title: Microbial Metabolism Shifts Towards an Adverse Profile with Supplementary Iron in the TIM-2 In vitro Model of the Human Colon.
abstract: Oral iron administration in African children can increase the risk for infections. However, it remains unclear to what extent supplementary iron affects the intestinal microbiome. We here explored the impact of iron preparations on microbial growth and metabolism in the well-controlled TNO's in vitro model of the large intestine (TIM-2). The model was inoculated with a human microbiota, without supplementary iron, or with 50 or 250 μmol/L ferrous sulfate, 50 or 250 μmol/L ferric citrate, or 50 μmol/L hemin. High resolution responses of the microbiota were examined by 16S rDNA pyrosequencing, microarray analysis, and metagenomic sequencing. The metabolome was assessed by fatty acid quantification, gas chromatography-mass spectrometry (GC-MS), and (1)H-NMR spectroscopy. Cultured intestinal epithelial Caco-2 cells were used to assess fecal water toxicity. Microbiome analysis showed, among others, that supplementary iron induced decreased levels of Bifidobacteriaceae and Lactobacillaceae, while it caused higher levels of Roseburia and Prevotella. Metagenomic analyses showed an enrichment of microbial motility-chemotaxis systems, while the metabolome markedly changed from a saccharolytic to a proteolytic profile in response to iron. Branched chain fatty acids and ammonia levels increased significantly, in particular with ferrous sulfate. Importantly, the metabolite-containing effluent from iron-rich conditions showed increased cytotoxicity to Caco-2 cells. Our explorations indicate that in the absence of host influences, iron induces a more hostile environment characterized by a reduction of microbes that are generally beneficial, and increased levels of bacterial metabolites that can impair the barrier function of a cultured intestinal epithelial monolayer.
authors.affiliation: Department of Laboratory Medicine - Translational Metabolic Laboratory, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center Nijmegen, Netherlands.
authors.forename: Guus A M
authors.initials: GA
authors.lastname: Kortman
authors.affiliation: Radboud Institute for Molecular Life Sciences, Center for Molecular and Biomolecular Informatics, Radboud University Medical CenterNijmegen, Netherlands; Theoretical Biology and Bioinformatics, Utrecht UniversityUtrecht, Netherlands; Department of Marine Biology, Institute of Biology, Federal University of Rio de JaneiroRio de Janeiro, Brazil.
authors.forename: Bas E
authors.initials: BE
authors.lastname: Dutilh
authors.affiliation: TNO Innovation for Life Zeist, Netherlands.
authors.forename: Annet J H
authors.initials: AJ
authors.lastname: Maathuis
authors.affiliation: Department of Laboratory Medicine - Translational Metabolic Laboratory, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center Nijmegen, Netherlands.
authors.forename: Udo F
authors.initials: UF
authors.lastname: Engelke
authors.affiliation: NIZO food research, Kernhemseweg Ede Netherlands.
authors.forename: Jos
authors.initials: J
authors.lastname: Boekhorst
authors.affiliation: Argonne National Laboratory, University of Chicago Lemont, IL, USA.
authors.forename: Kevin P
authors.initials: KP
authors.lastname: Keegan
authors.affiliation: Illumina Inc., Chesterford Research ParkLittle Chesterford, UK; DNAdigest, Future Business CentreCambridge, UK.
authors.forename: Fiona G G
authors.initials: FG
authors.lastname: Nielsen
authors.affiliation: Illumina Inc., Chesterford Research Park Little Chesterford, UK.
authors.forename: Jason
authors.initials: J
authors.lastname: Betley
authors.affiliation: Illumina Inc., Chesterford Research Park Little Chesterford, UK.
authors.forename: Jacqueline C
authors.initials: JC
authors.lastname: Weir
authors.affiliation: Illumina Inc., Chesterford Research Park Little Chesterford, UK.
authors.forename: Zoya
authors.initials: Z
authors.lastname: Kingsbury
authors.affiliation: Department of Laboratory Medicine - Translational Metabolic Laboratory, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center Nijmegen, Netherlands.
authors.forename: Leo A J
authors.initials: LA
authors.lastname: Kluijtmans
authors.affiliation: Department of Laboratory Medicine - Translational Metabolic Laboratory, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center Nijmegen, Netherlands.
authors.forename: Dorine W
authors.initials: DW
authors.lastname: Swinkels
authors.affiliation: TNO Innovation for Life Zeist, Netherlands.
authors.forename: Koen
authors.initials: K
authors.lastname: Venema
authors.affiliation: Department of Laboratory Medicine - Translational Metabolic Laboratory, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center Nijmegen, Netherlands.
authors.forename: Harold
authors.initials: H
authors.lastname: Tjalsma
keywords: BCFA; SCFA; gut microbiome; in vitro fermentation; iron supplementation; metabolome; metagenome; toxicity