Eye Blink Rates and Eyelid Twitches as a Non-Invasive Measure of Stress in the Domestic Horse

SIMPLE SUMMARY: Eye blink rate has been used as an indicator of stress in humans and, due to its non-invasive nature, could be useful to measure stress in horses. Horses exhibit both full and half blinks as well as eyelid twitches. We exposed 33 horses to stressful situations such as separation from herdmates, denied access to feed and sudden introduction of a novel object, and determined that full and half eye blinks decrease in these situations. Feed restriction was the most stressful for the horse as indicated by increased heart rate, restless behaviour and high head position. The decrease in eye blink rate during feed restriction was paralleled with an increase in eyelid twitches. There was no increase in eyelid twitches or heart rate with the other treatments indicating that the horses did not find these overly stressful, but they did focus their attention more during these situations. Observation of eye blinks and eyelid twitches can provide important information on the stress level of horses with a decrease in eye blinks and an increase in eyelid twitches in stressful environments. ABSTRACT: Physiological changes provide indices of stress responses, however, behavioural measures may be easier to determine. Spontaneous eye blink rate has potential as a non-invasive indicator of stress. Eyelid movements, along with heart rate (HR) and behaviour, from 33 horses were evaluated over four treatments: (1) control—horse in its normal paddock environment; (2) feed restriction—feed was withheld at regular feeding time; (3) separation—horse was removed from visual contact with their paddock mates; and (4) startle test—a ball was suddenly thrown on the ground in front of the horse. HR data was collected every five s throughout each three min test. Eyelid movements and behaviours were retrospectively determined from video recordings. A generalized linear mixed model (GLIMMIX) procedure with Sidak’s multiple comparisons of least squares means demonstrated that both full blinks (16 ± 12(b) vs. 15 ± 15(b) vs. 13 ± 11(b) vs. 26 ± 20(a) full blinks/3 min ± SEM; a,b differ p < 0.006) and half blinks (34 ± 15(ab) vs. 27 ± 14(bc) vs. 25 ± 13(c) vs. 42 ± 22(a) half blinks/3 min ± SEM; a,b,c differ p < 0.0001) decreased during feed restriction, separation and the startle test compared to the control, respectively. Eyelid twitches occurred more frequently in feed restriction (p < 0.0001) along with an increased HR (p < 0.0001). This study demonstrates that spontaneous blink rate decreases while eyelid twitches increase when the horse experiences a stressful situation.