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Effects of Different Preparation Methods on Microbiota Composition of Fecal Suspension

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Abstract

Fecal microbiota transplantation is an emerging disease-modifying therapy. The viability of the microbiome in feces and its successful transfer depends on the preparation of fecal microbiota suspension. However, currently, no standard operation procedure is proposed for fecal suspension preparation. This study aims to compare the effect of different preparation methods on the composition of fecal microbiota composition in the rat. Four methods were used to collect the fecal suspension from fresh rat fecal (Group A), including stirring with normal saline (Group B), stirring with normal saline and then standing (Group C), stirring with normal saline and filtered with gauze (Group D), and stirring with normal saline and centrifuged (Group E). 16S ribosomal RNA gene (16S rDNA) sequencing technology was used to analyze the microbiota diversity and composition of each group of samples. Compared with fresh feces, the bacterial richness of the fecal suspension obtained by the four methods was significantly decreased (P < 0.05). The structural similarity with fresh fecal microbiota from high to low is groups B, D, C, and E. All four methods changed the microbiota structure to varying degrees, thus may affect the effect of FMT. In conclusion, choosing different methods to prepare fecal suspensions may help to better optimize the application of FMT.

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Data Availability

The data generated in the study have been uploaded to the NCBI Sequence Read Archive (SRA) under accession PRJNA827478. Now the data are available for reviewers (https://dataview.ncbi.nlm.nih.gov/object/PRJNA827478?reviewer=lqau50mvmdm0jilpgasc99ln3r).

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Funding

This research was funded by the Zhejiang Provincial Natural Science Foundation of China (Nos. LGF19H030006 and LQ20H030001), Ningbo Science and Technology Project (No. 2019C50100), and Ningbo Clinical Medicine Research Center Project (No. 2019A21003).

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Authors and Affiliations

  1. The Affiliated Lihuili Hospital, Ningbo University, Ningbo, 315046, Zhejiang, People’s Republic of China

    Fangxia Mi, Xinxue Wang, Wentao Zheng, Jian Wang, Tong Lin, Mengxia Sun, Mingli Su, Hong Li & Hua Ye

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  1. Fangxia Mi
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Mi, F., Wang, X., Zheng, W. et al. Effects of Different Preparation Methods on Microbiota Composition of Fecal Suspension. Mol Biotechnol 65, 871–880 (2023). https://doi.org/10.1007/s12033-022-00590-1

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  • DOI: https://doi.org/10.1007/s12033-022-00590-1

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