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Home / Equine Research Bank / PeerJ / Changes in bacterial viability after preparation and storage...
PeerJ2025; 13; e18860; doi: 10.7717/peerj.18860

Changes in bacterial viability after preparation and storage of fecal microbiota transplantation solution using equine feces.

Abstract: Fecal microbiota transplantation (FMT) has been used as a treatment option for horses (Equus caballus) with gastrointestinal diseases. Several preparation and conservation protocols to improve bacterial survival have been studied in other species. Unassigned: This study aimed to evaluate the impact of oxygen exposure and different protectant solutions on bacterial viability before and after freezing using horse feces. Fecal samples from 10 healthy horses were aliquoted and diluted in cryoprotectant solutions containing antioxidants (n = 40) or 10% glycerol (n = 40). Half of the aliquots from each dilution condition were prepared inside an anaerobic chamber, while the other half were prepared under ambient air conditions. Each sample was also analyzed fresh and after freezing at -20 °C for 90 days. Bacterial viability was assessed using flow cytometry. A mixed linear model and the Friedman and Wilcoxon tests were used depending on data distribution. Unassigned: Freeze-thawing decreased bacterial viability by 47% (mean ± SD: 51 ± 27% before, 27 ± 8% after; p < 0.001). Glycerol was superior to the cryoprotectant after freezing (32 ± 8% glycerol, 24 ± 8% cryoprotectant; p < 0.001). Oxygen exposure did not affect viability (p = 0.13). There was no statistical difference between protectant solutions in fresh samples (p = 0.16). Unassigned: Fresh FMT solutions may be better for treating horses with dysbiosis, but if freezing cannot be avoided, glycerol should be used to dilute feces.
© 2025 Arantes et al.
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Publication Date: 2025-02-20 PubMed ID: 39989751PubMed Central: PMC11847485DOI: 10.7717/peerj.18860Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The study explores the changes in bacterial viability in horse feces when stored and prepared for fecal microbiota transplantation (FMT), aiming to improve treatment for horses with gastrointestinal diseases. It concluded that fresher solutions are more effective, but if freezing is necessary, the use of glycerol as a dilutant is recommended.

Objectives of the Research

  • The study was designed to investigate the impacts of oxygen exposure and diverse protectant solutions on the viability of bacteria in horse fecal matter both before and after freezing.
  • The goal was to optimize the preparation and conservation protocols for FMT, widely employed in treating horses experiencing gastrointestinal diseases.

Methodology

  • Fecal samples were obtained from 10 healthy horses and split into aliquots diluted in two types of protectant solutions, antioxidants and 10% glycerol.
  • The aliquots were exposed to two different conditions for preparation: inside an anaerobic chamber and under normal ambient air.
  • The samples were studied in two distinct states: fresh and following freezing at -20 °C for a period of 90 days.
  • Bacterial viability, the core measurement of interest, was evaluated via flow cytometry, a technique used to measure physical and chemical characteristics of cells or particles.
  • Statistical analysis was done using a mixed linear model, Friedman and Wilcoxon tests depending on data distribution.

Findings

  • The freeze-thawing process noticeably reduced bacterial viability by 47% on average.
  • Comparatively, glycerol performed better than the antioxidant-based protectant after freezing, maintaining more bacterial viability.
  • Oxygen exposure showed no significant impact on bacterial viability.
  • There was no statistically significant difference between the two protectant solutions when used on fresh samples.

Conclusions and Recommendations

  • Fresh FMT solutions are recommended for treating horses with gut dysbiosis, a condition where the normal microbial balance in the gut is disturbed.
  • However, if freezing becomes a requirement, glycerol is preferred over antioxidants as a feces dilutant.

Cite This Article

APA
Arantes JA, Di Pietro R, Ratté M, Arroyo LG, Leclère M, Costa MC. (2025). Changes in bacterial viability after preparation and storage of fecal microbiota transplantation solution using equine feces. PeerJ, 13, e18860. https://doi.org/10.7717/peerj.18860

Publication

PeerJ
ISSN: 2167-8359
NlmUniqueID: 101603425
Country: United States
Language: English
Volume: 13
Pages: e18860

Researcher Affiliations

Arantes, Julia A
  • Department of Veterinary Medicine/Faculty of Animal Science and Food Engineering, Universidade de São Paulo, Pirassununga, São Paulo, Brazil.
Di Pietro, Rebecca
  • Department of Veterinary Biomedical Sciences/Faculty of Veterinary Medicine, University of Montreal, Saint Hyacinthe, QC, Canada.
Ratté, Mélanie
  • Department of Veterinary Biomedical Sciences/Faculty of Veterinary Medicine, University of Montreal, Saint Hyacinthe, QC, Canada.
Arroyo, Luis G
  • Department of Clinical Studies/Ontario Veterinary College, University of Guelph, Guelph, ON, Canada.
Leclère, Mathilde
  • Department of Clinical Sciences/Faculty of Veterinary Medicine, University of Montreal, Saint-Hyacinthe, QC, Canada.
Costa, Marcio C
  • Department of Veterinary Biomedical Sciences/Faculty of Veterinary Medicine, University of Montreal, Saint Hyacinthe, QC, Canada.

MeSH Terms

  • Animals
  • Horses / microbiology
  • Feces / microbiology
  • Fecal Microbiota Transplantation / methods
  • Fecal Microbiota Transplantation / veterinary
  • Microbial Viability / drug effects
  • Cryoprotective Agents / pharmacology
  • Freezing
  • Glycerol / pharmacology
  • Oxygen
  • Cryopreservation

Conflict of Interest Statement

The authors declare that they have no competing interests.

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