Metagenomic sequencing reveals high reproducibility of human donor microbiota transplanted into germ-free mice via lower gut route

Yapeng YANG; Xiang TAN; Zeyue ZHANG; Lifeng LIANG; Zhifeng WU; Jinhui HE; Yuqing WANG; Miaomiao DONG; Jixia ZHENG; Hang ZHANG; Shuaifei FENG; Wei CHENG; Bota CUI; Hong WEI; Qinjin LI

doi:10.1631/jzus.B2400495

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Research Article | Updated：2026-04-23

Metagenomic sequencing reveals high reproducibility of human donor microbiota transplanted into germ-free mice via lower gut route
Metagenomic sequencing reveals high reproducibility of human donor microbiota transplanted into germ-free mice via lower gut route
宏基因组测序揭示人源供体菌群经灌肠途径移植至无菌小鼠的高重现性特征
Journal of Zhejiang University-SCIENCE B Vol. 27, Issue 4, Pages: 375-389(2026)
Affiliations：

1.College of Animal Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China
2.State Key Laboratory of Agricultural Microbiology, College of Animal Sciences and Technology, Huazhong Agricultural University, Wuhan 430070, China
3.Central Laboratory, Clinical Medicine Scientific and Technical Innovation Park, Shanghai Tenth People’s Hospital, Tongji University, Shanghai 200072, China
4.Precision Medicine Institute, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou 510080, China
5.Yu-Yue Pathology Scientific Research Center, Chongqing 401329, China
6.Medical Center for Digestive Diseases, The Second Affiliated Hospital of Nanjing Medical University, Nanjing 210003, China
Funds：
DOI：10.1631/jzus.B2400495
CLC：
CSTR：
Received：23 September 2024，

Revised：2025-03-26，

Online First：22 April 2026，

Published：23 April 2026
Accepted：

杨亚鹏,谭向,张泽岳等.宏基因组测序揭示人源供体菌群经灌肠途径移植至无菌小鼠的高重现性特征[J].浙江大学学报（英文版）（B辑：生物医学和生物技术）,2026,27(04):375-389.

Yapeng YANG, Xiang TAN, Zeyue ZHANG, et al. Metagenomic sequencing reveals high reproducibility of human donor microbiota transplanted into germ-free mice via lower gut route[J]. Journal of Zhejiang University-SCIENCE B, 2026, 27(4): 375-389.
杨亚鹏,谭向,张泽岳等.宏基因组测序揭示人源供体菌群经灌肠途径移植至无菌小鼠的高重现性特征[J].浙江大学学报（英文版）（B辑：生物医学和生物技术）,2026,27(04):375-389. DOI： 10.1631/jzus.B2400495.

Yapeng YANG, Xiang TAN, Zeyue ZHANG, et al. Metagenomic sequencing reveals high reproducibility of human donor microbiota transplanted into germ-free mice via lower gut route[J]. Journal of Zhejiang University-SCIENCE B, 2026, 27(4): 375-389. DOI： 10.1631/jzus.B2400495.

摘要

人类菌群相关（HFA）小鼠常被用于模拟人类肠道微生物群结构，以探索疾病与肠道微生物群间的因果关系。然而，人类微生物群在无菌小鼠中的定植效率受多种因素影响，且灌胃和灌肠移植两种途径的定植效果差异尚待阐明。本研究探讨了不同移植途径对受体与供体间肠道微生物的群落结构和功能的重现性的影响，并比较了灌胃（上消化道移植，GT_mice组）和灌肠（下消化道移植，LGT_mice组）两种菌群移植方式在受体小鼠中微生物定植差异。我们通过对每个受体的粪便进行高通量宏基因组测序，并分析各组微生物组组成。结果显示，通过下消化道移植将人类粪便微生物群引入无菌小鼠具有较高的转移效率，这不仅体现在其物种群落结构与供体相似度较高（Adonis分析结果：LGT_mice组-供体组，Adonis

=0.713 960；GT_mice组-供体组，Adonis

=0.774 095），也表现为更高的细菌定植率。上述发现为提高人源化微生物群移植模型的构建准确性提供了新见解，有助于加深对人类肠道微生物群与疾病关联机制的理解。

Abstract

Human flora-associated (HFA) mice are often used to simulate the structure of human intestinal microbiota and to study the causal relationships between diseases and gut microbiota. However

several factors affect th

e colonization efficiency of human microbiota in germ-free (GF) mice

and the differential effects of gavage and lower gut transplantation on colonization are still unclear. In this study

we explored the reproducibility of the recipient-to-donor gut microbiota community structure and function under different transplantation routes and the differences in microbial colonization between recipients via gavage transplantation (GT_mice group) and lower gut transplantation (LGT_mice group). High-throughput sequencing of the metagenome was performed on the feces of each subject

and the composition of microbiome of each group was analyzed. As expected

the introduction of human fecal microbiota into GF mice via lower gut transplantation had a high transfer efficiency

which was evident from the similar species community structure to that of the donor (Adonis

=0.713 960 for LGT_mice group‒donor group; Adonis

=0.774 095 for GT_mice group‒donor group) and a higher bacterial colonization rate. The findings provide unique insights into improving the accuracy of constructing humanized microbiota transplantation models

aiding our understanding of the relationships between the human gut microbiota and disease.

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references

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