## 4 Discussion
In natural environments, factors like varying temperatures, humidity, wind, and uneven terrain can greatly impact thermoregulation and overall physiological performance [45]. For instance, humidity influences evaporative heat loss, while wind speed boosts convective heat loss [46]. Similar findings have been reported by other researchers, who documented an absence of sweating in horses after treadmill exercise under controlled temperature and wind speed conditions [38; 47]. During the ice boot treatment and study period, no edema, cellulitis, skin necrosis, or frostbite was observed due to the cooling method used. Similar observations have been reported by other authors regarding the use of ice boot treatment or cryotherapy [29]. The observed lesions, including dermatitis, cellulitis, alopecia, tissue necrosis, and distal limb edema, resembled frostbite, non-freezing cold injuries, and prolonged water immersion injuries and were reported in patients with multiple health conditions [30]. No signs of distress or incoordination were observed during habituation. The treadmill exercise and thermographic examinations were conducted in the same room under controlled conditions, with a restricted number of participants. The same individuals participated in both procedures. Other authors have suggested that in order to obtain reliable results, it is advisable to minimize the number of individuals present during treadmill exercise testing [18]. Before this treadmill study, there was a period of accommodation to treadmill exercise for 5 days. The horse was habituated with the treadmill by walking and trotting. Other authors have performed the same treadmill habituation process of walking horses for 5 min at a speed of 1.2 m/s and trotting at 2.6-3 m/s [48]. There are different methods that use a cooling temperature of 4 \({}^{\circ}\)C, such as ice water immersion and cold gel wraps, which are applied for 30 min [23; 24; 49]. These methods reduce the cooling time compared to cryotherapy, where the treatment period can be up to 48 h at 5 \({}^{\circ}\)C [24]. In our study, the ice boot pack was maintained on the study limb for 20 min immediately following the training session. At the time of application, the mean temperature of the ice boot was 3.8 \({}^{\circ}\)C.  
For accurate thermal images, a horse's coat and skin must be completely dry and free from sweat and excess hair, and the ambient temperature should remain constant and uniform to avoid external influences on thermographic results [1; 8; 22; 50]. During exercise, the working muscles generate significant amounts of heat, which the body dissipates via sweating to prevent overheating [51]. In our study, the training exercise was performed in an area with a constant temperature of 22 \({}^{\circ}\)C, no air currents, and a humidity level of 70-75%, conditions that did not affect the quality of the thermographic images and did not influence the horse's sweating [47]. The results obtained demonstrated a decrease in the temperatures of the metacarpus and coronary band following treatment with the ice boot pack, which was attributed to vasoconstriction induced by cooling. Further research on cooling methods is required to assess the temperature of the hood wall, both before and after cooling treatment; evaluate the efficacy of cooling methods in horses with conditions such as hoof abscess and laminitis; compare various cooling techniques that facilitate a rapid decrease in temperature and sustain it over extended periods; compare the temperature evolution of the palmar tendons; examine whether the responses are similar between warmblooded, hotblooded, and coldblooded horses; and include variable environmental conditions and field-based study designs to more accurately reflect real-world scenarios. This pilot study presented several limitations that should be considered when interpreting the results. First and foremost, the sample size (\(n=1\)) significantly restricted generalizability, as individual physiological variations may influence thermal responses.