Overview of rearing and testing conditions and a guide for optimizing Galleria mellonella breeding and use in the laboratory for scientific purposes.

The greater wax moth Galleria mellonella is an increasingly popular and consolidated alternative infection model to assess microbial virulence and the effectiveness of antimicrobial compounds. The lack of G. mellonella suppliers aiming at scientific purposes and a lack of well-established protocols for raising and testing these animals may impact results and reproducibility between different laboratories. In this review, we discuss the state of the art of rearing the larvae in situ, providing an overview of breeding and testing conditions commonly used and their influence on larval health and experiments results, from setting up the environment, providing the ideal diet, understanding the effects of pretreatments, choosing the best testing conditions, to exploring the most from the results obtained. Meanwhile, we guide the reader through the most practical ways of dealing with G. mellonella to achieve successful experiments.