needed to determine the levels of anthropogenic pressures needed to maintain ecological integrity, especially inside wildlife areas.

Abstract: Identifying valid indicators to assess animals' emotional states is a critical objective of animal welfare science. In horses, eye wrinkles above the eyeball have been shown to be affected by pain and other emotional states. From other species we know that individual characteristics, e.g. age in humans, affect facial wrinkles, but it has not yet been investigated whether eye wrinkle expression in horses is systematically affected by such characteristics. Therefore, the aim of this study was to assess how age, sex, breed type, body condition and coat colour affect the expression and/or the assessment of eye wrinkles in horses. To this end, we adapted the eye wrinkle assessment scale from Hintze et al. (2016) and assessed eye wrinkle expression in pictures taken from the left and the right eye of 181 horses in a presumably neutral situation, using five outcome measures: a qualitative first impression reflecting how worried the horse is perceived by humans, the extent to which the brow is raised, the number of wrinkles, their markedness and the angle between a line through both corners of the eye and the topmost wrinkle. All measures could be assessed highly reliable with respect to intra- and inter-observer agreement. Breed type affected the width of the angle (F2, 114 = 8.20, p < 0.001), with thoroughbreds having the narrowest angle (M = 23.80, SD = 1.60), followed by warmbloods (M = 28.00, SD = 0.60), and coldbloods (M = 31.00, SD = 0.90). None of the other characteristics affected any of the outcome measures, and eye wrinkle expression did not differ between the left and the right eye area (all p-values > 0.05). In conclusion, horses' eye wrinkle expression and its assessment in neutral situations was not systematically affected by the investigated characteristics, except for 'breed type', which accounted for some variation in 'angle'; how much eye wrinkle expression is affected by emotion or perhaps mood needs further investigation and validation.

Abstract: Additional measures of well-being would be beneficial to the management of a variety of species in human care, including elephants. Immunoglobulin A (IgA) is an immune protein associated with pathogen defense, which has been demonstrated to decrease during times of stress, and increase in response to positive stimuli. This paper describes the development and validation of an enzyme immunoassay (EIA) for the quantification of Asian elephant (Elephas maximus) IgA in feces, saliva, urine, and serum. Samples were collected weekly from four females for 6 months to assess IgA and glucocorticoid (GC) concentrations, establish relationships between these two biomarkers, and determine variability in IgA within and between individuals, and across sample types. IgA was quantified in all four sample types, although urinary concentrations were low and sometimes undetectable in individual samples. Concentrations were highly variable within and between individuals, with fecal, salivary and serum IgA, and fecal, salivary and urinary GCs all differing significantly across individuals. Contrary to previous findings, IgA and GC were generally not correlated. Serum IgA was less variable within individuals, with the exception of one female that experienced a brief illness during the study. However, marked inter-individual differences were still apparent. When data from all individuals were combined, fecal IgA was significantly predicted by salivary and urinary IgA; however, this relationship did not hold when individuals were analyzed separately. Analysis of a fifth female that exhibited a more severe systemic illness demonstrated clear increases in fecal IgA and GC, suggesting these may also be useful health biomarkers. Further investigation is needed to determine what sample type is most reflective of biological state in elephants, and how IgA concentrations are associated with health and positive and negative welfare states. Based on observed variability, a longitudinal approach likely will be necessary to use IgA as a measure of well-being.