The aim of this research paper was to evaluate the effect of a slow-release milk replacer on health and behaviour of neonatal dairy calves. This was done with the potential benefits to welfare during transport in mind. A total of 15 calves were randomly divided into three groups of 5, namely, a control group fed twice in 24 h with 3 l of a conventional milk replacer, a slow-release group fed once in 24 h with 2 l of conventional milk replacer and 1 litre of a specialised micro-encapsulated feed and an enriched-replacer group fed once in 24 h with 3 l of milk replacer enriched with micellar casein. Blood samples were taken before feeding and 6, 12, 18 and 24 h after and analysed for acid–base parameters, electrolytes, glucose, haemoglobin, cortisol, insulin, cholecystokinin and adiponectin. Calf behaviour was recorded between 6 and 14 h after feeding. There was a significant increase in blood pH 6 h after feeding in all groups, but the glucose, HCO3− and base excess increased significantly in the slow-release group only, whereas sodium increased significantly in the enriched group only. Glucose levels remained significantly higher in the slow-release group, relative to the control, at 6, 12, and 18 h after feeding. Insulin levels changed significantly over time in the enriched and control group but remained constant in the slow-release group. Insulin levels were significantly higher in the control group when compared to the slow-release group after feeding. Adiponectin changed significantly over time after feeding in the control group only, but no significant changes were observed between the feeding groups. Behavioural patterns were similar in control and slow release groups but less favourable (less lying time, more vocalisations) in the enriched group. In conclusion, once-daily feeding of slow-release milk replacer demonstrated favourable patterns of blood variables related to satiety and hunger as well as behavioural patterns that did not differ from conventional twice-daily feeding.