Abstract: The responses of eleven pairs of crib-biting and non-crib-biting horses (controls) to an arousal-inducing stimulus were studied. Video-observation of the horses revealed that crib-biting horses spent between 10.4 and 64.7% of their stabling time performing the stereotypy. During the first 2 days of an experimental period, the horses were conditioned to receive food from a special bucket. On the third day the food bucket was presented, but the horses were not allowed to feed. Arousal behaviour and crib-biting intensity as well as plasma cortisol concentration, heart rate (HR) and heart rate variability (HRV) were recorded at rest, and during and after presentation of the food stimulus. The stimulus induced a significant increase of HR and arousal behaviour in crib-biters and in controls, whereas the crib-biting frequency decreased. Power spectral analysis of the HRV revealed significant differences between crib-biters and controls at rest: crib-biters had a lower vagal tone (high frequency component, HF) and a higher sympathetic tone (low frequency component, LF) than controls. The lower basal parasympathetic activity might be an indication why crib-biting horses, in contrast to the controls, showed neither a significant decrease of the HF component during presentation of the food stimulus nor an increase of the HF component after presentation. Thus, there might be differences in the tuning of the autonomous nervous system and of the stress reactivity in crib-biting and in control horses. The results suggest that the crib-biting horses are more stress sensitive and physiologically and psychologically less flexible than the control horses.