Background: During the production cycle, dairy animals in intensive production systems are exposed to various challenging events including novel stimuli (e.g., handlers, food and group-mates). The welfare of an individual animal depends on the ability to cope with environmental challenges. Novelty is generally associated with negative emotions however; individual behavioural and physiological responses toward challenging events can be very variable. So, studying behavioural response of calves towards novelty can help us address the welfare needs of the buffalo calves in a holistic manner. Methods: For this study, 24 Murrah buffalo calves were selected at birth and randomly allotted to four groups (six calves each) comprising of SG group (restricted suckling and group housed), SI group (restricted suckling and individually housed), WG group (weaned at birth and group housed) and WI group (weaned at birth and individually housed) for a period of three months. All calves were trained with bottle feeding from day old of age. At 9th week of age, the behavioural response of the calves to novelty was tested in two different novel situations i.e. novel object test and social novelty test using CCTV camera. Result: The results suggest that buffalo calves separated from mother and housed individually showed increased behavioural responses characterized by increased time in decision making, higher standing idle time and more fearful to an unfamiliar calf compared to other groups. It can be concluded that social isolation of the buffalo calves increases fearfulness to novelty hence reducing their welfare in future.