In order to determine the possibility of using infrared thermography (IRT) in preventing lameness in dairy cows, the aim of this study was to evaluate temperature variations depending on the lesion presence, measurement points and leg position. The study used about 3 000 IRT records from 60 Holstein cows housed in a free-stall barn. Surface temperature measurements were taken at two regions of the hooves: the region of the coronary band, and the region of the skin up to 2 cm above the coronary band. A highly significant ( P = 0.004; 0.006, P < 0.01) difference in temperature was observed between healthy and diseased legs at both measurement points. Additionally, a significant ( P = 0.029; 0.037; 0.045; 0.012; 0.018, P < 0.05) or highly significant ( P = 0.004; 0.006, P < 0.01) difference in temperature values between the front and rear legs was established for both measurement points, i.e., the coronary band and the skin. Despite confirming the potential applicability of infrared thermography in the detection of lameness, it is crucial to consider the significant influences of the environmental factors, impurities, and animal-related factors.