Piling behaviour of laying hens often results in smothering or death due to suffocation. Mechanisms leading to piling are not yet understood though various potential factors have been suggested. In this experimental study, we predicted that the presence of a light spot, a novel object (metal foil), or a heat area within animal pens would increase animal numbers around the stimulus leading to piling behaviour. We presented the cues in a 4 × 2 Latin-square design in eight identical experimental pens including each 55 Lohmann Selected Leghorn hens. The cues were presented in two test areas per pen, at two bouts per day in the morning, consecutively for 5 days, over four periods (age: 20, 22, 24, 26 weeks). Each pen received a cue and control condition simultaneously (test areas without cue presentation) once. For a bout, each cue was presented for 35 min except for the light spot where the duration was 10 min. Birds’ responses to the cues during bout and non-bout times were video recorded and analysed for the first bout of each period. To assess the cues’ attractiveness, the number of hens during bout times was counted at predefined times within the test and control areas. To assess the cues’ effects on piling, we described piling behaviour (pile number, duration, animal numbers, trigger) in control and test areas during bout times. Furthermore, we described piling behaviour during bout times and non-bout times on the first day of the first period and fourth period. The best model explaining the number of hens included the interactions of treatment and bout time, and treatment and area. Over the bout’s time course, more hens were attracted to the light spot compared to the control condition, and more to test areas compared to control areas. In the novel object condition, more hens were drawn to the test areas compared to the control areas. Hens were not attracted to the heat area. Piling in bout times was observed twice when hens pecked at the novel object. During non-bout times, piling behaviour occurred frequently at midday and in the late morning compared to the afternoon, mostly in corners and mainly preceded by the mutual attraction of hens. Overall, hens were attracted to light spots and less so to the novel object though neither reliably induced piling behaviour. The occurrence of piling behaviour in non-bout times shows that more work is needed to understand mechanisms eliciting piling behaviour.
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