The aim of this study was to evaluate the effects of age of separation on the behavioural responses of the dairy calf and cow. Calves were separated from their dams 6 h, 1 day, or 4 days after birth (n=9 cow-calf pairs in each of the 3 treatment groups) and behaviour was video and audio taped from 1 h before separation to 21 h after separation. In the hour immediately before separation, we found that the younger calves tended to call and move more in the pen, and spent more time standing than the older calves, but after separation these trends reversed. Calves separated at older ages made significantly more movements in the pen (P<0.05), spent more time standing (P<0.05) and spent more time with the head out of the pen (P<0.01) than calves separated soon after birth. We observed a similar pattern for the cows. Before separation, cows with younger calves moved more frequently about the pen (P<0.05), and called at much higher rates (a mean of 40.7 calls during 40 min for cows on the 6-h treatment, vs. 0.2 calls for cows in the 4-day group; P<0.001). After separation, cows in the 4-day group called at approximately four times the rate of those separated at 6 h or 1 day (P<0.01). Moreover, the calls produced by cows separated later had a significantly higher fundamental frequency (P<0.001) and a lower emphasized harmonic (P<0.02) than the calls of cows separated from calves soon after birth. There was no difference between treatment groups in the other behavioural measures, either before or after separation. Calves separated at older ages tended to require fewer days of treatment for scouring, but calf weight gain and cow milk production did not differ among treatment groups. In conclusion, behavioural responses of both the cow and calf increase in relation to calf age at separation. However, there may be health advantages associated with delayed separation that compensate for the increased behavioural response.