The defensive burying test is an experimental model that is used to explore anxiety-like behavior in adult rats. Because the expression of anxiety-like behavior may differ between infant and adult rats, we tested the impact of chambers with different sizes and shapes on defensive burying in 28-day-old Wistar rats. The first two chambers had base areas of 560 cm, but one was rectangular and the other round. The base areas of the other two chambers were 282 cm, also with one rectangular and one round. We examined the effects of vehicle and 1 mg/kg diazepam on defensive burying in the various chambers. Locomotor activity was also measured to identify or exclude any sedative effects. Independent of the treatments used, the infant rats showed a shorter burying latency in the three modified chambers and a longer cumulative burying time compared with the original apparatus. The effects of diazepam (i.e. increased latency and decreased burying time) were only significant in the small round chamber, without significant effects on general motor activity. These results suggest that a small round chamber that is used to test burying behavior is sensitive to the anxiolytic actions of diazepam when the experimental subjects are very young rats.