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Updated: Jan 12, 2026
An In Vitro Batch-culture Model to Estimate the Effects of Interventional Regimens on Human Fecal Microbiota
Published on: July 31, 2019
Impact of storage conditions on live bacteria in partially processed faecal microbiota transplantation products using
Katrine Agerbo Hovmand1,2,3, Flemming Damgaard Nielsen3, Emil Ainsworth Jochumsen4
1OUH Frontline Centre for Applied Microbiota and Mucosal Barrier Research (MICARE), Department of Rheumatology, Odense University Hospital, 5000 Odense, Denmark.
Abstract:
Faecal microbiota transplantation (FMT) is an effective treatment for Clostridioides difficile infection, but current FMT product manufacturing protocols are time-sensitive, labour-intensive, and require specialized staff. Delays can compromise microbial viability and product quality. Efficient storage methods for partially processed FMT material could therefore improve flexibility and streamline production. Therefore, this feasibility study evaluated the impact of storage conditions on live bacteria in partially processed donations containing glycerol as a cryoprotectant. Using culturomics, we assessed the concentration and composition of live bacteria immediately after half-way processing (reference, 0 h) or after storage at 5°C (refrigerator) or -80°C (freezer) for up to 72 h. Each FMT product was initially processed from one faecal donation provided by one of four screened donors. Storage at -80°C preserved the amount and composition of live bacteria in partially processed products. In contrast, storage at 5°C led to reduced bacterial concentrations and compositional shifts, particularly affecting the abundance of Bacteroides spp. These results highlight the potential adverse effects of refrigerator storage on bacterial viability, suggesting it may not be suitable for maintaining the quality of partially processed encapsulated FMT products. Freezer storage, however, emerged as a reliable method to preserve the content of live bacteria for at least 72 h.
Insights
Freezer storage (-80°C) effectively preserves live bacteria in partially processed faecal microbiota transplantation (FMT) products. Refrigerator storage (5°C) reduces bacterial concentration and alters composition, impacting FMT quality.
Area of Science:
- Microbiology
- Gastroenterology
- Biotechnology
Background:
- Faecal microbiota transplantation (FMT) is a key treatment for recurrent Clostridioides difficile infection.
- Current FMT manufacturing is labor-intensive, time-sensitive, and requires specialized personnel, impacting product quality and flexibility.
- Efficient storage of intermediate FMT products is needed to streamline production and maintain microbial viability.
Purpose of the Study:
- To evaluate the impact of different storage conditions on live bacteria in partially processed FMT material.
- To assess the feasibility of using glycerol-cryopreserved, partially processed FMT donations for improved manufacturing flexibility.
Main Methods:
- Feasibility study using culturomics to analyze bacterial concentration and composition.
- Partially processed FMT donations with glycerol cryoprotectant were stored at 5°C (refrigerator) and -80°C (freezer) for up to 72 hours.
- Bacterial assessment was performed immediately after processing (0 h) and after storage intervals.
Main Results:
- Storage at -80°C maintained the concentration and composition of live bacteria in partially processed FMT products.
- Storage at 5°C resulted in decreased bacterial concentrations and significant compositional shifts, notably impacting Bacteroides spp. abundance.
- Refrigerator storage negatively affected bacterial viability and product quality.
Conclusions:
- -80°C freezer storage is a reliable method for preserving live bacteria in partially processed FMT products for at least 72 hours.
- 5°C refrigerator storage is unsuitable for maintaining the quality and bacterial viability of partially processed encapsulated FMT products.
- Optimized storage protocols are crucial for enhancing the flexibility and efficiency of FMT manufacturing.
