SUMMARY: A significant increase in the incidence of Candida -mediated infections has been observed in the last decade, mainly due to rising numbers of susceptible individuals. Recently, the World Health Organization published its first fungal pathogen priority list, with Candida species listed in medium, high, and critical priority categories. This review is a synthesis of information and recent advances in our understanding of two of these species —Candida albicans and Candida glabrata . Of these, C. albicans is the most common cause of candidemia around the world and is categorized as a critical priority pathogen. C. glabrata is considered a high-priority pathogen and has become an increasingly important cause of candidemia in recent years. It is now the second most common causative agent of candidemia in many geographical regions. Despite their differences and phylogenetic divergence, they are successful as pathogens and commensals of humans. Both species can cause a broad variety of infections, ranging from superficial to potentially lethal systemic infections. While they share similarities in certain infection strategies, including tissue adhesion and invasion, they differ significantly in key aspects of their biology, interaction with immune cells, host damage strategies, and metabolic adaptations. Here we provide insights on key aspects of their biology, epidemiology, commensal and pathogenic lifestyles, interactions with the immune system, and antifungal resistance.

Early detection is critical to the successful treatment of life-threatening infections caused by fungal pathogens, as late diagnosis of systemic infection almost always equates with a poor prognosis. The field of fungal diagnostics has some tests that are relatively simple, rapid to perform and are potentially suitable at the point of care. However, there are also more complex high-technology methodologies that offer new opportunities regarding the scale and precision of fungal diagnosis, but may be more limited in their portability and affordability. Future developments in this field are increasingly incorporating new technologies provided by the use of new format biosensors. This overview provides a critical review of current fungal diagnostics and the development of new biophysical technologies that are being applied for selective new sensitive fungal biosensors to augment traditional diagnostic methodologies.

• The amyloid inhibitor Thioflavin-T inhibited C. auris aggregation. • Aggregating isolates do not exhibit any defects in cell separation. • Genomic differences were identified between strongly aggregating and weakly-aggregating strains of C. auris. • Aggregation did not correlate with surface charge or hydrophobicity of yeast cells.

Pathogenic fungi represent an increasing infectious disease threat to humans, especially with an increasing challenge of antifungal drug resistance. Over the decades, numerous tools have been developed to expedite the study of pathogenicity, initiation of disease, drug resistance and host-pathogen interactions. In this review, we highlight advances that have been made in the use of molecular tools using CRISPR technologies, RNA interference and transposon targeted mutagenesis. We also discuss the use of animal models in modelling disease of human fungal pathogens, focusing on zebrafish, the silkworm, Galleria mellonella and the murine model.

Candida auris is a multi-drug resistant human fungal pathogen that has become a global threat to human health due to its drug resistant phenotype, persistence in the hospital environment and propensity for patient to patient spread. Isolates display variable aggregation that may affect the relative virulence of strains. Therefore, dissection of this phenotype has gained substantial interest in recent years. We studied eight clinical isolates from four different clades (I-IV); four of which had a strongly aggregating phenotype and four of which did not. Genome analysis identified polymorphisms associated with loss of cell surface proteins were enriched in weakly-aggregating strains. Additionally, we identified down-regulation of chitin synthase and chitinase genes involved in the synthesis and dissolution of the chitinous septum. Characterisation of the cells revealed no ultrastructural defects in cytokinesis or cell separation in aggregating isolates. Strongly and weakly aggregating strains did not differ in net surface charge or in cell surface hydrophobicity. The capacity for aggregation and for adhesion to polystyrene microspheres were also not correlated. However, aggregation and extracellular matrix formation were all increased at higher growth temperatures, and treatment with the amyloid protein inhibitor Thioflavin-T markedly attenuated aggregation. Genome analysis further indicated strain specific differences in the genome content of GPI-anchored proteins including those encoding genes with the potential to form amyloid proteins. Collectively our data suggests that aggregation is a complex strain and temperature dependent phenomenon that may be linked in part to the ability to form extracellular matrix and cell surface amyloids. HIGHLIGHTS: The amyloid inhibitor Thioflavin-T inhibited C. auris aggregation. Aggregating isolates do not exhibit any defects in cell separation. Genomic differences were identified between strongly aggregating and weakly-aggregating strains of C. auris . Aggregation did not correlate with surface charge or hydrophobicity of yeast cells.

Candida albicans is an opportunistic fungal pathogen that can cause a variety of superficial and life-threatening systemic infections. Relatively few clinically effective antifungal therapies are available, and the increasing prevalence of antifungal drug resistance poses a serious threat in treating these infections. Target validation of biochemical pathways that are essential for fungal growth offers an approach towards the design of novel antifungal drugs that address the growing requirement for new antifungal therapies. Therefore, we used the GRACE library of conditional mutants of C. albicans to explore enzymes in the sugar nucleotide biosynthesis pathway as potential drug targets. This pathway provides UDP-N-acetylglucosamine (UDP-GlcNAc), UDP-glucose (UDP-Glc) and GDP-mannose (GDP-Man) substrates for the synthesis of the essential cell wall polymers, chitin, β-glucan(s) and mannan(s). We show that the genes encoding GDP-mannose pyrophosphorylase (SRB1/PSA1/VIG9), UTP-glucose-1-phosphaturidyl transferase (UGP1), phosphoglucose isomerase (PGI1) and glucosamine-6-phosphate synthase (GFA1) are critical for growth, biofilm formation and virulence in C. albicans. Genes encoding other enzymes in the sugar nucleotide biosynthetic pathway (namely AGM1, PMM1, PMI1, GNA1 and UAP1) were not essential for growth but were required for biofilm formation, tissue invasion and virulence. Repression of genes that encode these enzymes also resulted in hypersensitivity to a range of antifungal drugs as well as oxidative and cell wall stressors. These data underline the potential for augmenting antifungal drug development by targeting these enzymes in the treatment of C. albicans infections.