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In this study, the effect of acupuncture on skin temperature (TSKIN), including six body regions (neck, shoulder, ribs, flank, internal thigh, and back), rectal temperature (TRECTAL), serum levels of total proteins, hematocrit, magnesium, calcium (Ca), phosphorus, and chloride was evaluated in five Thoroughbred horses. Horses competed in two official races. For each race, animals were transported from their stables to the racetrack. Horses transported and competed in the first race represent the control group. Two weeks later, the same horses competed in the second race. Before road transport, they were treated with acupuncture. From animals, the TSKIN, TRECTAL, and blood samples were collected at rest; after unloaded, 30 minutes after unloaded, at rest in the transit box, at the end of the race, and 30 minutes after the race. A significant effect of transport and exercise was found on all studied parameters (P < .01), except for Ca and TSKIN of neck (P > .05), in both groups. Transported and exercised horses subjected to acupuncture treatment showed statistically significant higher values of TRECTAL and TSKIN of flank (P < .01). Thoroughbred horses display a classic hemodynamic response to transport and exercise; acupuncture affects body temperature and skin temperature of flank region in both transported and exercised horses suggesting a controlling effect of acupuncture on thermoregulatory function, probably, by decreasing the activities of sympathetic function and manifested as vasodilatation, arterial blood flow, and skin microcirculation increase.Acupuncture treatment on core and surface temperature in the athletic horse.Horses were transported and competed in two official races.Significant effect of transport and exercise was found on studied parameters.Higher values of rectal temperature and skin temperature of flank were found in acupunctured horses.The study suggests a controlling effect of acupuncture on thermoregulatory function.