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This study aimed to validate eye temperature (ET) assessed using infrared thermography (IRT) as an indicator of welfare in horses. Moreover, this study aimed to determine the most accurate position for ET measured using IRT and to validate this approach as a gold-standard measurement method. As the quantitative data obtained by IRT have strongly influenced the ET results depending on the specific location of the measurement area, an accurate definition of the regions of interest (ROI) was established. A total of 176 horses (Thoroughbred, Warmblood, and Halla horses) were used at the racing course of the Korea Racing Authority and public horse-riding clubs in South Korea. The present study also compared temperatures among three ROIs of the eye—lacrimal sac, medial canthus, and lateral canthus—at rest. Correlations between ET, rectal temperature (RT), heart rate (HR), and respiratory rate (RR) were assessed. There were no significant correlations between HR, RR or RT; however, among the three ROIs, the temperature of the medial canthus was positively correlated with RT (p &lt; 0.05). Furthermore, the size of ROI was negatively correlated with accuracy of temperature measurement. These results indicate that the most suitable area for ET measurement using IRT in horses is the medial canthus, and it is recommended to use the average temperature of the smallest ROIs (2 × 2 pixels) for temperature analysis. Therefore, this study offers a validated protocol in which ET measured using IRT in the horses is useful as an indicator of welfare.

Validation of Eye Temperature Assessed Using Infrared Thermography as an Indicator of Welfare in Horses