Temporary maternal separation is a stressful situation typical under modern husbandry practices and may have an undesirable impact on lamb welfare. However, at present there is insufficient evidence on temporary maternal separation effects on the behaviour of juvenile lambs. The aim of the present study was to assess if and how partial isolation during different periods at the early age can induce differences in the expression of juvenile lamb behaviour at the age of 2 and 3 months old. Twenty-seven male Chios lambs were assigned to 3 experimental groups: C (control), S1 (lamb daily isolation for 1h between the 13th and 20th day of age) and S2 (lamb daily isolation for 1h between the 20th and 27th day of age).At the age of 2 and 3 months old, the behaviour of lambs was tested by the implementation of 2 types of test: isolation and novel object (Y-test). There were no statistical differences between lamb treatments in the number of jumps, vocalizations and heart rates during isolation test and the latency to approach a novel or a familiar object in the novel object test conducted at 2 and 3 months of age. However, lambs showed a tendency of turning to the right-hand side of the test pen, irrespective of treatment group, at the age of 2 and 3 months old during the implementation of novel object test.It is concluded that no discernible effects of maternal temporary deprivation on the display of stress induced behavioural components of juvenile lambs at the age of 2 and 3 months old were found.