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. 2021 Jun 10;11:12307. doi: 10.1038/s41598-021-91677-3

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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The effects of ethanol on larval olfactory attraction. (a) Scheme of the experimental procedure. The olfactory stimuli 1-OCT, AM, BA, and 8% ethanol were presented on a 2.5% agarose plate in custom made Teflon containers to prevent direct contact of the animals with the chemical. Larvae were allowed to crawl for 5 min. (b) Canton-S larvae preferred all odors including 8% ethanol over empty containers (n^1-Octanol = 19, Pref^1-Octanol = 0.17, ci = 0.04–0.29, n^AM = 8, Pref^AM = 0.51, ci = 0.3–0.69, n^8%Ethanol = 16, Pref^8%Ethanol = 0.33, ci = 0.24–0.41, n^BA = 16, Pref^BA = 0.65, ci = 0.55–0.72). All groups were significantly different to zero (one-sample t test, p^1-octanol < 0.011, p^AM < 0.001, p^8%Ethanol < 0.001, p^BA < 0.001). (c) Scheme of the experimental procedure. Test plates contained 2.5% agarose mixed with either 5%, 8%, 10%, or 20% ethanol. BA was presented in Teflon containers. Larvae were allowed to crawl for 5 min. (d) Canton-S larvae showed an olfactory preference for BA irrespective of the ethanol concentrations in the test plate. All groups were significantly different to zero (one-sample t test, p^BA-5%Ethanol < 0.001, p^BA-8%Ethanol < 0.001, p^{BA-10%Ethanol} < 0.001, p^{BA-20%Ethanol} < 0.001), but not different from each other (ANOVA, p < 0.697). Sample size for each box plot is n = 16. Differences against zero are indicated in red at the bottom of each panel. Significant differences of two groups are specifically indicated with an asterisk. Non-significant results are not indicated. Preference scores and statistical tests underlying the different indices are documented in the Supplementary material.