A Comparison of Methods of Gut Microbiota Transplantation for Preclinical Studies

Int J Mol Sci. 2023 Jul 26;24(15):12005. doi: 10.3390/ijms241512005.

Abstract

The experimental details reported in preclinical fecal microbiota transplantation (FMT) protocols are highly inconsistent, variable, and/or incomplete. We therefore evaluated FMT from a human donor to antibiotic-induced microbial-depleted mice by exploring the effects of six techniques based on antibiotic (AB) or antibiotic + antimycotic (AB + T) gut decontamination, different administration routes, and different dosing intervals on the gut microbial population, assessed using 16S and 18S sequencing. In addition, we explored the effectiveness of FMT in terms of inflammation, physiological, and behavioral outcomes. Our results showed that intrarectal FMT at low dosing intervals better preserved the donor's gut bacterial community at genus level. Furthermore, we showed a lower abundance of several genera of fungi in animals treated with AB + T. In addition, we observed that AB + T gut decontamination followed by per os FMT, once every 3 days, affected behavioral parameters when compared to other FMT techniques. Accordingly, the same FMT groups that showed an association with some of the behavioral tests were also related to specific gut fungal genera, suggesting a possible mediation. Our findings may be useful for optimizing the practice of FMT and also in terms of donor microbiota preservation. This information may help to improve the reproducibility and reliability of FMT studies.

Keywords: 16S sequencing; 18S sequencing; antibiotic; fecal microbiota transplantation; gut microbiota; gut mycobiota.

MeSH terms

  • Animals
  • Anti-Bacterial Agents
  • Fecal Microbiota Transplantation / methods
  • Feces / microbiology
  • Gastrointestinal Microbiome*
  • Humans
  • Mice
  • RNA, Ribosomal, 16S / genetics
  • Reproducibility of Results

Substances

  • RNA, Ribosomal, 16S
  • Anti-Bacterial Agents