2Tongue-ties (TT) are commonly applied to both Standardbred and Thoroughbred racehorses to increase control, 3 by preventing them from getting their tongue over the bit, and as a conservative treatment for equine respiratory 4 conditions, principally dorsal displacement of the soft palate. This study investigated responses to TT application 5 in horses, at rest, using both behavioural (head-tossing, ear position, gaping and lip licking) and physiological 6 (salivary cortisol concentrations, eye surface temperature and heart rate) indices. Twelve Standardbred horses (six 7 of which were naïve to TT) were used in a randomised crossover design. The study comprised 3 phases; Phase 1 8 (Baseline), Phase 2 (Treatment), and Phase 3 (Recovery). At phase 2, tongue tie application (TTA) was performed 9 using a rubber band that was looped around the tongue and secured to the mandible for 20 minutes. The control 10 treatment (C) incorporated 30 secs of tongue manipulation, at the start of the 20 min, however no TT was applied.
11Behaviours (head-tossing, ear position, mouth gaping and lip-licking) and heart rate (HR) were recorded for the 12 duration of the study and analysed in ten minute intervals. Salivary samples were taken at the end of each phase 13 for subsequent cortisol assays and infrared thermography images were taken of each eye at 5-minute intervals.14 Statistical analyses were performed in SPSS using linear mixed effects models and repeated measures general 15 linear models, to determine differences between treatments and within treatments, over time. Compared to control,
16there was more head-tossing/shaking (p<0.001), gaping (p<0.001) and backwards ear position (p<0.001) and less 17 forward ear position (p<0.001) during TTA, in Phase 2. Horses with previous experience of TT showed more 18 head-tossing (p=0.040) and gaping (p=0.030) than naïve horses. Lip-licking was more frequent after TTA 19 treatment than control, during Phase 3 (p<0.001). Salivary cortisol concentrations increased after TTA 20 (1846.1pg/mL ± 478.3pg/mL vs 1253.6pg/mL ± 491.6pg/mL, p=0.047). Mean HR, and mean right and left eye 21 temperature did not differ significantly between treatments in any phase (all p> 0.05). The findings of this study 22 suggest the application of a tongue-tie causes changes to both behavioural and physiological parameters suggestive 23 of a stress-related response. Further research is needed that will enable racing and sport horse regulatory bodies 24 to make informed decisions about the appropriate use of tongue-ties in horses. 25 26