In view of the lack of research on the behaviour of horses working on mechanical treadmills, this study was aimed at assessing the behaviour of ponies on the treadmill and at analysing their heart rate variability in order to describe characteristic types of their behavioural and emotional response. The study was conducted on nine Felin ponies, which were being accustomed to working on a mechanical treadmill for three consecutive days. Their heart rate variability parameters were measured and their behaviour was assessed while the animals were stabled, when they were walking onto the treadmill, during treadmill work and as they walked off the treadmill. The results obtained were used to describe behavioural and emotional response patterns of the horses being accustomed to working on the treadmill. The results were subjected to multivariate analysis of variance (repeated measures), t-Tukey’s test and Spearman’s rank correlations. It was found that the proposed method of describing the types of emotional and behavioural response of ponies indicates the need to assess their level of being accustomed to working on the treadmill by observation of their behaviour combined with simultaneous measurement of their heart rate parameters. The use of both methods is necessary especially when ponies are entering the treadmill and during their work on the treadmill, which are the two most difficult stages for the animals. It is also worth remembering that ponies should first be accustomed to the new environment and only later to working on the treadmill. A three-day period of adaptation to the new environment may be insufficient. Care should be taken especially on the second day of getting ponies accustomed to the treadmill, which can be regarded as critical. A positive finding is that the increased emotional arousal/excitability of ponies trained for treadmill work is short-lived.
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