3016414154dde62c71f5e3a7c8b31c8c pmc_34945571.pdf 70ca18bd6ec99d638e685ebc519b5fc81b39e152 pmc_34945571.pdf cb66ef475bdbd388fa48b2bc14fbeb9685b510b05985544a9a3f46dbeada63e8 pmc_34945571.pdf Title: An In Vitro Protocol to Study the Modulatory Effects of a Food or Biocompound on Human Gut Microbiome and Metabolome Subject: The gut microbiota plays a key role in gastrointestinal immune and metabolic functions and is influenced by dietary composition. An in vitro protocol simulating the physiological conditions of the digestive system helps to study the effects of foods/biocompounds on gut microbiome and metabolome. The Dynamic-Colonic Gastrointestinal Digester consists of five interconnected compartments, double jacket vessels that simulate the physiological conditions of the stomach, the small intestine and the three colonic sections, which are the ascending colon, transverse colon and descending colon. Human faeces are required to reproduce the conditions and culture medium of the human colon, allowing the growth of the intestinal microbiota. After a stabilization period of 12 days, a food/biocompound can be introduced to study its modulatory effects during the next 14 days (treatment period). At the end of the stabilization and treatment period, samples taken from the colon compartments are analysed. The 16S rRNA gene analysis reveals the microbiota composition. The untargeted metabolomics analysis gives more than 10,000 features (metabolites/compounds). The present protocol allows in vitro testing of the modulatory effects of foods or biocompounds on gut microbiota composition and metabolic activity. Keywords: microbiota; digestor; food compounds; colonic fermentation; metabolome Author: Carles Rosés, Juan Antonio Nieto, Blanca Viadel, Elisa Gallego, Ana Romo-Hualde, Sergio Streitenberger, Fermín I. Milagro and Anna Barceló Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Mon Dec 6 11:02:32 2021 CET ModDate: Mon Dec 6 15:11:22 2021 CET Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 14 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2127626 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- ZIENDO+VnURWPalladioL Type 1 Custom yes yes no 10 0 ZBEYTI+URWPalladioL-Ital Type 1 Custom yes yes no 15 0 DPTCTD+URWPalladioL-Bold Type 1 Custom yes yes no 20 0 RELVEM+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 XWOCXD+VnURWPalladioL-Bold Type 1 Custom yes yes no 28 0 XBNVAE+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 36 0 MCBCCI+SourceSansRoman-Regular Type 1C Custom yes yes no 58 0 MCBCCM+SourceSansRoman.613wght Type 1C Custom yes yes no 62 0 MCBCCO+SourceSansRoman-Bold Type 1C Custom yes yes no 66 0 YFFKLH+CMSY10 Type 1 Builtin yes yes no 78 0 OVNEXW+EURM10 Type 1 Builtin yes yes no 85 0 ACLDKI+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 108 0 ACLDKK+PalatinoLinotype TrueType WinAnsi yes yes no 111 0 ACLDKM+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 114 0 ACLDKI+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 126 0 ACLDKK+PalatinoLinotype TrueType WinAnsi yes yes no 129 0 ACLDKM+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 132 0 ACLDKI+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 149 0 ACLDKK+PalatinoLinotype TrueType WinAnsi yes yes no 152 0 ACLDKM+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 155 0 ACLDKI+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 172 0 ACLDKK+PalatinoLinotype TrueType WinAnsi yes yes no 175 0 ACLDKM+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 178 0 ACLDKI+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 189 0 ACLDKK+PalatinoLinotype TrueType WinAnsi yes yes no 192 0 ACLDKM+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 195 0 ACLDKI+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 213 0 ACLDKK+PalatinoLinotype TrueType WinAnsi yes yes no 216 0 ACLDKM+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 219 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2023-05-27 02:52:57 CEST RepresentationInformation: pmc_34945571.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2023-03-31 12:06:36 CEST Size: 2127626 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 372 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@44c8afef Item: Title: Material and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Children: Item: Title: In Vitro Digestion Models Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Item: Title: Description of the Dynamic Gastrointestinal and Colonic Fermentation Model Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Faecal Inoculum Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: Process Description and Duration Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: Description and Volume of the Product to Be Tested Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: Short Fatty Acid Analyses Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: DNA Extraction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: Metagenomic Data: Library Preparation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: Data Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: Metagenomic Data: Analysis and Processing Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Richness and Evenness Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Item: Title: Untargeted Metabolomics Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: Results and Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Item: Title: Study Limitations Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Info: Title: An In Vitro Protocol to Study the Modulatory Effects of a Food or Biocompound on Human Gut Microbiome and Metabolome Author: Carles Rosés, Juan Antonio Nieto, Blanca Viadel, Elisa Gallego, Ana Romo-Hualde, Sergio Streitenberger, Fermín I. Milagro and Anna Barceló Subject: The gut microbiota plays a key role in gastrointestinal immune and metabolic functions and is influenced by dietary composition. An in vitro protocol simulating the physiological conditions of the digestive system helps to study the effects of foods/biocompounds on gut microbiome and metabolome. The Dynamic-Colonic Gastrointestinal Digester consists of five interconnected compartments, double jacket vessels that simulate the physiological conditions of the stomach, the small intestine and the three colonic sections, which are the ascending colon, transverse colon and descending colon. Human faeces are required to reproduce the conditions and culture medium of the human colon, allowing the growth of the intestinal microbiota. After a stabilization period of 12 days, a food/biocompound can be introduced to study its modulatory effects during the next 14 days (treatment period). At the end of the stabilization and treatment period, samples taken from the colon compartments are analysed. The 16S rRNA gene analysis reveals the microbiota composition. The untargeted metabolomics analysis gives more than 10,000 features (metabolites/compounds). The present protocol allows in vitro testing of the modulatory effects of foods or biocompounds on gut microbiota composition and metabolic activity. 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