P862 Colonisation of viable bacteroidetes in fresh or frozen faecal microbiota transplantation

• Published in: Journal of Crohn's and colitis 2018-01, Vol.12 (supplement_1), p.S552-S553
• Main Authors: Takahashi, M, Ishikawa, D, Kamei, M, Shibuya, T, Osada, T, Nagahara, A
• Format: Article
• Language: English
• ISSN: 1873-9946; 1876-4479
• DOI: 10.1093/ecco-jcc/jjx180.989

Abstract Background Fecal microbiota transplantation (FMT) with using both fresh and frozen fecal samples is an effective treatment for recurrent Clostridium difficile infection, whereas there are no established standard practices for FMT for ulcerative colitis (UC); therefore, it is not known whether fresh or frozen FMT is better as transplanted stool. We previously reported that fresh FMT following the multiple antibiotic therapy (AFM: amoxicillin, fosfomycin and metronidazole) synergistically contributes to the recovery of the phylum Bacteroidetes composition, which is associated with high clinical response. Our results suggested that key bacteria involved in UC should be delivered live to the intestines, and fresh FMT was more suitable as a treatment methodology for transplantation of viable Bacteroidetes cells. Methods Frozen samples were added glycerol to the final concentration of 10% and immediately placed in a −20℃ freezer and were preserved for a day, a week and four weeks. Four times-freeze–thawed samples were taken out of the −20℃ freezer and thawed at Room Temperature for 30 min. In order to evaluate colonisation efficiency of viable bacteria, FMT was performed to murine recipients by a single dose of fecal solution under various preservation conditions via stomach gavage after the antimicrobial bowel cleansing (ampicillin, neomycin, metronidazole and vancomycin). From all recipient mice, fecal samples were collected before antibiotic treatment, before FMT and two weeks post FMT. To analyse just only living bacteria, we validated a 16S rRNA metagenome method targeting viable cell DNA, which was based on a viability PCR technique through the use of propidium monoazide (PMA). Results The number of operational taxonomic units (OTUs) and compositions of the viable fecal bacteria at order level did not change significantly even by four times-freeze–thawed method and cryopreservation for four weeks. Whereas, in FMT for murine models, there was significant difference in the number of OTUs of the viable fecal bacteria between fresh FMT and frozen FMT for four weeks (n = 3, 3; p < 0.05). At two weeks post FMT, the total average of relative abundances of Bacteroidetes significantly decreased in those for frozen faces for four weeks compared with in using fresh faces (Relative abundance; n = 6, 6; frozen: 2.0 ± 0.7% vs. fresh: 4.9 ± 2.1%; p = 0.01). Conclusions No remarkable microbial alteration by a freezing treatment was found in vitro viability of fac