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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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Pre-exposure to ethanol does not change aversive and appetitive olfactory learning. Before the larvae were conditioned, they were separated from their food and treated for 20 min with either water or 20% ethanol. They were then washed and trained with the odors AM and BA and either 1.5 M NaCl or 0.01 M fructose as a teaching signal. (a) Larvae treated with water and 20% ethanol both showed an aversive odor-salt memory (one-sample t test, pwater < 0.001, ppre-EtOH < 0.001) that was not different from each other (paired t-test, p < 0.112). Sample size for each box plot is n = 15. (b) Larvae treated with water and 20% ethanol both showed an appetitive odor-fructose memory (one-sample t test, pwater < 0.001, ppre-EtOH < 0.001) that was not different from each other (paired t-test, p < 0.484). Sample size for each box plot is n = 15. 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.