Best practices for germ-free derivation and gnotobiotic zebrafish husbandry.

Journal Article (Journal Article;Review)

All animals are ecosystems with resident microbial communities, referred to as microbiota, which play profound roles in host development, physiology, and evolution. Enabled by new DNA sequencing technologies, there is a burgeoning interest in animal-microbiota interactions, but dissecting the specific impacts of microbes on their hosts is experimentally challenging. Gnotobiology, the study of biological systems in which all members are known, enables precise experimental analysis of the necessity and sufficiency of microbes in animal biology by deriving animals germ-free (GF) and inoculating them with defined microbial lineages. Mammalian host models have long dominated gnotobiology, but we have recently adapted gnotobiotic approaches to the zebrafish (Danio rerio), an important aquatic model. Zebrafish offer several experimental attributes that enable rapid, large-scale gnotobiotic experimentation with high replication rates and exquisite optical resolution. Here we describe detailed protocols for three procedures that form the foundation of zebrafish gnotobiology: derivation of GF embryos, microbial association of GF animals, and long-term, GF husbandry. Our aim is to provide sufficient guidance in zebrafish gnotobiotic methodology to expand and enrich this exciting field of research.

Full Text

Duke Authors

Cited Authors

  • Melancon, E; Gomez De La Torre Canny, S; Sichel, S; Kelly, M; Wiles, TJ; Rawls, JF; Eisen, JS; Guillemin, K

Published Date

  • 2017

Published In

Volume / Issue

  • 138 /

Start / End Page

  • 61 - 100

PubMed ID

  • 28129860

Pubmed Central ID

  • PMC5568843

International Standard Serial Number (ISSN)

  • 0091-679X

Digital Object Identifier (DOI)

  • 10.1016/bs.mcb.2016.11.005


  • eng

Conference Location

  • United States