Inter-breed diversity and temporal dynamics of the faecal microbiota in healthy horses.

Understanding gut microbiota similarities and differences across breeds in horses has the potential to advance approaches aimed at personalized microbial modifications, particularly those involved in improving sport athletic performance. Here, we explore whether faecal microbiota composition based on faecal 16S ribosomal RNA gene sequencing varies across six different sport breeds at two time points 8 months apart within a cohort of 189 healthy horses cared for under similar conditions. Lusitano horses presented the smallest and Hanoverians the greatest bacterial diversity. We found subtle but significant differences in ß-diversity between Lusitano, Anglo Arabian and the central European breeds, and we reproduced these results across the two time points. Repeat sampling of subjects showed community to be temporally more stable in Lusitano and Anglo Arabian breeds. Additionally, we found that 27 genera significantly varied in abundance across breeds. Overall, 33% of these taxa overlapped with previously identified taxa that were associated with genetic variation in humans or other species. However, a non-significant correlation was observed between microbial composition and the host pedigree-based kinship. Despite a notable variation in the diversity and composition of the faecal microbiota, breed exerted limited effects on the equine faecal microbiota.

Stereotypic horses (Equus caballus) are not cognitively impaired.

Stereotypies in animals are thought to arise from an interaction between genetic predisposition and sub-optimal housing conditions. In domestic horses, a well-studied stereotypy is crib-biting, an abnormal behaviour that appears to help individuals to cope with stressful situations. One prominent hypothesis states that animals affected by stereotypies are cognitively less flexible compared to healthy controls, due to sensitization of a specific brain area, the basal ganglia. The aim of this study was to test this hypothesis in crib-biting and healthy controls, using a cognitive task, reversal learning, which has been used as a diagnostic for basal ganglia dysfunction. The procedure consisted of exposing subjects to four learning tasks; first and second acquisition, and their reversals. For each task, we measured the number of trials to reach criterion and heart rate and heart-rate variability. Importantly, we did not try to prevent crib-biters from executing their stereotypic behaviour. We found that the first reversal learning task required the largest number of trials, confirming its challenging nature. Interestingly, the second reversal learning task required significantly fewer trials to reach criterion, suggesting generalisation learning. However, we did not find any performance differences across groups; both stereotypic and control animals required a similar numbers of trials and did not differ in their physiological responses. Our results thus challenge the widely held belief that crib-biting horses, and stereotypic animals more generally, are cognitively impaired. We conclude that cognitive underperformance may occur in stereotypic horses if they are prevented from crib-biting to cope with experienced stress.

Gut microbiota resilience in horse athletes following holidays out to pasture.

Elite horse athletes that live in individual boxes and train and compete for hours experience long-term physical and mental stress that compromises animal welfare and alters the gut microbiota. We therefore assessed if a temporary period out to pasture with conspecifics could improve animal welfare and in turn, favorably affect intestinal microbiota composition. A total of 27 athletes were monitored before and after a period of 1.5 months out to pasture, and their fecal microbiota and behavior profiles were compared to those of 18 horses kept in individual boxes. The overall diversity and microbiota composition of pasture and control individuals were temporally similar, suggesting resilience to environmental challenges. However, pasture exposure induced an increase in Ruminococcus and Coprococcus that lasted 1-month after the return to individual boxes, which may have promoted beneficial effects on health and welfare. Associations between the gut microbiota composition and behavior indicating poor welfare were established. Furthermore, withdrawn behavior was associated with the relative abundances of Lachnospiraceae AC2044 group and Clostridiales family XIII. Both accommodate a large part of butyrate-producing bacterial genera. While we cannot infer causality within this study, arguably, these findings suggest that management practices maintained over a longer period of time may moderate the behavior link to the gut ecosystem beyond its resilience potential.

Horses Could Perceive Riding Differently Depending on the Way They Express Poor Welfare in the Stable.

This study investigated the relationships between four behavioral and postural indicators of a compromised welfare state in loose boxes (stereotypies, aggressive behaviors toward humans, withdrawn posture reflecting unresponsiveness to the environment, and alert posture indicating hypervigilance) and the way horses perceived riding. This perception was inferred using a survey completed by the usual riding instructor and during a standardized riding session (assessment of behaviors and postures, qualitative behavior assessment (QBA) and characterization of the horses' locomotion using an inertial measurement unit). In accordance with ear and tail positions and the QBA, stereotypic and the most hypervigilant horses in loose boxes seemed to experience a more negative affective state during the riding session compared with nonstereotypic and less hypervigilant animals (P < .02 in all cases). Horses which were aggressive toward humans in loose boxes had higher scores regarding the occurrence of discomfort and defensive behaviors on the survey than nonaggressive horses (P = .03). They also presented higher dorsoventral accelerations at a canter during the riding session (P = .03), requiring the rider to increase his spinal movement (P = .005). These results suggest that aggressive horses may be harder to ride than nonaggressive animals. The expression of unresponsiveness to the environment in loose boxes was related to more reluctance to move forward, as assessed in the survey (P = .006). This study suggests that a compromised welfare state in the stable is related to horses having a more negative perception of riding. This perception could vary depending on the expression of poor welfare.

Priming for welfare: gut microbiota is associated with equitation conditions and behavior in horse athletes.

We simultaneously measured the fecal microbiota and multiple environmental and host-related variables in a cohort of 185 healthy horses reared in similar conditions during a period of eight months. The pattern of rare bacteria varied from host to host and was largely different between two time points. Among a suite of variables examined, equitation factors were highly associated with the gut microbiota variability, evoking a relationship between gut microbiota and high levels of physical and mental stressors. Behavioral indicators that pointed toward a compromised welfare state (e.g. stereotypies, hypervigilance and aggressiveness) were also associated with the gut microbiota, reinforcing the notion for the existence of the microbiota-gut-brain axis. These observations were consistent with the microbiability of behaviour traits (> 15%), illustrating the importance of gut microbial composition to animal behaviour. As more elite athletes suffer from stress, targeting the microbiota offers a new opportunity to investigate the bidirectional interactions within the brain gut microbiota axis.

Housing Horses in Individual Boxes Is a Challenge with Regard to Welfare.

Horses are mainly housed in individual boxes. This housing system is reported to be highly detrimental with regard to welfare and could trigger the expression of four behavioural indicators of a compromised welfare state: stereotypies, aggressiveness toward humans, unresponsiveness to the environment, and stress-related behaviours. The aim of this study was to identify housing and management factors that could alleviate the detrimental effects of individual boxes on welfare. A total of 187 horses were observed over 50 days by scan sampling. The impact of 12 factors was investigated on the expression of the four behavioural indicators in three different analyses. The results show that the majority of factors tested did not influence the expression of the behavioural indicators. Only three (straw bedding, a window opening onto the external environment, and a reduced quantity of concentrated feed) would have beneficial, although limited, effects. Furthermore, the longer the horses spent in individual boxes, the more likely they were to express unresponsiveness to the environment. To preserve the welfare of horses, it seems necessary to allow free exercise, interactions with conspecifics, and fibre consumption as often as possible, to ensure the satisfaction of the species' behavioural and physiological needs.

Progressive habituation to separation alleviates the negative effects of weaning in the mother and foal.

Early and definitive separation between offspring and their mothers has negative consequences on behavioral and physiological responses. This study compared sudden and definitive weaning (Sudd group, N = 16) and weaning involving progressive habituation to separation using a fence line during the month preceding definitive separation (Prog group, N = 18). The impact of these two methods was assessed in both foals and their mothers through behavioral and biological parameters, including salivary cortisol, telomere length and blood transcriptomes. On the day of definitive separation, Prog foals neighed and trotted less and presented lower cortisol levels than Sudd foals. The weaning type also acted on the foals' personality development; Prog foals became more curious, less fearful and less gregarious than Sudd foals, and the effects remained visible for at least 3 months. In principal component analysis, the Sudd and Prog groups were well separated along a factor where fear, reactivity and gregariousness correlated with high cortisol levels, but curiosity was associated with an increased telomere length and higher expression of genes involved in mitochondrial functions. Progressive weaning was also beneficial in mares. Principal component analysis showed that most Sudd group mares had higher cortisol levels and displayed more alert postures, neighs and activity on the day of weaning, indicating higher stress levels, while Prog mares had profiles that were characterized by more time spent resting on the day of weaning and longer telomere lengths. In conclusion, this study shows that progressive habituation to separation alleviates the negative effect of definitive weaning on both the mother and her young compared to sudden separation.