8baf283e8240b1e83224ae9e36015531 ijmolsci_a2021v22n2p500.pdf 3b1714d3bb9170390f45e1a8c65683e734c16807 ijmolsci_a2021v22n2p500.pdf 08fa7545b9d477d11997cfb472bb4f3bd2cabe454a86e6e93c9d0dc875898d04 ijmolsci_a2021v22n2p500.pdf Title: Effects of Rifaximin on Luminal and Wall-Adhered Gut Commensal Microbiota in Mice Subject: Rifaximin is a broad-spectrum antibiotic that ameliorates symptomatology in inflammatory/functional gastrointestinal disorders. We assessed changes in gut commensal microbiota (GCM) and Toll-like receptors (TLRs) associated to rifaximin treatment in mice. Adult C57BL/6NCrl mice were treated (7/14 days) with rifaximin (50/150 mg/mouse/day, PO). Luminal and wall-adhered ceco-colonic GCM were characterized by fluorescent in situ hybridization (FISH) and microbial profiles determined by terminal restriction fragment length polymorphism (T-RFLP). Colonic expression of TLR2/3/4/5/7 and immune-related markers was assessed (RT-qPCR). Regardless the period of treatment or the dose, rifaximin did not alter total bacterial counts or bacterial biodiversity. Only a modest increase in Bacteroides spp. (150 mg/1-week treatment) was detected. In control conditions, only Clostridium spp. and Bifidobacterium spp. were found attached to the colonic epithelium. Rifaximin showed a tendency to favour their adherence after a 1-week, but not 2-week, treatment period. Minor up-regulation in TLRs expression was observed. Only the 50 mg dose for 1-week led to a significant increase (by 3-fold) in TLR-4 expression. No changes in the expression of immune-related markers were observed. Rifaximin, although its antibacterial properties, induces minor changes in luminal and wall-adhered GCM in healthy mice. Moreover, no modulation of TLRs or local immune systems was observed. These findings, in normal conditions, do not rule out a modulatory role of rifaximin in inflammatory and or dysbiotic states of the gut. Keywords: dysbiosis; gut commensal microbiota; host-bacterial interaction systems; immune markers; rifaximin; toll-like receptors Author: Marina Ferrer, Mònica Aguilera and Vicente Martinez Creator: LaTeX with hyperref package Producer: pdfTeX-1.40.18 CreationDate: Fri Jan 8 03:32:09 2021 CET ModDate: Fri Jan 8 03:37:33 2021 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 19 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2187505 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- NBMIUS+VnURWPalladioL Type 1 Custom yes yes no 10 0 EAYRGS+URWPalladioL-Ital Type 1 Custom yes yes no 15 0 EGLQGB+URWPalladioL-Bold Type 1 Custom yes yes no 20 0 HUYOUY+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 WLFTRW+VnURWPalladioL-Bold Type 1 Custom yes yes no 28 0 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KDPFLG+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 555 0 KDPFNH+PalatinoLinotype TrueType WinAnsi yes yes no 558 0 KDPFNJ+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 561 0 KDPJJF+Arial CID TrueType Identity-H yes yes yes 564 0 MPUDFL+PazoMath Type 1 Builtin yes yes no 657 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-10-02 03:56:58 CEST RepresentationInformation: ijmolsci_a2021v22n2p500.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2023-03-31 11:50:18 CEST Size: 2187505 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 911 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Results Destination: section.2 Children: Item: Title: Effects of Rifaximin on Body Weight, Weight of Body Organs and Colonic Histology Destination: subsection.2.1 Item: Title: Effects of Rifaximin on Luminal GCM Destination: subsection.2.2 Item: Title: Effects of Rifaximin on Bacterial Adherence to the Colonic Wall Destination: subsection.2.3 Item: Title: Effects of Rifaximin on Colonic Expression of TLRs and Immune-Related Markers Destination: subsection.2.4 Item: Title: Discussion Destination: section.3 Item: Title: Materials and Methods Destination: section.4 Children: Item: Title: Animals Destination: subsection.4.1 Item: Title: Antibiotic Destination: subsection.4.2 Item: Title: Experimental Protocols Destination: subsection.4.3 Item: Title: Samples Collection Destination: subsection.4.4 Item: Title: Histological Evaluation Destination: subsection.4.5 Item: Title: Bacterial Identification by Fluorescence in Situ Hybridization (FISH) Destination: subsection.4.6 Item: Title: Terminal Restriction Fragment Length Polymorphism Destination: subsection.4.7 Item: Title: Colonic Expression of TLRs and Immune-Related Markers Using Quantitative Real-Time PCR (RT-qPCR) Destination: subsection.4.8 Item: Title: Statistical Analysis Destination: subsection.4.9 Item: Title: References Destination: subsection.4.9 Info: Title: Effects of Rifaximin on Luminal and Wall-Adhered Gut Commensal Microbiota in Mice Author: Marina Ferrer, Mònica Aguilera and Vicente Martinez Subject: Rifaximin is a broad-spectrum antibiotic that ameliorates symptomatology in inflammatory/functional gastrointestinal disorders. We assessed changes in gut commensal microbiota (GCM) and Toll-like receptors (TLRs) associated to rifaximin treatment in mice. Adult C57BL/6NCrl mice were treated (7/14 days) with rifaximin (50/150 mg/mouse/day, PO). Luminal and wall-adhered ceco-colonic GCM were characterized by fluorescent in situ hybridization (FISH) and microbial profiles determined by terminal restriction fragment length polymorphism (T-RFLP). Colonic expression of TLR2/3/4/5/7 and immune-related markers was assessed (RT-qPCR). Regardless the period of treatment or the dose, rifaximin did not alter total bacterial counts or bacterial biodiversity. Only a modest increase in Bacteroides spp. (150 mg/1-week treatment) was detected. In control conditions, only Clostridium spp. and Bifidobacterium spp. were found attached to the colonic epithelium. Rifaximin showed a tendency to favour their adherence after a 1-week, but not 2-week, treatment period. Minor up-regulation in TLRs expression was observed. Only the 50 mg dose for 1-week led to a significant increase (by 3-fold) in TLR-4 expression. No changes in the expression of immune-related markers were observed. Rifaximin, although its antibacterial properties, induces minor changes in luminal and wall-adhered GCM in healthy mice. Moreover, no modulation of TLRs or local immune systems was observed. These findings, in normal conditions, do not rule out a modulatory role of rifaximin in inflammatory and or dysbiotic states of the gut. 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