A Galleria mellonella Oral Administration Model to Study Commensal-Induced Innate Immune Responses

Our protocol expands the use of G.mellonella from a systemic infection model to a model that allows bacterial administration via the natural oral route. G.mellonella is a suitable replacement model for rodents in analyzing different hallmarks of innate micro-post interactions as the larvae can be used for human pathogenic bacteria. Begin by transferring the eggs laid by adult moths to two liter boxes containing wax moth substrate, at 30 degrees Celsius, in the dark.

After one to two weeks, transfer 25 grams of larva containing substrate into fresh substrate, when small and tiny larva become visible. Synchronize the larvae after two weeks according to their size, into groups of 30 to 40 larvae in new two liter containers on wax moth substrate. After two weeks, select larvae for the experiment by weight.