title: Rearing the Fruit Fly Drosophila melanogaster Under Axenic and Gnotobiotic Conditions.
abstract: The influence of microbes on myriad animal traits and behaviors has been increasingly recognized in recent years. The fruit fly Drosophila melanogaster is a model for understanding microbial interactions with animal hosts, facilitated by approaches to rear large sample sizes of Drosophila under microorganism-free (axenic) conditions, or with defined microbial communities (gnotobiotic). This work outlines a method for collection of Drosophila embryos, hypochlorite dechorionation and sterilization, and transfer to sterile diet. Sterilized embryos are transferred to sterile diet in 50 ml centrifuge tubes, and developing larvae and adults remain free of any exogenous microbes until the vials are opened. Alternatively, flies with a defined microbiota can be reared by inoculating sterile diet and embryos with microbial species of interest. We describe the introduction of 4 bacterial species to establish a representative gnotobiotic microbiota in Drosophila. Finally, we describe approaches for confirming bacterial community composition, including testing if axenic Drosophila remain bacteria-free into adulthood.
authors: Department of Plant and Wildlife Sciences, Brigham Young University.
authors: Melinda L
authors: ML
authors: Koyle
authors: Department of Plant and Wildlife Sciences, Brigham Young University.
authors: Madeline
authors: M
authors: Veloz
authors: Department of Plant and Wildlife Sciences, Brigham Young University.
authors: Alec M
authors: AM
authors: Judd
authors: Department of Entomology, Cornell University; Division of Infectious Diseases, Boston Children's Hospital, Harvard Medical School.
authors: Adam C-N
authors: AC
authors: Wong
authors: Department of Entomology, Cornell University; Biological Sciences, SUNY Oswego.
authors: Peter D
authors: PD
authors: Newell
authors: Department of Entomology, Cornell University; Department of Molecular Biology and Genetics, Cornell University.
authors: Angela E
authors: AE
authors: Douglas
authors: Department of Plant and Wildlife Sciences, Brigham Young University; Department of Entomology, Cornell University; john_chaston@byu.edu.
authors: John M
authors: JM
authors: Chaston
mesh_terms: D000818:Animals; D004330:Drosophila; D004331:Drosophila melanogaster; D041981:Gastrointestinal Tract; D005856:Germ-Free Life; D064307:Microbiota