The use of embryonated egg as an alternative in the study of the pathogenesis of fungi is evolving. Although murine models are the “gold standard,” embryonated egg models are also used to screen determinants of virulence among fungi species. This study was aimed at determining the virulence potential of Cryptococcus gattii strains R265, R272, and EJB18, and Malassezia sympodialis using chorioallantoic membrane (CAM) of embryonated egg. At a concentration of 107 cfu/ml, C. gattii R272 was more virulent than R265 in the egg model, while EJB18 had low virulence. The CAM model supported the growth of Malassezia sympodialis strain and induced the formation of hyphae. The formation of lesions by the organism and its re-isolation from CAM suggest that the model can be used for evaluating the virulence of C. gattii. Histopathology of CAM from both strains also revealed massive disruption of CAM. This study suggests that embryonated egg is a useful alternative tool to pre-screen Cryptococcus gattii strains to select strains for subsequent testing in murine models and could also be a potential medium for studying the hyphal growth in Malassezia species.

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Echinocandins are antifungal agents that specifically inhibit the biosynthesis of 1,3-β-D-glucan, a major structural component of fungal cell walls. Echinocandins are recommended as first-line or alternative/salvage therapy for candidiasis and aspergillosis in antifungal guidelines of various countries. Resistance to echinocandins has been reported in recent years. The mechanism of echinocandin resistance involves amino acid substitutions in hot spot regions of the FKS gene product, the catalytic subunit of 1,3-β-D-glucan synthase. This resistance mechanism contributes to not only acquired resistance in Candida spp., but also inherent resistance in some pathogenic fungi. An understanding of the echinocandin resistance mechanism is important to develop both effective diagnosis and treatment options for echinocandin-resistant fungal diseases.

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　Aspergillus fumigatus is the predominant fungal pathogen responsible for life-threatening systemic infections in humans. Recently developed high-throughput whole genome sequencing (WGS) and RNA-Seq technologies have proven to be powerful tools for systematically investigating pathogenic organisms. In this review, we present new virulence factors obtained through our “omics” researches on A. fumigatus.
　We first sequenced genomes of A. fumigatus stains isolated from one infected patient at different time points, and made an important finding that although the genome (microsatellites) type of the infected strain remained unchanged, the strain exhibited several genetic changes, including acquiring therapeutic drug resistance, during patient treatment for 1.5 years.
　Of the various presentations of aspergillosis, pulmonary aspergilloma (PA) is one of the most common forms of A. fumigatus infection, where fungus balls are composed of fungal hyphae, inflammatory cells, fibrin, mucus, and tissue debris. Chronic necrotizing pulmonary aspergillosis (CNPA), also known as semi-invasive or invasive aspergillosis, is locally invasive and predominantly seen in patients with mild immunodeficiency or with a chronic lung disease. We compared genomes of strains individually isolated from eight PA and eight CNPA patients in Japan, and found that the PA and CNPA strains show indiscernible genetic and ancestral backgrounds as far as genomic SNPs of the strains are concerned.
　The main route of infections caused by A. fumigatus is via inhalation of conidia. Inhaled conidia rapidly adhere to pulmonary epithelial cells. Nevertheless, little is known of the molecular mechanism of adherence in A. fumigatus resting conidia. We assumed corresponding adhesion molecules were highly expressed in high-adhesion conidia during conidia maturation, and exhaustively searched adhesion molecules by comparing gene expression levels in high- and low-adherence strains using the RNA-Seq technique. We found several factors involved in conidial adhesion and suggest that composite actions of these molecules have roles in conidial adhesion to human pulmonary epithelial cells.

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