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. 2021 Jun 7;1:21. doi: 10.1038/s43705-021-00021-3

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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 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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Fig. 1 — a Boxplot (median; IQR; range) of bacterial load in foremilk and hindmilk at infant age 5 and 9 months, as well as DNA blank extraction controls. Statistical significance was evaluated by paired t-tests. Asterisks indicate **p < 0.01 and ***p < 0.001. b, c Scatter dot plots of bacterial load in foremilk versus hindmilk at infant age 5 (a) and 9 (b) months. A linear regression line was fitted to the log-transformed data and the relationship was evaluated by Spearman’s rank correlation analyses. d–f Boxplots (Median; IQR; Range) of bacterial alpha diversity measures, Shannon index (d), Observed ASVs (e) and Evenness index (f), comparing foremilk and hindmilk at both infant age 5 and 9 months. Statistical significance was evaluated by paired t-tests and asterisks indicate *p < 0.05. g, h PCoA plots based on Bray–Curtis (g) and Jaccard (h) dissimilarities, including foremilk and hindmilk samples at both infant age 5 and 9 months, with statistically differential clustering evaluated by ANOSIM tests. i, j Violin plots of the absolute abundances of the major genera (average relative abundance cutoff across all milk samples of 1.0%) comparing hindmilk to foremilk at infant age 5 months (i) and 9 months (j). Thick black lines indicate median and thin black lines 25 and 75 percentiles. Statistical difference was evaluated by Wilcoxon-signed rank tests and symbols indicate ^#FDRp = 0.05–0.1, *FDRp < 0.05, and **FDRp < 0.01.