Journal
NUTRIENTS
Volume 14, Issue 16, Pages -Publisher
MDPI
DOI: 10.3390/nu14163315
Keywords
gut microbiome; diaper; toddlers; epidemiology; feces
Categories
Funding
- Japan Society for the Promotion of Science (JSPS) - Ministry of the Environment, Japan [19H01078, 19K24288, 20K13824]
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The composition of human gut microbiota has a long-term impact on human health and disease. This study validated two methods for stool specimen collection and storage, and found them to be useful in large-scale epidemiological studies. However, these methods may alter the relative abundances of certain bacteria.
The composition of human gut microbiota influences human health and disease over the long term. Since the flora in specimens can easily change at ambient temperature outside the body, epidemiological studies need feasible methods of stool specimen collection and storage to be established. We aimed to validate two methods: feces frozen-stored in tubes containing guanidine thiocyanate solution for two months after collection (Method B), and feces excreted in diapers and frozen-stored (Method C). Validation was by comparison with a gold standard Method A. Bacterial flora of five adults were sampled and stored by all three methods. Bacterial composition was examined by amplicon sequencing analysis. Bland-Altman analyses showed that Methods B and C might change relative abundances of certain bacterial flora. Thereafter, we analyzed the bacterial flora of 76 toddlers (two age groups) in stools sampled and processed by Method C. The diversity indices of toddlers' flora were less than those of adults. The relative abundance of some bacteria differed significantly between children aged 1.5 and 3 years. The specimen collection and storage methods validated in this study are worth adopting in large-scale epidemiological studies, especially for small children, provided the limited accuracy for some specific bacteria is understood.
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