Abstract: We tested the hypothesis that social defensive responses to the vocalisation of a predator still exist in horses. The recordings of a grey wolf, an Arabian leopard and a golden jackal were played to 20 Konik polski and Arabian mares. Durations of grazing, standing still, standing alert and the number of steps in walk and trot/canter were measured. In one-minute scans, the distances of the focal horse from the reference horse (DIST-RH) and from the nearest loudspeaker (DIST-LS) were approximated. The vocalisation of a leopard aroused the Arabians more than the Koniks (less grazing, stand-still and walk, more stand-alert and trotting/cantering). Koniks showed more relaxed behaviours to the leopard vocalisation (more grazing, stand-still and walk), but high alertness to the wolf playback (stand-alert, trotting/cantering). Spatial formation of the herd of Koniks showed tight grouping (lower DIST-RH) and maintaining distance from the potential threat (DIST-LS) in response to the wolf howling, while the Arabians approached the loudspeakers in linear herd formation when the leopard growls were played. Adult horses responded to potential predation by changing spatial group formations. This ability to apply a social strategy may be one of the explanations for the least number of horses among all hunted farm animal species.

Abstract: Background The increased use of meta-analysis in systematic reviews of healthcare interventions has highlighted several types of bias that can arise during the completion of a randomised controlled trial. Study publication bias has been recognised as a potential threat to the validity of meta-analysis and can make the readily available evidence unreliable for decision making. Until recently, outcome reporting bias has received less attention. Methodology/Principal Findings We review and summarise the evidence from a series of cohort studies that have assessed study publication bias and outcome reporting bias in randomised controlled trials. Sixteen studies were eligible of which only two followed the cohort all the way through from protocol approval to information regarding publication of outcomes. Eleven of the studies investigated study publication bias and five investigated outcome reporting bias. Three studies have found that statistically significant outcomes had a higher odds of being fully reported compared to non-significant outcomes (range of odds ratios: 2.2 to 4.7). In comparing trial publications to protocols, we found that 40-62% of studies had at least one primary outcome that was changed, introduced, or omitted. We decided not to undertake meta-analysis due to the differences between studies. Conclusions Recent work provides direct empirical evidence for the existence of study publication bias and outcome reporting bias. There is strong evidence of an association between significant results and publication; studies that report positive or significant results are more likely to be published and outcomes that are statistically significant have higher odds of being fully reported. Publications have been found to be inconsistent with their protocols. Researchers need to be aware of the problems of both types of bias and efforts should be concentrated on improving the reporting of trials.

Abstract: In recent years, Przewalski’s horses have been increasingly kept in semi-reserves. However,there areonly few studies ontheir behaviour and their ability to adaptto management interventions.In the main part of my dissertation, I focus on investigatingthe animals’ behaviour in different semi-reserves with varyinghabitats and living spaces. In addition, I investigate the horses’ behaviour during various management interventionsand analysetheensuing changes instress levels. Another aspect of my dissertation is the studyof social behaviour inPrzewalski’s horses. I investigate theparameters that should be used to demonstrate social bonds between individualsandassess whichdata provide the most meaningful results.In the commentary tochapter 1,several studies investigatingsocial bonds in horsesare discussed. Comparing the various studies, it is strikingthat no homogeneous analyses orevaluation criteria exist. While some authors only considersocial grooming, others include data onthe spatial proximity of the individuals in their evaluations, and various definitionsof proximity can also be found in the literature. Additionally, someauthors use friendly approaches between individuals asa furtherparameter wheninvestigating the social bonds.Continuing with this theme, in chapter 2I investigate the social behaviour of the horses and compare various analysis methods. I show that proactive behaviour, such as friendly approaches, is a good alternative to spatial proximity when investigating social bonds between group members, andis also useful for expanding the often very small data sets of mutual grooming in horses. Comparing Przewalski’s horses with wild living horses, I found no significant differences in the social behavior and the frequency of social interactions, regardlessof group size, group composition, habitat, and individual parameters such as age and gender.Inchapter 3,I investigate the behaviour of a Przewalski’s horse group when exploring a new area of their enclosure. Their behaviour changed, showing less resting and more feeding. Furthermore, the animals maintained greater distances from each other, and the alpha male, instead of herding the group from behind, led the group around the new area and walked in front of the other group members. Moreover, he showed a substantial increase in stress level during the first day.A general comparison of the behaviour of the Przewalski’s horses in different semi-reserves is provided in chapter 4. In it, the habitat choice of the animals and their reactions to various management interventions are investigated. It is shown that Przewalski’s horses prefer open grassland to dense woods, although keeping Przewalski’s horses in a pine forest does not influence the animals’ stress level. In contrast to habitat, food range, and changes in the group composition, which do not appear to change stress levels, individual factors, such as the hierarchy, influence the glucocorticoid level of the animals significantly. The largest increases in stress hormones were demonstrated when the horses were temporarily confined in smaller areas.The importance of the available space is also discussed in chapter 5, where it is shown that horses show less aggressive behaviour when more space is provided. In contrast, the husbandry system does not influence the animals’ aggression, but the way of feeding can additionally reduce agonistic behaviour, for example if food is offered ad libitum.In summary, the results of this study provide indications for the optimization of keeping Przewalski’s horses in semi-reserves. The animals can adapt themselves to the environment and thrive in habitats which do not correspond to their original steppe-like home. Nevertheless, the semi-reserves should provide sufficient grassland, as the horses prefer this type of habitat. General speaking, any types of habitat can only offer a suitable living space if the food range is sufficient for the number of horses. Otherwise, and especially during could winter months, supplementary feeding is necessary according to the body condition of the animals. This is particularly important for older, weakened, or very young animals, which are still adapting to life in the semi-reserve. Without sufficient food, stress hormones can increase and negatively influence the well-being of the horses. The same is true for management interventions: restricting the animals to small enclosures, for example, can adversely affect the horses’ well-being and should be only done if absolutely necessary. Targetedbehaviour observations allow the animals that have a special meaning for the group to be identified, and these should not be taken out of the group unless it is unavoidable, as young and unexperienced horses orientate themselves on those animals. This is especially true for the alpha male in a bachelor group, as these groups are often composed of young horses and the alpha-male provides the necessary stability and experience. Social bonds between individuals can be investigated by observing friendly and proactive behaviour, and social grooming and friendly approaches yield suitable data for such analysis.

Abstract: The effect of stabling for the first time on the behaviour and welfare of young and naïve horses has not yet been studied in detail. In this study we examined the effect of two typical housing systems on their subsequent behavioural and physiological responses upon first time stabling. Thirty-six 2-year-old Dutch warmbloods, 18 geldings and 18 mares were included in the study. Half of the horses were stabled in individual stables (10.5m2) and the other half in pair housing (48m2 for two horses). The study lasted 12 weeks. At the end of the study the physiological and temperamental responses of the horses on the different treatments was tested using a CRF challenge test (to test the HPA-axis function) and a Novel Object test (to test temperamental differences) respectively. Especially in the first week after stabling pair housed horses spent more time eating whereas individually housed horses spent more time either standing vigilant or sleeping. Stress-related behaviours like neighing, pawing, nibbling and snorting were all displayed significantly more frequently in the individually housed horses (P<0.01). At the end of the study 67% of the individually housed horses was seen performing one or more stereotypies (P<0.01). The cortisol response and ACTH response on the CRF challenge test were lower for horses in the individually housed boxes. It is suggested that this depression in socially isolated animals is caused by a desensitisation of the HPA axis in response to stress-induced elevations in ACTH and cortisol. In general there was no effect of the treatment on the reactivity of the horses during the Novel Object test. However, there were significant relations between the responses of horses in the Novel Object test and in the stable environment. It is concluded that sudden isolated stabling is stressful to young and naïve horses, resulting in a high prevalence of stereotypies and abnormal behaviours. This study also provided some support for the notion that social stress in horses may be associated with a blunted adrenocortical response to CRF challenge. The finding that responses of horses to a behavioural test are correlated with home environment behaviours suggests that individual horses exhibit consistent behavioural traits across different contexts, and opens the possibility of using behavioural tests in horses to predict more general underlying behavioural characteristics.

Abstract: Analysis of heart rate variability (HRV) is a noninvasive approach for investigating the sympathovagal balance of the autonomic nervous system. In recent years, HRV has been increasingly evaluated in animal research. In horses, it has been suggested that basal resting conditions can be achieved by restraining them. The aim of this study was to verify how restriction of movement influences HRV i2n horses. Ten healthy standardbred mares were used to measure the electrocardiographic signal under 2 conditions: free to move in the stall and restrained in the stock. Results indicate that the restriction of movement is associated with increased nervous system sympathetic activity not consistent with resting conditions.