Infection of Galleria mellonella (Lepidoptera) Larvae With the Entomopathogenic Fungus Conidiobolus coronatus (Entomophthorales) Induces Apoptosis of Hemocytes and Affects the Concentration of Eicosanoids in the Hemolymph

Apoptosis and autophagy, the mechanisms of programmed cell death, play critical roles in physiological and pathological processes in both vertebrates and invertebrates. Apoptosis is also known to play an important role in the immune response, particularly in the context of entomopathogenic infection. Of the factors influencing the apoptotic process during infection, two of the lesser known groups are caspases and eicosanoids. The aim of this study was to determine whether infection by the entomopathogenic soil fungus Conidiobolus coronatus is associated with apoptosis and changes in caspase activity in the hemocytes of Galleria mellonella larvae, and to confirm whether fungal infection may affect eicosanoid levels in the host. Larvae were exposed for 24 h to fully grown and sporulating fungus. Hemolymph was collected either immediately after termination of exposure (F24 group) or 24 h later (F48 group). Apoptosis/necrosis tests were performed in hemocytes using fluorescence microscopy and flow cytometry, while ELISA tests were used to measure eicosanoid levels. Apoptosis and necrosis occurred to the same degree in F24, but necrosis predominated in F48. Fungal infection resulted in caspase activation, increased PGE1, PGE2, PGA1, PGF2α, and 8-iso-PGF2α levels and decreased TXB2 levels, but had no effect on TXA2 or 11-dehydro-TXB2 concentrations. In addition, infected larvae demonstrated significantly increased PLA2 activity, known to be involved in eicosanoid biosynthesis. Our findings indicate that fungal infection simultaneously induces apoptosis in insects and stimulates general caspase activity, and this may be correlated with changes in the concentrations of eicosanoids.

The Impact of the Entomopathogenic Fungus Conidiobolus coronatus on the Free Fatty Acid Profile of the Flesh Fly Sarcophaga argyrostoma

The chemical composition of the insect cuticle varies remarkably between species and their life stages. It can affect host resistance and substrate utilization by invading entomopathogen fungi, such as the soil fungus Conidiobolus coronatus. In this study, Sarcophaga argyrostoma flies were exposed to sporulating C. coronatus colonies for 24 h; the pupae were resistant, but the adults demonstrated 60% mortality. Although the pupae demonstrated no sign of infection nor any abnormal development, our findings indicate that after 24 h of contact with the fungus, the pupae demonstrated a 25.2-fold increase in total cuticular free fatty acids (FFAs) and a 1.9-fold decrease in total internal FFAs. Also, the cuticular FFA increased from 26 to 30, while the internal FFA class increased from 13 to 23. In exposed adults, the total mass of cuticular FFAs increased 1.7-fold, while the number of FFAs stayed the same (32 FFAs). Also, the internal FFA class increased from 26 to 35 and the total FFA mass increased 1.1-fold. These considerable differences between adults and pupae associated with C. coronatus exposure indicate developmental changes in the mechanisms governing lipid metabolism and spatial distribution in the organism, and suggest that cuticular lipids play a vital role in the defence against pathogenic fungi.

Dodecanol, metabolite of entomopathogenic fungus Conidiobolus coronatus, affects fatty acid composition and cellular immunity of Galleria mellonella and Calliphora vicina

One group of promising pest control agents are the entomopathogenic fungi; one such example is Conidiobolus coronatus, which produces a range of metabolites. Our present findings reveal for the first time that C. coronatus also produces dodecanol, a compound widely used to make surfactants and pharmaceuticals, and enhance flavors in food. The main aim of the study was to determine the influence of dodecanol on insect defense systems, i.e. cuticular lipid composition and the condition of insect immunocompetent cells; hence, its effect was examined in detail on two species differing in susceptibility to fungal infection: Galleria mellonella and Calliphora vicina. Dodecanol treatment elicited significant quantitative and qualitative differences in cuticular free fatty acid (FFA) profiles between the species, based on gas chromatography analysis with mass spectrometry (GC/MS), and had a negative effect on G. mellonella and C. vicina hemocytes and a Sf9 cell line in vitro: after 48 h, almost all the cells were completely disintegrated. The metabolite had a negative effect on the insect defense system, suggesting that it could play an important role during C. coronatus infection. Its high insecticidal activity and lack of toxicity towards vertebrates suggest it could be an effective insecticide.

Amino Acids in Entomopathogenic Fungi Cultured In Vitro

The content of bounded amino acids in six entomopathogenic fungi was identified and determined. Analyzing the elements characterizing the pathogenicity of individual species of fungi based on infectivity criteria, ranges of infected hosts, and the ability to induce epizootics, these can be ranked in the following order: Isaria farinosa, Isaria tenuipes, Isaria fumosorose, Lecanicillium lecanii, Conidiobolus coronatus, Isaria coleopterorum. These fungi represent two types of Hyphomycetales-Paecilomyces Bainier and Verticillium Nees ex Fr. and one type of Entomophtorales-Conidiobolus Brefeld. Our study indicates that there are significant quantitative and qualitative differences of bounded amino acids in the entomopathogenic fungal strains contained in the mycelium between high and low pathogenicity strains. The richest composition of bounded amino acids has been shown in the mycelium of the Isaria farinosa strain, which is one of the most commonly presented pathogenic fungi in this group with a very wide range of infected hosts and is the most frequently recorded in nature as an important factor limiting the population of insects.

Rhinofacial Conidiobolomycosis: A Case Series and Review of the Literature

Conidiobolomycosis is an uncommon, chronic, localized subcutaneous mycosis primarily affecting rhinofacial region. It is reported mainly from tropical and subtropical countries. The condition is underreported due to the lack of clinical suspicion and usually mismanaged. This rare mycosis is due to the genus Conidiobolus within the order Entomophthorales of class Zygomycetes. Here we present 3 cases of rhinofacial conidiobolomycosis in otherwise healthy adults from different parts of Sri Lanka over 1-year period. All patients had disfiguring subcutaneous lesions in the rhinofacial area. The diagnoses were based on isolation of Conidiobolus coronatus in clinical specimens.