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Reproducible chemostat cultures to minimize eukaryotic viruses from fecal transplant material
Signe Adamberg1, Torben Sølbeck Rasmussen2, Sabina Brigitte Larsen2
1Department of Chemistry and Biotechnology, Tallinn University of Technology, Akadeemia tee 15, Tallinn, Estonia.
Abstract:
Recent studies indicate an important role of bacteriophages for successful fecal microbiota transplantation (FMT). However, wider clinical applications of FMT are hampered by to donor variability and concerns of infection risks by bacteria and human viruses. To overcome these challenges, mouse cecal and human fecal material were propagated in a chemostat fermentation setup supporting multiplication of bacteria, and phages, while propagation of eukaryotic viruses will be prevented in the absence of eukaryotic host cells. The results showed decrease of the median relative abundance of viral contigs of classified eukaryotic viruses below 0.01%. The corresponding virome profiles showed dilution rate dependency, a reproducibility between biological replicates, and maintained high diversity regarding both the human and mouse inocula. This proof-of-concept cultivation approach may constitute the first step of developing novel therapeutic tools with high reproducibility and with low risk of infection from the donor material to target gut-related diseases.
Insights
Fecal microbiota transplantation (FMT) can be improved using a novel cultivation method. This approach enhances reproducibility and reduces infection risks from viruses, paving the way for safer gut disease therapies.
Area of Science:
- Microbiology
- Virology
- Gastroenterology
Background:
- Bacteriophages are crucial for fecal microbiota transplantation (FMT) success.
- Clinical FMT applications are limited by donor variability and infection risks (bacteria, viruses).
Purpose of the Study:
- To develop a cultivation method for FMT preparation that reduces eukaryotic virus contamination.
- To assess the impact of this method on bacterial and phage populations and overall virome diversity.
Main Methods:
- Propagating mouse cecal and human fecal material in a chemostat fermentation system.
- The system was designed to support bacteria and phage growth while preventing eukaryotic virus propagation.
- Virome profiles were analyzed to assess viral abundance and diversity.
Main Results:
- A significant decrease in the relative abundance of eukaryotic viruses (below 0.01%) was observed.
- Virome profiles demonstrated dilution rate dependency and reproducibility across biological replicates.
- High diversity of human and mouse inocula was maintained.
Conclusions:
- This chemostat cultivation approach offers a proof-of-concept for safer, more reproducible FMT preparations.
- It may lead to novel therapeutic tools for gut-related diseases with reduced infection risks.
