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Rearing zombie flies: Laboratory culturing of the behaviourally manipulating fungal pathogen Entomophthora muscae .

MethodsX 2024 June
Insect pathogenic fungi (IPF) and insects have ubiquitous interactions in nature. The extent of these interkingdom host-pathogen interactions are both complex and diverse. Some IPF, notably of the order Entomophthorales, manipulate their species-specific host before death. The fungus-induced altered insect behaviours are sequential and can accurately be repeatedly characterised temporally, making them a valuable model for understanding the molecular and chemical underpinnings of behaviour and host-pathogen co-evolutionary biology. Here, we present methods for the isolation and laboratory culturing of the emerging behaviourally manipulating model IPF Entomophthora muscae for experimentation.• E. muscae isolation and culturing in vitro .•Establishing and maintaining an E. muscae culture in vivo in houseflies ( Musca domestica ).•Controlled E. muscae infections for virulence experiments and quantification of conidia discharge per cadaver.

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