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Long-term stability of microbiome diversity and composition in fecal samples stored in eNAT medium.
MicrobiologyOpen ( IF 3.9 ) Pub Date : 2020-05-10 , DOI: 10.1002/mbo3.1046 Rebecca R Young 1 , Kirsten Jenkins 1 , Felix Araujo-Perez 2 , Patrick C Seed 2 , Matthew S Kelly 1
MicrobiologyOpen ( IF 3.9 ) Pub Date : 2020-05-10 , DOI: 10.1002/mbo3.1046 Rebecca R Young 1 , Kirsten Jenkins 1 , Felix Araujo-Perez 2 , Patrick C Seed 2 , Matthew S Kelly 1
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Fecal samples collected for microbiome analyses are typically frozen to avoid postcollection changes in microbial composition. eNAT is a guanidine thiocyanate‐based medium that stabilizes microbial DNA and allows safe specimen handling and shipping by inactivating microorganisms. We collected fecal samples (n = 50) from children undergoing hematopoietic stem cell transplantation. We divided samples into three aliquots: (a) stored in RNAlater and immediately transferred to −80°C; (b) stored in eNAT medium and immediately transferred to −80°C; and (c) stored in eNAT medium at ambient temperature (~20°C) for 30 days prior to transfer to −80°C. Mean (standard deviation) Shannon diversity and Chao1 indices in sample aliquots were 2.05 (0.62) and 23.8 (16.6), respectively. Comparing samples frozen immediately in RNAlater to samples frozen immediately in eNAT, there were no differences in Shannon diversity (p = .51), Chao1 richness (p = .66), and overall microbiome composition (p = .99). Comparing eNAT samples frozen immediately to samples stored at ambient temperature, we identified no differences in Shannon diversity (p = .65), Chao1 richness (p = .87), and overall microbiome composition (p = .99). Storage of fecal samples in eNAT at ambient temperature for 30 days did not alter microbiome richness, diversity, or composition. eNAT may be a useful medium for fecal microbiome studies, particularly when cold chain storage is unavailable.
中文翻译：
储存在eNAT培养基中的粪便样品中微生物组多样性和组成的长期稳定性。
通常将收集用于微生物组分析的粪便样品以避免收集后微生物成分的变化。eNAT是一种基于硫氰酸胍的培养基，可稳定微生物DNA并通过灭活微生物实现安全的标本处理和运输。我们收集了粪便样本（n = 50）来自接受造血干细胞移植的儿童。我们将样品分成三等份：（a）储存在RNAlater中，并立即转移至-80°C；（b）储存在eNAT介质中，并立即转移至-80°C；（c）在转移至-80°C之前，应在环境温度（〜20°C）下保存在eNAT介质中30天。样品等分试样中的Shannon多样性平均值和Chao1指数分别为2.05（0.62）和23.8（16.6）。将立即在RNAlater中冷冻的样品与在eNAT中立即冷冻的样品进行比较，香农多样性（p = .51），Chao1丰富度（p = .66）和总体微生物组组成（p = .99）。将立即冷冻的eNAT样品与室温下储存的样品进行比较，我们发现香农多样性（p = .65），Chao1丰富度（p = .87）和整体微生物组组成（p = .99）没有差异。将粪便样品在环境温度下在eNAT中保存30天不会改变微生物组的丰富度，多样性或组成。eNAT可能是粪便微生物组研究的有用介质，尤其是在没有冷链存储时。
更新日期：2020-05-10
中文翻译：
储存在eNAT培养基中的粪便样品中微生物组多样性和组成的长期稳定性。
通常将收集用于微生物组分析的粪便样品以避免收集后微生物成分的变化。eNAT是一种基于硫氰酸胍的培养基，可稳定微生物DNA并通过灭活微生物实现安全的标本处理和运输。我们收集了粪便样本（n = 50）来自接受造血干细胞移植的儿童。我们将样品分成三等份：（a）储存在RNAlater中，并立即转移至-80°C；（b）储存在eNAT介质中，并立即转移至-80°C；（c）在转移至-80°C之前，应在环境温度（〜20°C）下保存在eNAT介质中30天。样品等分试样中的Shannon多样性平均值和Chao1指数分别为2.05（0.62）和23.8（16.6）。将立即在RNAlater中冷冻的样品与在eNAT中立即冷冻的样品进行比较，香农多样性（p = .51），Chao1丰富度（p = .66）和总体微生物组组成（p = .99）。将立即冷冻的eNAT样品与室温下储存的样品进行比较，我们发现香农多样性（p = .65），Chao1丰富度（p = .87）和整体微生物组组成（p = .99）没有差异。将粪便样品在环境温度下在eNAT中保存30天不会改变微生物组的丰富度，多样性或组成。eNAT可能是粪便微生物组研究的有用介质，尤其是在没有冷链存储时。
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