In the mountains, the traditional practice of transhumance is common in dairy production systems to make use of the high-altitude summer pastures. Although the effects of highland grazing have been intensively studied with respect to cow performance and milk and cheese quality, the actual moving of the animals to the highlands and the consequences of this stressor for performance and milk quality in the days immediately following transhumance has not been investigated in detail. The aim of the present study was to compare the effects of a 6-km walk (lasting 1.5 h) to those of a truck transport of 10.5 km (lasting 1 h), simulating cow movement in transhumance systems, as well as a control treatment in which cows were not moved. The experiment included 12 late-lactating Valdostana Red Pied, 12 Montbéliarde, and 12 Holstein cows (i.e., breeds contrasting in genetic merit for milk production). Each cow was subjected to each treatment in a 3-wk Latin square design. Milk yield was measured, and milk and blood samples were taken around the transhumance simulation events. Cows of the 3 breeds responded similarly to both movement treatments. Walking decreased milk yield by 1 kg/milking, but truck transport did not affect milk yield. Both treatments led to an increase in plasma nonesterified fatty acids and milk somatic cell count compared with controls, and truck transport increased milk fat content. Milk coagulation properties were better for Valdostana Red Pied and Montbéliarde cows than for Holstein cows but were not affected by walking or truck transport. Further studies aiming to compare the 3 breeds should include a wider range of response variables over a longer term, including reproduction performance and repeated transhumance.