Provision of nest sites is beneficial for the welfare of laying hens in intensive production systems. The design of these nest sites has a direct effect on pre-laying behaviour. The aim of this study was to investigate the effects of screening off the entrances of the nest boxes with non-transparent flaps and providing nesting or flooring material on pre-laying behaviour. Eighteen individual Hisex brown laying hens and 18 groups of five hens were tested for 2 days in a test cage containing three nest boxes each with a different nesting material (peat, artificial turf and plastic-coated wire mesh). On one of the days the nest boxes were equipped with plastic flaps at the entrances. Pre-laying behaviour was analysed using focal sampling. Both degree of seclusion and nesting material had an effect on pre-laying behaviour of hens. Nest boxes with flaps were visited less frequently but for a longer duration per visit than nest boxes without flaps. Hens spent more time sitting and scratching, and less time standing and moving when flaps were present. Hens also received fewer pecks (group test) when nest boxes were equipped with flaps. Major differences were found between pre-laying behaviour on coated wire mesh compared to peat and artificial turf with shorter duration of nest visits (group test), more standing (group test) and moving (individual and group test), and less sitting (individual and group test) and object pecking (group test) on coated wire mesh. Differences in pre-laying behaviour on peat versus artificial turf were observed for the behaviours moving (less on peat), sitting (more on peat in the individual test) and object pecking (more on peat in the group test). It is concluded that hens show more settled pre-laying behaviour and more nest-building behaviour in the presence of flaps at the entrances of nest boxes. These results indicate that seclusion of nest boxes with non-transparent flaps is beneficial to laying hen welfare. Differences in pre-laying behaviour also suggest that plastic-coated wire mesh is less suitable as nesting material than peat and artificial turf. (C) 2007 Elsevier B.V. All rights reserved.