Current procedures for ranching and sustainable use of guanacos necessitate their transport. Transportation is a risky process for animals, and is a particular concern for wild-caught or semi-domesticated species such as the guanaco — a wild South American camelid species increasingly being established on farms in Chile and Argentina. This study investigated the effect of transport on the physiological and behavioural responses of eight castrated adult male guanacos, transported on a single 2 h journey at a stocking density of 113.5 kg m-2 (0.76 m2 per animal). Plasma cortisol and blood glucose concentration, total and differential white blood cell (WBC) counts, heart rate, and body weight were measured one week before, immediately before, immediately after, 2 h after and one week after transport. Behavioural responses were recorded during handling prior to loading. Immediately after transport we found significant increases in plasma cortisol concentrations and neutrophil:lymphocyte (N:L) ratio, the latter peaking 2 h after transport. Heart rate increased significantly only during loading, while body weight remained constant throughout. Behavioural responses related to handling (jumping, vocalising, kicking, spitting and urinating) were not associated with the physiological response. All variables returned to pre-transport values within one week. Transport of guanacos under these conditions produced physiological changes similar to those associated with a mild and transient stress response in other species and which, we judge, fall comfortably within acceptable limits for their welfare.