Abstract The aim of this work was to assess the response of Friesian steers to road transportation at low, medium and high ( ca 200, 300 and 600 kg/m 2) stocking densities in terms of physiology, behaviour and carcass bruising. Stocking density was determined by adjusting the number of animals per pen (3 runs) or by adjusting pen size (3 runs). Plasma cortisol and glucose increased ( P < 0·001) with stocking density, as did plasma CK activity ( P < 0·001) and carcass bruising ( P < 0·01) both of which indicate muscle damage. At constant stocking density stress increased with pen location towards the tail of the truck, as reflected by a significant increase in plasma cortisol. Stocking density influenced standing orientation within the truck. At low stocking density animal movement was unrestricted and the preferred orientation was standing parallel to the direction of travel; there was a strong bias against diagonal orientations. Losses of balance were associated with specific driving events, notably braking and cornering. Minor losses of balance were observed at all stocking densities, but major losses of balance including animals going down underfoot were associated with high density. When an animal went down at high stocking density it did so involuntarily and was sometimes trapped down, destabilizing other members of the group in a domino effect. Exploratory, sexual and aggressive behaviours were inhibited at high stocking density, with the exception of mounting and pushing, which increased in frequency with stocking density. It was concluded that the high stocking density adversely affected animal welfare and lowered carcass quality when compared with the medium and low stocking densities.