Abstract: Abstract Horses (Equus caballus) have a rich history in their relationship with humans. Across different cultures and eras they have been utilized for work, show, cultural rituals, consumption, therapy, and companionship and continue to serve in many of these roles today. As one of the most commonly trained domestic animals, understanding how horses learn and how their relationship with humans and other horses impacts their ability to learn has implications for horse welfare, training, husbandry and management. Given that unlike dogs and cats, domesticated horses have evolved from prey animals, the horse-human relationship poses interesting and unique scientific questions of theoretical value. There is still much to be learned about the cognition and behaviour of horses from a scientific perspective. This review explores current research within three related areas of horse cognition: human-horse interactions, social learning and independent learning in horses. Research on these topics is summarized and suggestions for future research are provided.

Abstract: Abstract Only few quantitative reports exist about the concentrations and induction of immunoglobulin A (IgA) in mucosal secretions of horses. Despite this, it is widely assumed that IgA is the predominant immunoglobulin on mucosal surfaces in the horse. Here, two new monoclonal antibodies (mAbs) against equine IgA, clones 84-1 and 161-1, were developed and characterized in detail. Both IgA mAbs specifically bound monomeric and dimeric equine IgA in different applications, such as Western blots and fluorescent bead-based assays. Cross-reactivity with other equine immunoglobulin isotypes was not observed. The new IgA mAb 84-1 was used in combination with the previously characterized anti-equine IgA mAb BVS2 for the development and validation of a fluorescent bead-based assay to quantify total IgA in equine serum and various secretions. The IgA assay's linear detection ranged from 64 pg/ml to 1000 ng/ml. For the quantification of IgA in serum or in secretions an IgA standard was purified from serum or nasal wash fluid (secretory IgA), respectively. The different standards were needed for accurate IgA quantification in the respective samples taking the different signal intensities of monomeric and dimeric IgA on the florescent bead-based assay into account. IgA was quantified by the bead-based assay established here in different equine samples of healthy adult individuals. In serum the median total IgA was 0.45 mg/ml for Thoroughbred horses (TB, n = 10) and 1.16 mg/ml in Icelandic horses (ICH, n = 12). In nasopharyngeal secretions of TB (n = 7) 0.13 mg/ml median total IgA was measured, and 0.25 mg/ml for ICH (n = 12). Saliva of ICH (n = 6) contained a median of 0.15 mg/ml, colostrum of Warmbloods (n = 8) a median of 1.89 mg/ml IgA. Compared to IgG1 and IgG4/7 quantified in the same samples, IgA appeared as the major immunoglobulin isotype in nasopharyngeal secretions and saliva while it is a minor isotype in serum and colostrum. The newly developed monoclonal antibodies against equine IgA and the resulting bead-based assay for quantification of total IgA can notably improve the evaluation of mucosal immunity in horses.

Abstract: Abstract Stress is a generic concept describing the body's reaction to external stimuli, including both physiological and psychological factors. Therefore, by definition, the assessment of psychological stress in the exercising horse encompasses the problem of teasing apart the psychological and physiological factors both of which result in stress responses. The present study reviews the existing literature on various measures of stress taken specifically in the context of equitation science. Particular attention has been paid to short-term effects, and commonly used measurements of short-term stress include heart rate, a number of heart rate variability parameters, blood or saliva cortisol levels, eye temperature, and various behaviour parameters including in particular behaviour patterns presumably indicative of conflict with the rider's/trainer's aids. Inspection of the individual studies' results revealed that disagreement between these different measures of stress is commonplace. For physiological parameters, the largest proportion of agreement (i.e. both parameters simultaneously indicated either higher, insignificant or lower stress compared to a control treatment) was found for heart rate and heart rate variability parameters, while generally limited agreement was found for cortisol. It appears that cortisol levels may not be particularly useful for assessing/assessment of the valence of a situation in the exercising horse as cortisol levels are predominantly linked to activation and exercise levels. Although heart rate variability parameters reflect in theory more closely sympathovagal balance compared to cortisol levels, great care has to be taken regarding the use of appropriate time-frames, appropriate raw data correction methods as well as the use of appropriate equipment. In spite of its wide-spread and apparently successful use, popular equipment may in fact not be accurate enough under field conditions. Eye temperature is another promising parameter for assessment of psychological stress, but the technique is likewise susceptible to application errors. Given the high susceptibility of physiological parameters to errors at various experimental stages, behavioural rather than physiological parameters may in fact provide more accurate measures of valence when conducting experiments in the exercising horse. Behavioural parameters that appear to be particularly practical in assessing stress in ridden horses' behaviour are associated with frequencies of behaviour indicative of conflict. However, while increased frequencies of are a good indicator of stress, the absence of conflict behaviour does not provide proof of the absence of stress due to the possible occurrence of conditions such as Learned Helplessness. In future studies, the above issues should be taken into consideration when designing experiments to assess psychological stress in ridden horses.

Abstract: Abstract There is increasing interest in keeping horses in groups, but progress is hampered by a lack of knowledge about which horses can and should be kept together. Therefore, our objective was to investigate the effect of group composition on the occurrence of injuries among horses, the ease of removing horses from groups and horses’ reactivity to a fearful stimulus. Using a matched case control design, 61 groups of horses were studied in Denmark, Norway, Finland and Sweden. They were allocated into groups of similar or different age and sex or where membership changed regularly or remained stable. Injuries were recorded before mixing the horses into treatment groups, the day after mixing and four weeks later. Reactivity of horses to a moving novel object and the behaviour of a horse being removed from its group and the reactions of other group members towards this horse and the handler were evaluated. It was hypothesized that a more socially variable group composition has beneficial effects on behaviour, ease of handling and reducing reactivity whereas frequent changes in group composition has negative consequences, resulting in more injuries. We found that differences in treatment effects were mainly related to breed, rather than group composition. Icelandic horses reacted less to the movement of the novel object (P = 0.007) and approached it more afterwards (P = 0.04). They also had fewer new injuries than warmbloods following mixing (P < 0.001) and fewer than all other groups 4 weeks later (P < 0.01). Most new injuries after mixing were minor and recorded on the horse’s head, chest, hind legs and rump. In conclusion, variations in sex and age composition of the group had little effect on injury level, reactivity and ease of handling compared to the general effect of breed. Concerns about the risk of severe injuries associated with keeping horses in groups are probably overestimated. Thus, we propose that horses can be successfully kept in groups of different sex and age composition.

Abstract: Abstract— We present a study focused on a quantitative estimation of a human-horse dynamic interaction. A set of measures based on magnitude and phase coupling between heartbeat dynamics of both humans and horses in three different conditions is reported: no interaction, visual/olfactory interaction and grooming. Specifically, Magnitude Squared Coherence (MSC), Mean Phase Coherence (MPC) and Dynamic Time Warping (DTW) have been used as estimators of the amount of coupling between human and horse through the analysis of their heart rate variability (HRV) time series in a group of eleven human subjects, and one horse. The rationale behind this study is that the interaction of two complex biological systems go towards a coupling process whose dynamical evolution is modulated by the kind and time duration of the interaction itself. We achieved a congruent and consistent
statistical significant difference for all of the three indices. Moreover, a Nearest Mean Classifier was able to recognize the three classes of interaction with an accuracy greater than 70%. Although preliminary, these encouraging results allow a discrimination of three distinct phases in a real human-animal interaction opening to the characterization of the empirically proven relationship between human and horse.