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粪便冷冻保存期影响粪便微生物群移植的定植能力。

Faecal freezing preservation period influences colonization ability for faecal microbiota transplantation.

机构信息

Department of Gastroenterology, Juntendo University School of Medicine, Bunkyo-ku, Tokyo, Japan.

Animal Research Center, Sapporo Medical University School of Medicine, Chuo-ku, Sapporo, Japan.

出版信息

J Appl Microbiol. 2019 Mar;126(3):973-984. doi: 10.1111/jam.14167. Epub 2019 Jan 7.

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.

摘要

目的

粪便微生物群移植(FMT)作为一种治疗方法受到越来越多的关注。虽然冷冻保存于-20°C 的供体粪便由于实际优势而被广泛应用,但-20°C 的冷冻会影响细菌的活力。为了改进 FMT 的方法学,有必要对新鲜和冷冻粪便作为移植物的充分性进行评估。

方法和结果

通过使用吖啶橙单染色(PMA 微生物组)对新鲜和冷冻条件下粪便标本中的活菌组成进行了微生物组分析,比较了新鲜和冷冻条件下粪便标本中的活菌组成。此外,使用小鼠模型比较了新鲜和冷冻 FMT 中抗生素减少细菌后的恢复能力。PMA 微生物组的结果表明,冷冻和冻融处理不会显著影响体外粪便细菌的活力。然而,冷冻 FMT 中从抗菌清洗中恢复的效果会随着冷冻时间的延长而降低,尤其是在放线菌门和拟杆菌门。

结论

与 1 个月-20°C 保存相比,粪便的短期冷冻保存显示冷冻 FMT 中保留了肠道定植能力。

研究的意义和影响

移植粪便在-20°C 下长期保存可能导致 FMT 治疗的临床结果不稳定。根据疾病类型对 FMT 治疗的实际操作程序进行标准化是可取的。

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