title: Development and validation of a chemostat gut model to study both planktonic and biofilm modes of growth of Clostridium difficile and human microbiota.
abstract: The human gastrointestinal tract harbours a complex microbial community which exist in planktonic and sessile form. The degree to which composition and function of faecal and mucosal microbiota differ remains unclear. We describe the development and characterisation of an in vitro human gut model, which can be used to facilitate the formation and longitudinal analysis of mature mixed species biofilms. This enables the investigation of the role of biofilms in Clostridium difficile infection (CDI). A well established and validated human gut model of simulated CDI was adapted to incorporate glass rods that create a solid-gaseous-liquid interface for biofilm formation. The continuous chemostat model was inoculated with a pooled human faecal emulsion and controlled to mimic colonic conditions in vivo. Planktonic and sessile bacterial populations were enumerated for up to 46 days. Biofilm consistently formed macroscopic structures on all glass rods over extended periods of time, providing a framework to sample and analyse biofilm structures independently. Whilst variation in biofilm biomass is evident between rods, populations of sessile bacterial groups (log10 cfu/g of biofilm) remain relatively consistent between rods at each sampling point. All bacterial groups enumerated within the planktonic communities were also present within biofilm structures. The planktonic mode of growth of C. difficile and gut microbiota closely reflected observations within the original gut model. However, distinct differences were observed in the behaviour of sessile and planktonic C. difficile populations, with C. difficile spores preferentially persisting within biofilm structures. The redesigned biofilm chemostat model has been validated for reproducible and consistent formation of mixed species intestinal biofilms. This model can be utilised for the analysis of sessile mixed species communities longitudinally, potentially providing information of the role of biofilms in CDI.
authors: Leeds Institute of Biomedical and Clinical Sciences, Faculty of Medicine and Health, University of Leeds, Leeds, United Kingdom.
authors: Grace S
authors: GS
authors: Crowther
authors: Leeds Institute of Biomedical and Clinical Sciences, Faculty of Medicine and Health, University of Leeds, Leeds, United Kingdom.
authors: Caroline H
authors: CH
authors: Chilton
authors: Leeds Institute of Biomedical and Clinical Sciences, Faculty of Medicine and Health, University of Leeds, Leeds, United Kingdom.
authors: Sharie L
authors: SL
authors: Todhunter
authors: School of Science, University of West Scotland, Hamilton, United Kingdom.
authors: Scott
authors: S
authors: Nicholson
authors: Department of Microbiology, Leeds Teaching Hospitals NHS Trust, Leeds, United Kingdom.
authors: Jane
authors: J
authors: Freeman
authors: School of Life and Medical Sciences, Department of Human and Environmental Sciences, University of Hertfordshire, Hatfield, United Kingdom.
authors: Simon D
authors: SD
authors: Baines
authors: Leeds Institute of Biomedical and Clinical Sciences, Faculty of Medicine and Health, University of Leeds, Leeds, United Kingdom ; Department of Microbiology, Leeds Teaching Hospitals NHS Trust, Leeds, United Kingdom.
authors: Mark H
authors: MH
authors: Wilcox
mesh_terms: D018441:Biofilms / Q000187:drug effects / Q000254:growth & development*; D016360:Clostridioides difficile / Q000187:drug effects / Q000254:growth & development* / Q000302:isolation & purification; D003470:Culture Media / Q000494:pharmacology; D041981:Gastrointestinal Tract / Q000187:drug effects / Q000382:microbiology*; D006801:Humans; D064307:Microbiota / Q000187:drug effects; D008954:Models, Biological; D010933:Plankton / Q000187:drug effects / Q000254:growth & development*; D015203:Reproducibility of Results
chemical_list: D003470:Culture Media