Exploring non-invasive methods to assess pain in sheep

The aim of this study was to determine whether changes in eye temperature, measured using infrared thermography (IRT), and heart rate variability (HRV) can detect moderate levels of pain in sheep. Six ewes received the following treatments: 1) noxious ischaemic stimulus by application of a forelimb tourniquet (S), 2) noxious ischaemic stimulus and flunixin meglumine (S + F), and 3) flunixin meglumine (F). Maximum eye temperature, HRV, mechanical nociceptive threshold, blood pressure and behaviour were recorded for up to 60 min, including 15 min of baseline, 30 min during intervention and 15 min post-intervention. There was a tendency towards a decrease in the heart rate variability parameters RMSSD (the root mean square of successive R-R intervals) and SDNN (the standard deviation of all interbeat intervals) in treatment S compared to treatment F, and a significant increase in the same parameters between test day 1 and 3. A reduction in eye temperature was detected for all treatments during intervention, but no difference was found between S and F and S + N and F during intervention. The eye temperature decreased more in test day 2 and 3 compared to test day 1 during intervention. A significant reduction for both lip licking and vocalisation was observed between test day 1 and 3, and forward facing ears was the ear posture most frequently recorded in test day 1. We suggest that HRV is a sensitive, non-invasive method to assess mild to moderate pain in sheep, whereas IRT is a less sensitive method.