Protocol for fecal microbiota transplantation: A microaerophilic approach for mice housed in a specific pathogen-free facility

Shari Wouters; Hugo Moors; Mieke Verslegers; Natalie Leys; Surbhi Malhotra-Kumar; Samir Kumar-Singh; Mohamed Mysara

doi:10.1016/j.xpro.2024.103517

Protocol for fecal microbiota transplantation: A microaerophilic approach for mice housed in a specific pathogen-free facility

STAR Protoc. 2025 Jan 7;6(1):103517. doi: 10.1016/j.xpro.2024.103517. Online ahead of print.

Authors

Shari Wouters¹, Hugo Moors², Mieke Verslegers², Natalie Leys², Surbhi Malhotra-Kumar³, Samir Kumar-Singh⁴, Mohamed Mysara⁵

Affiliations

¹ Microbiology & Radiobiology Units, Belgian Nuclear Research Centre SCK CEN, 2400 Mol, Belgium; Molecular Pathology Group, Cell Biology and Histology, Faculty of Medicine and Health Sciences, University of Antwerp, 2000 Antwerp, Belgium. Electronic address: [email protected].
² Microbiology & Radiobiology Units, Belgian Nuclear Research Centre SCK CEN, 2400 Mol, Belgium.
³ Laboratory of Medical Microbiology, Vaccine and Infection Disease Institute, Faculty of Medicine and Health Sciences, University of Antwerp, 2000 Antwerp, Belgium.
⁴ Molecular Pathology Group, Cell Biology and Histology, Faculty of Medicine and Health Sciences, University of Antwerp, 2000 Antwerp, Belgium.
⁵ Microbiology & Radiobiology Units, Belgian Nuclear Research Centre SCK CEN, 2400 Mol, Belgium; Bioinformatics Group, Center for Informatics Science, Nile University, Giza, Egypt. Electronic address: [email protected].

PMID: 39772388
DOI: 10.1016/j.xpro.2024.103517

Abstract

Recently, studies have emerged exploring the potential application of fecal microbiota transplantation (FMT) in pre-clinical settings. Here, we present a protocol for FMT for mice housed in a specific pathogen-free (SPF) facility. We describe steps for sample collection, microaerophilic processing of freshly collected fecal pellets, and administration through oral gavage. We then detail procedures for the engraftment of the bacterial community. This protocol focuses on age- and gender-matched, healthy donor mice using a mobile and cost-effective alternative to an anoxic cabinet.

Keywords: health sciences; microbiology; model organisms; sequence analysis; systems biology.