Cryptococcus neoformans Virulence Assay Using a Galleria mellonella Larvae Model System

Abstract

<i>Cryptococcus neoformans</i> is a human pathogenic fungus that can cause pulmonary infections and meningitis in both immunocompromised and otherwise healthy individuals. Limited treatment options and a high mortality rate underlie the necessity for extensive research of the virulence of <i>C. neoformans</i> . Here we describe a detailed protocol for using the <i>Galleria mellonella</i> (Greater Wax Moth) larvae as a model organism for the virulence analysis of the cryptococcal infections. This protocol describes in detail the evaluation of <i>G. mellonella</i> larvae viability and the alternatives for troubleshooting the infection procedure. This protocol can be easily modified to study different inocula or fungal species, or the effects of a drug or antifungal agent on fungal disease within the larvae. We describe modified alternative versions of the protocol that allow using <i>G. mellonella</i> to study fungal diseases with different inocula and at different temperatures.