Rearing and Maintenance of Galleria mellonella and Its Application to Study Fungal Virulence

Carolina Firacative,Shuyao Duan,Kennio Ferreira-Paim,Diana Leemon,Wieland Meyer,Aziza Khan

doi:10.3390/jof6030130

Abstract

Galleria mellonella larvae have been widely used as alternative non-mammalian models for the study of fungal virulence and pathogenesis. The larvae can be acquired in small volumes from worm farms, pet stores, or other independent suppliers commonly found in the United States and parts of Europe. However, in countries with no or limited commercial availability, the process of shipping these larvae can cause them stress, resulting in decreased or altered immunity. Furthermore, the conditions used to rear these larvae including diet, humidity, temperature, and maintenance procedures vary among the suppliers. Variation in these factors can affect the response of G. mellonella larvae to infection, thereby decreasing the reproducibility of fungal virulence experiments. There is a critical need for standardized procedures and incubation conditions for rearing G. mellonella to produce quality, unstressed larvae with the least genetic variability. In order to standardize these procedures, cost-effective protocols for the propagation and maintenance of G. mellonella larvae using an artificial diet, which has been successfully used in our own laboratory, requiring minimal equipment and expertise, are herein described. Examples for the application of this model in fungal pathogenicity and gene knockout studies as feasible alternatives for traditionally used animal models are also provided.

Highlights

The greater wax moth Galleria mellonella (Lepidoptera: Pyralidae) is a ubiquitous pest of honeybee colonies globally causing damage to wax combs in stressed beehives and stored beekeeping equipment, where the larvae feed and transform into moths [1]
Results from the murine experiments were obtained from our previous study [14]
To show the capacity of the G. mellonella larvae model to study fungal virulence, the larvae were inoculated with the high-virulence C. gattii strain CDCR265, which began dying 96 h after inoculation, and with the cell capsules red.with

Summary

Introduction

The greater wax moth Galleria mellonella (Lepidoptera: Pyralidae) is a ubiquitous pest of honeybee colonies globally causing damage to wax combs in stressed beehives and stored beekeeping equipment, where the larvae feed and transform into moths [1]. G. mellonella can undergo a complete life cycle in 8 to 12 weeks under favorable conditions of temperature and humidity [1]. Eggs hatch to larvae, which undergo 7 moults (instars) before pupation and metamorphosis into adults. The cream-colored 6th instar larvae, which are about 3 cm in length and weigh approximately. 300 mg, have become increasingly popular as an alternative non-mammalian model to study fungal disease and virulence [2,3,4,5,6,7]. G. mellonella larvae have several advantages including.

Results

Discussion

Conclusion