title: An in vitro culture model to study the dynamics of colonic microbiota in Syrian golden hamsters and their susceptibility to infection with Clostridium difficile.
abstract: Clostridium difficile infections (CDI) are caused by colonization and growth of toxigenic strains of C. difficile in individuals whose intestinal microbiota has been perturbed, in most cases following antimicrobial therapy. Determination of the protective commensal gut community members could inform the development of treatments for CDI. Here, we utilized the lethal enterocolitis model in Syrian golden hamsters to analyze the microbiota disruption and recovery along a 20-day period following a single dose of clindamycin on day 0, inducing in vivo susceptibility to C. difficile infection. To determine susceptibility in vitro, spores of strain VPI 10463 were cultured with and without soluble hamster fecal filtrates and growth was quantified by quantitative PCR and toxin immunoassay. Fecal microbial population changes over time were tracked by 16S ribosomal RNA gene analysis via V4 sequencing and the PhyloChip assay. C. difficile culture growth and toxin production were inhibited by the presence of fecal extracts from untreated hamsters but not extracts collected 5 days post-administration of clindamycin. In vitro inhibition was re-established by day 15, which correlated with resistance of animals to lethal challenge. A substantial fecal microbiota shift in hamsters treated with antibiotics was observed, marked by significant changes across multiple phyla including Bacteroidetes and Proteobacteria. An incomplete return towards the baseline microbiome occurred by day 15 correlating with the inhibition of C. difficile growth in vitro and in vivo. These data suggest that soluble factors produced by the gut microbiota may be responsible for the suppression of C. difficile growth and toxin production.
authors.affiliation: Eurofins Lancaster Laboratories Inc, Lancaster, PA, USA.
authors.forename: Matthew
authors.initials: M
authors.lastname: Miezeiewski
authors.affiliation: Vaccine Research, Merck Research Laboratories, West Point, PA, USA.
authors.forename: Todd
authors.initials: T
authors.lastname: Schnaufer
authors.affiliation: Vaccine Research, Merck Research Laboratories, West Point, PA, USA.
authors.forename: Michele
authors.initials: M
authors.lastname: Muravsky
authors.affiliation: Vaccine Research, Merck Research Laboratories, West Point, PA, USA.
authors.forename: Su
authors.initials: S
authors.lastname: Wang
authors.affiliation: Vaccine Research, Merck Research Laboratories, West Point, PA, USA.
authors.forename: Ivette
authors.initials: I
authors.lastname: Caro-Aguilar
authors.affiliation: Vaccine Research, Merck Research Laboratories, West Point, PA, USA.
authors.forename: Susan
authors.initials: S
authors.lastname: Secore
authors.affiliation: Vaccine Research, Merck Research Laboratories, West Point, PA, USA.
authors.forename: David S
authors.initials: DS
authors.lastname: Thiriot
authors.affiliation: Vaccine Research, Merck Research Laboratories, West Point, PA, USA.
authors.forename: Charlie
authors.initials: C
authors.lastname: Hsu
authors.affiliation: Vaccine Research, Merck Research Laboratories, West Point, PA, USA.
authors.forename: Irene
authors.initials: I
authors.lastname: Rogers
authors.affiliation: Departments of Bioinformatics, Second Genome Inc, South San Francisco, CA, USA.
authors.forename: Todd
authors.initials: T
authors.lastname: DeSantis
authors.affiliation: Departments of Bioinformatics, Second Genome Inc, South San Francisco, CA, USA.
authors.forename: Justin
authors.initials: J
authors.lastname: Kuczynski
authors.affiliation: Departments of Bioinformatics, Second Genome Inc, South San Francisco, CA, USA.
authors.forename: Alexander J
authors.initials: AJ
authors.lastname: Probst
authors.affiliation: Departments of Bioinformatics, Second Genome Inc, South San Francisco, CA, USA.
authors.forename: Christel
authors.initials: C
authors.lastname: Chehoud
authors.affiliation: Basic Research, Second Genome Inc, South San Francisco, CA, USA.
authors.forename: Rachel
authors.initials: R
authors.lastname: Steger
authors.affiliation: Basic Research, Second Genome Inc, South San Francisco, CA, USA.
authors.forename: Janet
authors.initials: J
authors.lastname: Warrington
authors.affiliation: Vaccine Research, Merck Research Laboratories, West Point, PA, USA.
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Bodmer
authors.affiliation: Vaccine Research, Merck Research Laboratories, West Point, PA, USA.
authors.forename: Jon H
authors.initials: JH
authors.lastname: Heinrichs
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002981:Clindamycin / Q000494:pharmacology; D016360:Clostridioides difficile / Q000145:classification / Q000187:drug effects / Q000254:growth & development; D003015:Clostridium Infections / Q000382:microbiology*; D003106:Colon / Q000382:microbiology*; D006224:Cricetinae; D004760:Enterocolitis / Q000382:microbiology; D005243:Feces / Q000382:microbiology; D008297:Male; D008647:Mesocricetus; D064307:Microbiota; D008954:Models, Biological
chemical_list: D000900:Anti-Bacterial Agents; D002981:Clindamycin