One aspect of farm animal adaptability is behavioural and physiological responses toward challenging situations, often implying elements of novelty. Dairy cows show strong behavioural responses when presented with novel food and the behavioural responses reflect the degree of novelty in the food and have an acceptable repeatability. In the present study two experiments were conducted to examine whether the behavioural responses toward novel food were affected by short-term food deprivation or whether the metabolic load in terms of increased milking frequency and decreased energy density in the food affected behavioural and adrenocortical responses of dairy cows toward novel food. The effects of metabolic load and 12 h of food deprivation on the behavioural and physiological responses toward novel food were examined in dairy cows provided with 4 kg of carrots as novel food. In Experiment 1, effects of 12 h of food deprivation were tested using dairy cows provided with 4 kg of carrots as novel food after 0 h versus 12 h of food deprivation. Food deprivation did not affect the approach-avoidance behaviour toward novel food but led to increased attempts to steal usual food from neighbours as well as decreased self grooming. In experiment 2, effects of milking frequency and energy density in the food on behavioural and physiological responses toward novel food were examined. The cows were milked either twice or three times daily. The energy in the food after calving was low or high. An extra daily milking affected the approach-avoidance behaviour of the cows as shown by reduced time spend away from the novel food and reduced frequency of sniffing the novel food. Low energy density in the food led to increased time spent away, increased latency to eat novel food, decreased food intake and a higher proportion of cows performing self-grooming. Provision of novel food did not affect the plasma concentration of cortisol sampled after food provision, and no differences were found between treatments. In conclusion, 12 h of food deprivation did not affect behavioural responses toward novel food in dairy cows, suggesting that this test of behavioural responses toward novelty is rather insensitive toward day-to-day changes in eating motivation. Low energy density in the daily ration and increased milking frequency affected the behavioural, but not the adrenocortical responses toward novelty but there was no evidence for a simple relationship between the energy balance of dairy cows and their responses toward novel food.