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. 2021 Jan 18;11:1727. doi: 10.1038/s41598-020-80826-9

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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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Phylogenetic tree analysis based on distance matrix calculated by multiple sequence alignment (MSA). (a) Neighbor-joining phylogenetic tree analysis of the Bifidobacterium strains identified in both methods. The 16S reference sequences were obtained from the SILVA 138v non-redundant ribosomal RNA gene database. The pink, orange, and green boxes indicate the clustered ASVs by Bifidobacterium species. (b) MSA with the reference sequence for the Bifidobacterium strains and ASVs identified from V3V4 and sFL16S. The alignment analysis was performed by the IUB multiple alignment matrix with options (transition weight 0.50; delay-divergent cutoff 30%) using MEGA X software. The MSA results visualized using the NCBI Multiple Sequence Alignment Viewer 1.16.1v.