Alternative Non-Mammalian Animal and Cellular Methods for the Study of Host–Fungal Interactions

Abstract

In the study of fungal pathogenesis, alternative methods have gained prominence due to recent global legislation restricting the use of mammalian animals in research. The principle of the 3 Rs (replacement, reduction, and refinement) is integrated into regulations and guidelines governing animal experimentation in nearly all countries. This principle advocates substituting vertebrate animals with other invertebrate organisms, embryos, microorganisms, or cell cultures. This review addresses host–fungus interactions by employing three-dimensional (3D) cultures, which offer more faithful replication of the in vivo environment, and by utilizing alternative animal models to replace traditional mammals. Among these alternative models, species like Caenorhabditis elegans and Danio rerio share approximately 75% of their genes with humans. Furthermore, models such as Galleria mellonella and Tenebrio molitor demonstrate similarities in their innate immune systems as well as anatomical and physiological barriers, resembling those found in mammalian organisms.

Alternative in-vivo models of mucormycosis

“…While C. elegans has been previously employed as a model for studying fungal infections, as comprehensively reviewed by Arvanitis et al. (2013) ; Fusco-Almeida et al. (2023) ; Singulani et al.…”

“…While C. elegans has been previously employed as a model for studying fungal infections, as comprehensively reviewed by Arvanitis et al (2013);Fusco-Almeida et al (2023); Singulani et al (2018); Jacobsen (2019) noted its absence in studying mucormycosis. Recent publications from the group of Vıćtor Meza-Carmen at Universidad Michoacana de San Nicolaś de Hidalgo adopted the model for investigating the virulence of Mucor circinelloides (now classified as M. lusitanicus) knockout strains as well as the toxicity of secreted metabolites in their respective cellfree supernatants.…”

“…Notably, this model can be maintained at temperatures up to 37°C. Fusco-Almeida et al offer a comprehensive review of its applications for medically relevant fungi, however, as of yet, it has not been applied for the study of mucormycosis (Fusco-Almeida et al, 2023).…”