Candida auris is an emerging multidrug-resistant yeast that causes healthcare-associated and deep-seated infections. Notably, the emergence of this yeast is alarming as it exhibits resistance to azoles, echinocandins, and amphotericin B, which may lead to clinical treatment failure in patients. This study aims to identify and characterize the genetic determinants of antifungal resistance in C. auris among some local clinical isolates to contribute for understanding the molecular epidemiology of C. auris in Egypt. Four test strains were identified based on the ribosomal internal transcribed spacer (ITS) region sequence and phylogenetic analysis. Antifungal susceptibility was determined using the VITEK 2 system. Molecular analysis of ERG11, ERG3, FKS1, and FKS2 was used to identify mutations associated with antifungal resistance. The four test strains were identified as C. auris. Evolutionary analysis was conducted, and sequences of ITS regions were submitted to GenBank. The mutations Y132F in ERG11 and F635Y in FKS2 were identified, which are known to confer resistance to azoles and echinocandins, respectively. The emergence of C. auris in Egypt represents a public health concern. Hospitals should implement strict infection control measures to prevent its spread. Effective treatment guidelines and ongoing monitoring of antifungal resistance are essential to combat this emerging pathogen.

The fungal secretome is the main interface for interactions between the pathogen and its host. It includes the most important virulence factors and effector proteins. We integrated different bioinformatic approaches and used the newly drafted genome data of P. lingam isolate CAN1 (blackleg of rapeseed fungus) to predict the secretion of 217 proteins, including many cell-wall-degrading enzymes. All secretory proteins were identified; 85 were classified as CAZyme families and 25 were classified as protease families. Moreover, 49 putative effectors were predicted and identified, where 39 of them possessed at least one conserved domain. Some pectin-degrading enzymes were noticeable as a clustering group according to STRING web analysis. The secretome of P. lingam CAN1 was compared to the other two blackleg fungal species (P. lingam JN3 and P. biglobosus CA1) secretomes and their CAZymes and effectors were identified. Orthologue analysis found that P. lingam CAN1 shared 14 CAZy effectors with other related species. The Pathogen-Host Interaction database (PHI base) classified the effector proteins in several categories where most proteins were assigned as reduced virulence and two of them termed as hypervirulence. Nowadays, in silico approaches can solve many ambiguous issues about the mechanism of pathogenicity between fungi and plant host with well-designed bioinformatics tools.

Candidozyma auris (formerly Candida auris) has emerged as a significant multidrug-resistant pathogen. Among 140 antifungal-resistant Candida spp. isolates, 120 were identified as C. auris using chromogenic agar, VITEK 2, and MALDI-TOF. Most cases (60%) were males, and 59% were aged ≥ 60 years. Bloodstream infections were predominant (60.8%), followed by respiratory (20.8%), urinary tract (15%), and other sites. Liver transplantation (35.8%) was the most common underlying risk factor, followed by post-COVID-19 (30.8%) and cancer (25%). All isolates exhibited thermotolerance, halotolerance, anaerobic blood hemolysis, and biofilm formation. Significant association was observed between virulence enzymes activity and both specimen source and clinical conditions (P < 0.001), with strong activity linked to blood isolates, liver transplant, post-COVID-19, and lung cancer. Among antifungal agents, voriconazole (MIC₅₀/₉₀: 0.125/0.5 µg/mL), posaconazole (0.03/0.125 µg/mL), and amphotericin B (0.5/1 µg/mL) showed consistently low MICs. Caspofungin (0.25/1 µg/mL) and micafungin (0.125/0.5 µg/mL) demonstrated favorable activity, with resistance rates of 3.3% and 4.2%, respectively. Fluconazole resistance was observed among all isolates (MIC₅₀/₉₀: 32/32 µg/mL). Itraconazole and ketoconazole showed elevated MICs (MIC₅₀/₉₀: 1/4 and 16/64 µg/mL, respectively) and was inactive against all isolates. Flucytosine (MIC₅₀/₉₀: 64/128 µg/mL) was inactive against 96.7%. Agreement between VITEK 2 and CLSI BMD was strong for fluconazole, voriconazole, amphotericin B, caspofungin, and micafungin (Cohen’s kappa ≥ 0.8), but poor for flucytosine (0.35), indicating limited reliability. This report provides crucial local data on antifungal susceptibility and virulence traits of C. auris, supporting infection control and treatment strategies in Egypt.

Covid-19 crisis did hit many socio-economic aspects in the whole world. In the scientific research, the problem is getting even worse, since most of materials and consumable are allocated to the health sector. Many research laboratories around the world have big delay in receiving their purchases to accomplish their research projects. In the developing countries, the situation is much more difficult, since most of the funding resources are directed to the Covid-19 crisis and there is a notable increase in reagents’ prices. Therefore, the aim of the present study is to make a homemade reagents for RNA purification from eukaryotic cells/tissues. The homemade phenol-based RNA extraction reagents were prepared using saturated phenol pH 4.3 (adjusted by 0.5 M citrate buffer) and guanidine thiocyanate. To validate the phenol-based reagent, RNA was purified from different biological samples (cell line, tissues, and fungi) using homemade phenol-based versus a commercial one. Concentration of RNA samples extracted from the same number of cells were compared to assess the homemade preparation of phenol-based reagent. In conclusion, homemade phenol-based reagent is cost effective and comparable to the commercial one. Using homemade phenol-based, RNA extraction was successfully purified from different biological sources.