1Horses are routinely travelled for access to safe off-road riding, veterinary care, 2 breeding, sale or moving to a new home environment. However, transport is a known 3 stressor in horses. For this reason, problem behaviour when loading is a commonly 4 reported issue which presents risks to handlers and horse welfare. Existing literature 5 and manufacturers recommendations suggests that alpha-casozepine may be 6 effective in improving the behaviour and welfare of horses during loading onto a 7 vehicle for transport. The current paper aims to assess the behavioural and 8 physiological effects of a commercially available alpha-casozepine feed supplement 9 (Zylkéne Equine) in horses during loading and confinement on a transport lorry. 10 Subjects (n = 10) were loaded once with the supplement and once without, in a 11 balanced random order with each subject acted as their own control. The handler 12 was blind to treatment. Time to load onto the lorry, and movement of feet, licking and 13 chewing, and vocalising within the lorry, were recorded as behavioural indicators of 14 compliance and coping. Heart rate, heart rate variability, salivary cortisol, and 15 infrared thermography of both core temperature and the discrepancy between eyes, 16were measured as indicators of arousal. There were no significant differences in 17 physiology between Treatment and Control (P > 0.05). Treatment resulted in a 18 significantly shorter Loading Time than control (P = 0.04), however, the actual 19 difference in median time was only 0.45 seconds. No other behavioural indicator 20 differed between Treatment and Control (P > 0.05). Power analysis revealed the 21 sample was sufficient to detect a significant effect. Where modest effects were 22 observed for a small number of variables, Treatment effect contradicted predictions. 23Taken together, this indicates that alpha-casozepine does not affect a horse's ability 24 to cope with loading and confinement in a horse lorry. Further work is required to ascertain whether the maximum dosagetwice that used heremight affect coping 26 and behaviour in horses. 27 28