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. 2022 Dec 16;12:21768. doi: 10.1038/s41598-022-26335-3

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© The Author(s) 2022

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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Mucus collection procedure. (a) Commonly available kitchen sieves with decreasing mesh size (X = 1 × 1 mm, Y = 0.75 × 0,75 mm, Z = tightly woven filaments) were used to filter the stiff and insoluble fraction of the mucus. (b) Fraction of the mucus left after filtration with X; Y = (c); Z = (d). (e) Detail of the Z’s mesh filaments under microscope (however, any sieve with mesh size of 0.5 × 0.5 mm down to 0.2 × 0.2 mm could be used as effectively). (f,g) After incubation of the slugs in the PBS buffer the slugs spontaneously crawl out of the containers to be collected, rinsed gently with tap water, and left to recover.