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. 2018 Jul 13;8:10646. doi: 10.1038/s41598-018-28764-5

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© The Author(s) 2018

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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Germ-free female bodies are mildly more attractive than conventional female bodies. (A) Inset: A two-way choice courtship assay where conventionally raised (CR) males from fly stocks were aspirated into a chamber containing CV and GF female bodies. Main axes: 2-choice courtship assay of CV vs. GF female over a 60 min period. Only trials displaying courtship ≥ 1 min were analysed. Data was normalized by removing all non-courtship related behaviour throughout the 60 minute period. F1 (n = 92) and F2 (n = 21) courtships were analysed separately. The medians and their confidence intervals are given in green text; the green curves are bootstrap-estimated distributions of the medians. (B) Relative abundance of CHC chemical classes determined by GC/MS. Data represent the average relative abundance (middle line) and confidence intervals (top and bottom lines) of 3 replicate sets per condition, with each set consisting of 8 females. Relative abundance (arbitrary units, a.u.) is calculated by dividing the area under each CHC peak by the area under the hexacosane internal standard peak. Mean differences between CV and GF relative abundance: linear alkane −1.797 (95CI −8.681, +5.087, P = 0.9239), branched alkane +1.651 (95CI −5.232, +8.535, P = 0.9427), diene −1.174 (95CI −8.058, +5.71, P = 0.9830), monoene −1.71 (95CI −8.594, +5.173, P = 0.9355).