Facchin Sonia, Cardin Romilda, Patuzzi Ilaria, Carlotto Chiara, Minotto Milena, Barberio Brigida, Zingone Fabiana, Besutti Valeria M, Castagliuolo Ignazio, Cattelan Annanaria, Savarino Edoardo V
Department of Surgery, Oncology and Gastroenterology (DISCOG), University of Padua, Padua, Italy; Gastroenterology Unit, Azienda Ospedale Università di Padova, Padova, Italy.
Research & Development Division, EuBiome S.r.l., 35131 Padova, Italy.
Dig Liver Dis. 2025 Mar;57(3):702-706. doi: 10.1016/j.dld.2024.11.025. Epub 2024 Dec 13.
Freezing donor fecal microbiota has improved fecal microbiota transplantation (FMT) for recurrent C. difficile infection (CDI), achieving short-term effectiveness similar to fresh-samples. Research shows frozen fecal matter remains effective for up to 12-months at -80 °C.
To assess how long-term-freezing and thawing affect the viability, microbial composition, and clinical efficacy of frozen-stools for FMT.
Stool samples from three donors were processed into 18 aliquots, thawed at intervals over two years, and analyzed for cell viability and microbial load. Microbiota composition was assessed through 16S-sequencing, with diversity evaluated using the Shannon-index and Principal-Coordinates-Analysis based on Bray-Curtis-distance (α/β-diversity). The same donors provided fecal material for a total of 23 FMT procedures, including 15 for CDI and 8 off-label.
We found that donor stools frozen for two years contained viable bacteria comparable to fresh samples, with anaerobic and aerobic species remaining viable for 24 months. Despite a reduction in colony-forming-units, FMT was successful in 71.4 % and 100 % of the cases at one year and at the end of follow-up, respectively. Most bacterial changes occurred among anaerobic species (Blautia producta and Bifidobacterium adolescentis), increasing post-thawing. Notably, specific taxa, (C. aerofaciens and Erysipelotrichaceae_Cc115), showed significant unexplained increase.
Long-term-stool-storage enhances FMT accessibility without compromising its success, despite taxonomic changes after 24 months.
冷冻供体粪便微生物群改善了复发性艰难梭菌感染（CDI）的粪便微生物群移植（FMT），实现了与新鲜样本相似的短期疗效。研究表明，冷冻粪便物质在-80°C下长达12个月仍保持有效。
评估长期冷冻和解冻对用于FMT的冷冻粪便的活力、微生物组成和临床疗效的影响。
将来自三名供体的粪便样本处理成18份等分试样，在两年内间隔解冻，并分析细胞活力和微生物负荷。通过16S测序评估微生物群组成，使用香农指数和基于布雷-柯蒂斯距离的主坐标分析（α/β多样性）评估多样性。相同的供体总共提供了23次FMT程序的粪便材料，包括15次用于CDI和8次用于非标签用途。
我们发现，冷冻两年的供体粪便含有与新鲜样本相当的活细菌，厌氧和好氧物种在24个月内仍保持活力。尽管菌落形成单位减少，但FMT在一年时和随访结束时分别有71.4%和100%的病例成功。大多数细菌变化发生在厌氧物种（产丁酸布劳特氏菌和青春双歧杆菌）之间，解冻后增加。值得注意的是，特定分类群（产气荚膜梭菌和丹毒丝菌科_Cc115）出现了显著的、无法解释的增加。
长期粪便储存提高了FMT的可及性，而不影响其成功率，尽管24个月后会有分类学变化。
