Cull dairy cows are transported to slaughter, but may be more vulnerable to transport stress than younger livestock. In order to ensure the welfare of cull cows during transport their fitness for transport must be assessed before transport. Lameness is a common reason for culling dairy cows, and assessing fitness for transport in lame dairy cows is a frequent task for farmers, veterinarians and livestock drivers. The aim of this study was to evaluate the agreement within and between these three groups of professionals in relation to lameness scoring and assessment of fitness for transport. The study used an online questionnaire consisting of 30 video recordings of walking cows. Participants were asked to score lameness for each cow and assess if the cow was fit for transport or not. Weighted and unweighted kappa were used as a measure of interrater agreement within and between groups. The levels of agreement within and between the three professional groups were at best moderate. Farmers agreed less than moderate within their own group as well as compared to veterinarians and livestock drivers when assessing fitness for transport. In general, it raises concern that the level of agreement on fitness for transport was moderate or even lower. These results call for more focus on the assessment of fitness for transport, including research and possibly training of the different professional groups in order to ensure good animal welfare during transport.