{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,10]],"date-time":"2026-04-10T17:17:56Z","timestamp":1775841476509,"version":"3.50.1"},"reference-count":44,"publisher":"MDPI AG","issue":"16","license":[{"start":{"date-parts":[[2022,8,12]],"date-time":"2022-08-12T00:00:00Z","timestamp":1660262400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Grants-in-Aid for Scientific Research","award":["19H01078"],"award-info":[{"award-number":["19H01078"]}]},{"name":"Grants-in-Aid for Scientific Research","award":["19K24288"],"award-info":[{"award-number":["19K24288"]}]},{"name":"Grants-in-Aid for Scientific Research","award":["20K13824"],"award-info":[{"award-number":["20K13824"]}]},{"name":"Ministry of the Environment, Japan","award":["19H01078"],"award-info":[{"award-number":["19H01078"]}]},{"name":"Ministry of the Environment, Japan","award":["19K24288"],"award-info":[{"award-number":["19K24288"]}]},{"name":"Ministry of the Environment, Japan","award":["20K13824"],"award-info":[{"award-number":["20K13824"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nutrients"],"abstract":"<jats:p>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\u2013Altman 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\u2019 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.<\/jats:p>","DOI":"10.3390\/nu14163315","type":"journal-article","created":{"date-parts":[[2022,8,15]],"date-time":"2022-08-15T03:33:20Z","timestamp":1660534400000},"page":"3315","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Epidemiological Studies of Children\u2019s Gut Microbiota: Validation of Sample Collection and Storage Methods and Microbiota Analysis of Toddlers\u2019 Feces Collected from Diapers"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7410-0184","authenticated-orcid":false,"given":"Hazuki","family":"Tamada","sequence":"first","affiliation":[{"name":"Department of Occupational and Environmental Health, Graduate School of Medical Sciences, Nagoya City University, Mizuho-ku, Nagoya 4678601, Aichi, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1617-1595","authenticated-orcid":false,"given":"Yuki","family":"Ito","sequence":"additional","affiliation":[{"name":"Department of Occupational and Environmental Health, Graduate School of Medical Sciences, Nagoya City University, Mizuho-ku, Nagoya 4678601, Aichi, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3342-1058","authenticated-orcid":false,"given":"Takeshi","family":"Ebara","sequence":"additional","affiliation":[{"name":"Department of Occupational and Environmental Health, Graduate School of Medical Sciences, Nagoya City University, Mizuho-ku, Nagoya 4678601, Aichi, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2527-3964","authenticated-orcid":false,"given":"Sayaka","family":"Kato","sequence":"additional","affiliation":[{"name":"Department of Occupational and Environmental Health, Graduate School of Medical Sciences, Nagoya City University, Mizuho-ku, Nagoya 4678601, Aichi, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1580-9748","authenticated-orcid":false,"given":"Kayo","family":"Kaneko","sequence":"additional","affiliation":[{"name":"Department of Occupational and Environmental Health, Graduate School of Medical Sciences, Nagoya City University, Mizuho-ku, Nagoya 4678601, Aichi, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2935-3064","authenticated-orcid":false,"given":"Taro","family":"Matsuki","sequence":"additional","affiliation":[{"name":"Department of Occupational and Environmental Health, Graduate School of Medical Sciences, Nagoya City University, Mizuho-ku, Nagoya 4678601, Aichi, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2265-377X","authenticated-orcid":false,"given":"Mayumi","family":"Sugiura-Ogasawara","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, Graduate School of Medical Sciences, Nagoya City University, Mizuho-ku, Nagoya 4678601, Aichi, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6911-3351","authenticated-orcid":false,"given":"Shinji","family":"Saitoh","sequence":"additional","affiliation":[{"name":"Department of Pediatrics and Neonatology, Graduate School of Medical Sciences, Nagoya City University, Mizuho-ku, Nagoya 4678601, Aichi, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0670-8790","authenticated-orcid":false,"given":"Michihiro","family":"Kamijima","sequence":"additional","affiliation":[{"name":"Department of Occupational and Environmental Health, Graduate School of Medical Sciences, Nagoya City University, Mizuho-ku, Nagoya 4678601, Aichi, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,8,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1262","DOI":"10.1126\/science.1223813","article-title":"Host-gut microbiota metabolic interactions","volume":"336","author":"Nicholson","year":"2012","journal-title":"Science"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"104","DOI":"10.3389\/fcimb.2012.00104","article-title":"The function of our microbiota: Who is out there and what do they do?","volume":"2","author":"Ottman","year":"2012","journal-title":"Front. 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