Shaheen Mohamed, McDougall Chelsea, Chan Leona, Franz Rose, Wong Karen, Giebelhaus Ryland T, Nguyen Gwen, Nam Seo Lin, de la Mata A Paulina, Yeo Sam, Harynuk James J, Pakpour Sepideh, Xu Huiping, Kao Dina
Division of Gastroenterology, Department of Medicine, University of Alberta, Edmonton, AB T6G 2X8, Canada.
Department of Chemistry, University of Alberta, Edmonton, AB T6G 2G2, Canada.
Microorganisms. 2025 Mar 4;13(3):587. doi: 10.3390/microorganisms13030587.
Fecal microbiota transplantation (FMT) is the most effective therapy for preventing recurrent infection (rCDI). However, the impact of FMT formulations and storage conditions on bacterial viability, community structure, functionality, and clinical efficacy remains under-investigated. We studied the effect of different storage conditions on the bacterial viability (live/dead staining and cell sorting), community structure (16S rDNA analysis), and metabolic functionality (fermentation) of frozen and lyophilized FMT formulations. The clinical success rates of rCDI patients were correlated retrospectively with FMT formulations, storage durations, and host factors using the Edmonton FMT program database. Bacterial viability remained at 10-20% across various storage conditions and formulations and was comparable to that of fresh FMT. Live and dead bacterial fractions in both frozen and lyophilized FMT preparations exhibited distinct community structures. Storage durations, but not temperatures, negatively affected bacterial diversity. More short-chain fatty acids were found in the metabolomic profiling of in vitro fermentation products using lyophilized than frozen FMT. Clinical success rates in 537 rCDI patients receiving a single dose of FMT were not significantly different among the three formulations. However, longer storage durations and advanced recipient age negatively impacted clinical efficacy. Together, our findings suggest that FMT formulations and storage durations should be considered when establishing guidelines for product shelf life for optimal treatment outcomes.
粪便微生物群移植（FMT）是预防复发性艰难梭菌感染（rCDI）最有效的疗法。然而，FMT制剂和储存条件对细菌活力、群落结构、功能及临床疗效的影响仍研究不足。我们研究了不同储存条件对冷冻和冻干FMT制剂的细菌活力（活/死染色及细胞分选）、群落结构（16S rDNA分析）和代谢功能（发酵）的影响。利用埃德蒙顿FMT项目数据库，对rCDI患者的临床成功率与FMT制剂、储存时间和宿主因素进行回顾性关联分析。在各种储存条件和制剂中，细菌活力均保持在10%-20%，与新鲜FMT相当。冷冻和冻干FMT制剂中的活菌和死菌部分均呈现出不同的群落结构。储存时间而非温度对细菌多样性有负面影响。在使用冻干FMT而非冷冻FMT的体外发酵产物代谢组分析中发现了更多短链脂肪酸。在接受单剂量FMT的537例rCDI患者中，三种制剂的临床成功率无显著差异。然而，较长的储存时间和较高的受者年龄对临床疗效有负面影响。总之，我们的研究结果表明，在制定产品保质期指南以获得最佳治疗效果时，应考虑FMT制剂和储存时间。
