ABATRACT We investigated 10 suckling cross-bred calves (Japanese Black (Wagyu) x Holstein) and collected data on the physiological stress responses of the young calves to long-haul road transportation. All calves were male and 21-47 days of age. The calves were obtained at a livestock market and were transported from the market to the university in a commercial livestock vehicle. The total haul distance and time were 306.9 km and 7 h, respectively. The calves were deprived of food and water during transportation. Blood samples, electrocardiograms, and rectal temperatures were collected immediately after transportation and after one week of habituation at the university. Serum pH was significantly lower (P < 0.01), while serum triiodothyronine, aspartate aminotransferase, non-esterified fatty acids (all P < 0.05), and heart rate (P < 0.01) were significantly higher immediately after transportation. The concentrations of blood lactic acid (P = 0.08) and serum alanine aminotransferase (P = 0.06) tended to be higher after transportation. These physiological responses suggest that the stress caused by long-haul transportation causes significant effects on liver function in young calves.