Faecal freezing preservation period influences colonization ability for faecal microbiota transplantation

Abstract

Aims There has been growing interest in faecal microbiota transplantation (FMT) as treatment. Although, frozen donor faeces preserved at −20°C has been widely used for practical advantages, freezing at −20°C can affect bacterial viability. Adequacy evaluation of fresh and frozen faeces as the transplant is necessary for the methodological improvement of FMT. Methods and Results The viable bacterial compositions of faecal specimens under fresh and freezing conditions were compared by a microbiome analysis using propidium monoazide (PMA microbiome). In addition, recovery abilities from bacterial reduction by antibiotics were compared between fresh and frozen FMT using a murine model. PMA microbiome results suggested that freezing and freeze‐thawing did not significantly affect in vitro faecal bacterial viability. However, the recovery effect from antimicrobial cleansing in frozen FMT was reduced in a freezing time‐dependent manner, especially prominent in Actinobacteria and Bacteroidetes phyla. Conclusions Short‐term freezing preservation of faeces exhibited maintenance of enteric colonization ability in frozen FMT in comparison to 1 month −20°C‐preservation. Significance and Impact of the Study Long‐term −20°C‐preservation of transplanted faeces can result in instability of the clinical outcome in FMT therapy. The standardization of practical procedures of FMT therapy according to disease types is desirable.

Changes in Microbiota Profiles After Prolonged Frozen Storage of Stool Suspensions

“…Recently, Takahashi et al (2019) showed that in a murine model, enteric colonization by Bacteroidetes, but not that of Firmicutes, was reduced by 1-month frozen storage of a fecal suspension, which is consistent with a higher sensitivity of Bacteroidetes to stressful conditions, as suggested in the present study. Interestingly, in the study of Takahashi et al, in vitro bacterial viability, assessed by propidium monoazide (PMA) 16S community profiling, was not significantly changed during 1month cryopreservation.…”

From Donor to Patient: Collection, Preparation and Cryopreservation of Fecal Samples for Fecal Microbiota Transplantation

“…Refreezing once defrosted is not permitted. Recently it has been shown that long-term preservation of transplanted feces at −20 • C can result in instability (especially concerning Actinobacteria and Bacteroidetes phyla) of the clinical outcome in FMT therapy [52].…”

Fecal Microbiota Transplantation: A Potential Tool for Treatment of Human Female Reproductive Tract Diseases

“…Laboratorists check for the presence of any pathogens, such as HIV, HBV, HCV, Treponema pallidum , Human T-lymphotropic virus I and II, Plasmodium spp., Trypanosoma spp., Mycobacterium tuberculosis, Campylobacter spp., Escherichia coli O157 H7, Yersinia , vancomycin-resistant enterococci (VRE), methicillin-resistant Staphylococcus aureus (MRSA), Gram-negative multidrug-resistant bacteria, Norovirus , antigens and/or acid fast staining for Giardia lamblia and Cryptosporidium parvum , protozoa (including Blastocystis hominis ), helminths, and fecal occult blood testing. Regarding the laboratory preparation method, it is possible to choose between two different procedures: fresh and frozen preparations (22, 23). The first procedure requires that the feces of the donor be processed within 6 h after defecation to preserve the integrity of the anaerobic bacteria (24).…”