PUBLICATION

Development of a Larval Zebrafish Infection Model for Clostridioides difficile

Authors
Li, J., Ünal, C.M., Namikawa, K., Steinert, M., Köster, R.W.
ID
ZDB-PUB-200304-8
Date
2020
Source
Journal of visualized experiments : JoVE   (156): (Journal)
Registered Authors
Köster, Reinhard W., Namikawa, Kazuhiko
Keywords
none
MeSH Terms
  • Animals
  • Clostridioides difficile/growth & development
  • Clostridioides difficile/physiology*
  • Clostridium Infections/microbiology
  • Disease Models, Animal
  • Fluorescent Dyes/chemistry
  • Germ-Free Life
  • Intestines/microbiology
  • Intestines/pathology
  • Larva/microbiology
  • Spores, Bacterial/physiology
  • Zebrafish/microbiology*
PubMed
32116293 Full text @ J. Vis. Exp.
Abstract
Clostridioides difficile infection (CDI) is considered to be one of the most common healthcare-associated gastrointestinal infections in the United States. The innate immune response against C. difficile has been described, but the exact roles of neutrophils and macrophages in CDI are less understood. In the current study, Danio rerio (zebrafish) larvae are used to establish a C. difficile infection model for imaging the behavior and cooperation of these innate immune cells in vivo. To monitor C. difficile, a labeling protocol using a fluorescent dye has been established. A localized infection is achieved by microinjecting labeled C. difficile, which actively grows in the zebrafish intestinal tract and mimics the intestinal epithelial damage in CDI. However, this direct infection protocol is invasive and causes microscopic wounds, which can affect experimental results. Hence, a more noninvasive microgavage protocol is described here. The method involves delivery of C. difficile cells directly into the intestine of zebrafish larvae by intubation through the open mouth. This infection method closely mimics the natural infection route of C. difficile.
Genes / Markers
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Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping