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. 2017 Aug 23;7:9231. doi: 10.1038/s41598-017-09112-5

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Perspective view of the double-roofed flight tunnel. The floor, roof and left wall of the flight tunnel consisted of planks with red and white stripes oriented transversely to the bees’ flight paths. One side of the tunnel consisted of insect netting lined with stripes formed by a red gelatin filter. The 25 cm-wide and 220 cm-long tunnel consisted of two parts: (i) the first half had a permanent 54-cm low roof, and (ii) the second half had a double roof consisting of a removable inner part set at a height of 54 cm and a permanent upper part set at a height of 1 m. (A) Under constant low roof conditions, the inner roof was set at the height (54 cm): the tunnel is therefore endowed with an uniform height of 54 cm. (B) Under “dorsal ditch” conditions, this inner roof was removed to make the 1-m high upper roof visible to the honeybees, thus imposing a sudden perturbation on their dorsal visual stimuli. Honeybees were trained to enter the tunnel under constant low roof conditions near either the roof or the floor, depending on the experiment, and they were able to collect a sugar solution reward placed in a box at the end of the tunnel either near the roof or near the floor, accordingly.