Abstract: The aim of the study was to investigate whether heart rate variability (HRV) could assess alterations of the autonomic nervous system (ANS) at different levels of excitement. The behavioural and physiological responses of 20 warmblood horses to a challenging ground exercise task were studied. Prior to the experiment, the horses were evaluated at rest and during forward walking (FW). The horses were then forced to move backwards continuously during 3 min according to a standardised protocol (BW1). Subsequently, the horses were exposed to two training sessions, after which the backward walking (BW2) was re-evaluated. Heart rate (HR) and HRV-parameters such as the standard deviation of the beat-to-beat intervals (SDRR), the low (LF; sympathetic tone) and high frequency (HF) component of HRV (HF; parasympathetic tone) and their ratio (LF/HF; index representing the sympatho-vagal balance) were sampled at rest, and during FW, BW1 and BW2. Stress-related behaviour during BW1 and BW2 was determined from video recordings. The results of the different evaluations were compared to each other. Compared to rest and FW, the first backward experiment induced a significant rise in HR, LF and LF/HF and a significant decrease of HF. SDRR decreased from both FW and rest with only the latter reaching significance. In BW2 after the training sessions, HR and the parameters of the sympathetic branch of the ANS (LF, LF/HF) were decreased and the vagal tone (HF) increased compared to BW1; all changes were significant. The duration of stress indicating behavioural patterns revealed also a significant decrease of excitement after the training, when backward walking did not differ from forward walking in any parameter. Correlations between HRV-parameters and stress indicating behaviour as well as HR were found. We conclude that the HRV-parameters LF and HF are valuable measures to quantify sympatho-vagal balance, which allows a more precise assessment of the responses of HR and SDRR to mental stress during low intensity exercise.

Abstract: The benefits to humans of equine-assisted therapy (EAT) have been well-researched, however few studies have analyzed the effects on the horse. Understanding how differing mental states of humans affect the behaviour and response of the horse can assist in providing optimal outcomes for both horse and human. Four humans clinically diagnosed and under care of a psychotherapist for Post-Traumatic Stress Disorder (PTSD) were matched physically to four neurotypical control humans and individually subjected to each of 17 therapy horses loose in a round pen. A professional acting coach instructed the control humans in replicating the physical movements of their paired PTSD individual. Both horses and humans were equipped with a heart rate (HR) monitor recording HR every 5secs. Saliva samples were collected from each horse 30 min before and 30 min after each trial to analyze cortisol concentrations. Each trial consisted of 5 min of baseline observation of the horse alone in the round pen after which the human entered the round pen for 2 min, followed by an additional 5 min of the horse alone. Behavioural observations indicative of stress in the horse (gait, head height, ear orientation, body orientation, distance from the human, latency of approach to the human, vocalizations, and chewing) were retrospectively collected from video recordings of each trial and analyzed using a repeated measures GLIMMIX with Tukey's multiple comparisons for differences between treatments and time periods. Horses moved slower (p < 0.0001), carried their head lower (p < 0.0001), vocalized less (p < 0.0001), and chewed less (p < 0.0001) when any human was present with them in the round pen. Horse HR increased in the presence of the PTSD humans, even after the PTSD human left the pen (p < 0.0001). Since two of the PTSD/control human pairs were experienced with horses and two were not, a post-hoc analysis showed that horses approached quicker (p < 0.016) and stood closer (p < 0.0082) to humans who were experienced with horses. Horse HR was lower when with inexperienced humans (p < 0.0001) whereas inexperienced human HR was higher (p < 0.0001). Horse salivary cortisol did not differ between exposure to PTSD and control humans (p > 0.32). Overall, behavioural and physiological responses of horses to humans are more pronounced based on human experience with horses than whether the human is diagnosed with a mental disorder. This may be a reflection of a directness of movement associated with humans who are experienced with horses that makes the horse more attentive. It appears that horses respond more to physical cues from the human rather than emotional cues. This knowledge is important in tailoring therapy programs and justifying horse responses when interacting with a patient in a therapy setting.

Abstract: Responses of horses towards fear-eliciting stimuli can have important consequences for both human and horse safety. This experiment was designed to investigate behavioural and heart rate (HR) responses of horses to novel visual, auditory and olfactory stimuli. Twenty-four 2-year-old, previously unhandled, stallions were habituated to receive a food reward from a container in a test arena. Each horse was exposed to three 2 min tests in a balanced design where in addition to the feed container, either a traffic cone (visual test), white noise (auditory test) or eucalyptus oil applied to the inside of the container (olfactory test) were used as the novel stimuli. Compared to the control, less time was spent eating during all tests. There was no difference in locomotion activity in the different test situations, but presentation of the novel visual and auditory stimuli elicited significantly increased HR responses in the horses, compared to their response to the arena without novel stimuli (control), whereas there was no increase in HR response to the olfactory stimulus. However, during the olfactory test, the horses had an increased number of eating bouts and became more vigilant towards their surroundings, whereas during the visual and auditory tests, more time was spent alert towards the stimulus. The horses also took significantly more steps backwards in response to the auditory test. The heart rate responses correlated between tests and reflect a non-differentiated activation of the sympathetic nervous system, while the behavioural responses were linked to the type of stimulus.

Abstract: Recent interest has focused on the advantage of intensively handling young foals as a means of producing more tractable horses, accustomed to humans and receptive to training. To date, the effect of this intensive handling, dubbed “imprint training” in the popular literature, has not been tested. The present study compares seven foals handled intensively as neonates with eight untreated controls. The handling protocol started from 2-8 h after birth and continued daily for a total of 5 days. The protocol consisted of a series of stimuli and experiences that were each repeated until the foal no longer resisted or reacted negatively. Subsequently, foals were tested before weaning, at 4 months of age. Results indicated that handled foals (HF) ranked higher than control foals (CF) in subjective ratings of calmness (*P<0.0117) and friendliness (*P<0.0001) and in several specific handling tasks (venipuncture *P<0.0220; placing in stock *P<0.0128). Although, in approach tests all foals but one allowed approach of a person to 4 m, significantly more HF approached the person than CF (P<0.0080). In stimulus tests, foals were presented specific stimuli to which they had been tested as neonates. Two of eight CF were too unruly and dangerous to test. Of foals that could be tested, CF required significantly more time to hook-up a heart rate monitor (**P<0.0055). Split-plot analysis indicated that HF had lower heart rates to initial left-sided stimuli, presented first, than CF (*P<0.0421). In response to right-sided stimuli, heart rate scores of CF were not significantly different from HF (P<0.2259), suggesting reduced reactivity over time due to a learning effect. Behavioral responses to specific stimuli did not differ between CF and HF, suggesting that neonatal handling has a general rather than specific effect on subsequent behavior. Cortisol concentrations were measured before and after testing and the difference calculated. All foals had higher post-testing levels than pre-testing levels. There was a significant difference between HF and CF, indicating greater reactivity among the CF (*P<0.050). In general, the results indicated that foals handled as neonates were more tractable and less reactive. Specific neonatal handling tasks, such as sticking a finger up the foal's nose or patting the bottom of the foot, seemed to have no beneficial effect on related tasks such as passing a nasogastric tube or tapping with a farrier's hammer at 4 months of age. Mechanisms for the observed effect of neonatal handling require further investigation.

Abstract: Horses frequently react nervously when passing animal production farms and other places with distinctive smells, leading riders to believe that horses are innately frightened by certain odours. In three experiments, we investigated how horses respond to (1) urine from wolves and lions, (2) blood from slaughtered conspecifics and fur-derived wolf odour, and (3) a sudden auditory stimulus in either presence or absence of fur-derived wolf odour. The experiments were carried out under standardised conditions using a total of 45 naive, 2-year-old horses. In the first two experiments we found that horses showed significant changes in behaviour (Experiments 1 and 2: increased sniffing; Experiment 2 only: increased vigilance, decreased eating, and more behavioural shifts), but no increase in heart rate compared to controls when exposed to predator odours and conspecific blood in a known test environment. However, the third experiment showed that exposure to a combination of wolf odour and a sudden stimulus (sound of a moving plastic bag) caused significantly increased heart rate responses and a tendency to a longer latency to resume feeding, compared to control horses exposed to the sudden stimulus without the wolf odour. The results indicate that predator odour per se does not frighten horses but it may cause an increased level of vigilance. The presence of predator odour may, however, cause an increased heart rate response if horses are presented to an additional fear-eliciting stimulus. This strategy may be adaptive in the wild where equids share habitats with their predators, and have to trade-off time and energy spent on anti-predation responses against time allocated to essential non-defensive activities.