Search Results (441)

Candidozyma auris (formerly known as Candida auris) has emerged as a formidable clinical fungal pathogen as a result of its multidrug resistance and persistent colonization capabilities. In this study, three clinical C. auris strains (namely C. auris strain 01, C. auris strain 03, and C. auris strain 13) with distinct origins were characterized to investigate their phenotypic variations and mechanisms of azole resistance. Comprehensive profiling revealed significant inter-strain differences in biofilm formation, cell surface hydrophobicity, adhesion capacity, and phospholipase activity. Testing for antifungal susceptibility showed that the three clinical strains exhibited different minimum inhibitory concentrations for multiple azoles (fluconazole, voriconazole, and itraconazole) and echinocandins (anidulafungin and micafungin). Sequencing identified Y132F mutations in the ERG11 gene of the three clinical strains. Mechanistic investigations demonstrated that fluconazole exposure significantly upregulated the expression of efflux pump genes (CDR1 and CDR2) and the genes encoding their transcriptional regulators (MDR1 and TAC1b). In a murine skin colonization model, comparing data from the standard strain C. auris strain CBS12766 and clinical strains of C. auris strain 03 and C. auris strain 13 exhibited a significantly higher fungal burden of tissue, whereas strain C. auris strain 01 showed an intermediate level. Host immunity response analysis revealed that expression of the IL-1β gene was significantly elevated in C. auris strain CBS12766-infected mice, while expression of IL-6 and CXCL-1 genes was predominantly increased in the C. auris strain 01, with TNF-α gene expression levels being comparable across all strains. Histopathological examination confirmed local infiltration of inflammatory cells and mild epidermal edema, indicating active host immune engagement. Overall, our findings highlighted substantial phenotypic heterogeneity, different colonization capacities, and differences in expression of inflammatory cytokines among the C. auris strains. Further investigations into fluconazole-response mechanisms identified enhanced efflux pump activity, along with ERG11 gene Y132F mutations and transcription factor modulation among these clinical strains. Full article

Background: Candidemia remains a significant public health challenge, with increasing resistance. Contemporary post-pandemic data from high-complexity Latin American hospitals are scarce. Methods: A retrospective study (2022–2023) was conducted in adults with candidemia at a high-complexity Colombian university hospital. Species identification and susceptibility were analyzed using VITEK^® 2 and Clinical and Laboratory Standards Institute (CLSI) criteria. Survival was estimated using Kaplan–Meier analysis. Results: Of 3483 blood cultures, 109 episodes were identified. The incidence was 1.13/1000 admissions (5.96/1000 in the Intensive care unit—ICU). Species other than Candida albicans predominated (61.5%), mainly C. tropicalis (22.9%) and C. parapsilosis (22.0%). Alarmingly, 28.6% of C. parapsilosis isolates were resistant to fluconazole. Consultation with an infectious diseases service was performed in 72.5% of cases, with a significantly higher rate among survivors (p < 0.05). Overall mortality was 52.3%, while 30-day mortality reached 42.2%. ICU patients had a cumulative mortality rate of 50% by day 30. Conclusions: Post-pandemic candidemia shows shifting species and high resistance. Key priorities include expert infectious disease consultation to optimize outcomes in non-neutropenic patients and strengthening laboratory capacity for identification and susceptibility testing to monitor rising resistance and guide effective institutional antifungal policies. Full article

Candida auris is a multidrug-resistant fungal pathogen associated with high mortality in healthcare settings. Although colonization is recognized as the harbinger of invasive infection, predicting which patients will develop bloodstream infection (BSI) and when this transition will occur remains a clinical challenge. In this study, patients aged ≥18 years with C. auris colonization identified at İzmir City Hospital between January 2023 and June 2025 were retrospectively analyzed. Colonization was confirmed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Of 71 colonized patients (median age 65 years; 69.0% male; 93.0% intensive care unit (ICU)-admitted), 31 (43.7%) developed bloodstream infection (BSI). In-hospital mortality was 62.0%, rising to 74.2% in the BSI group, though this difference did not reach statistical significance (p = 0.105). Competing risks analysis using the Aalen–Johansen method showed a cumulative BSI incidence of 38.2% (95% confidence interval (CI): 28–50%) by day 10 and 43.0% (95% CI: 32–54%) by day 30 following colonization detection. On multivariate logistic regression, diabetes mellitus was the sole variable independently associated with a lower risk of BSI development (adjusted odds ratio (OR): 0.19; 95% CI: 0.06–0.68; p = 0.010); this finding was directionally consistent but did not reach statistical significance in the multivariable Fine–Gray competing risks model (subdistribution hazard ratio (SHR): 0.334; 95% CI: 0.108–1.040; p = 0.057). All 40 tested isolates had high fluconazole minimum inhibitory concentration (MIC) values; micafungin susceptibility was 92.5%, while anidulafungin resistance was observed in 32.5% of isolates. Our findings demonstrate that nearly half of colonized patients developed BSI, with no identifiable safe window for intervention, underscoring the necessity of sustained infection control measures and susceptibility-guided antifungal therapy. Full article

Invasive candidiasis remains a major cause of morbidity and mortality worldwide, with increasing relevance of non-Candida albicans species, particularly Pichia kudriavzevii, which is associated with high mortality and intrinsic resistance to fluconazole. This study evaluated the effect of voriconazole (VRC) on P. kudriavzevii growth, biofilm formation, and metabolic activity under different nutritional conditions. Planktonic growth and biofilm development were analyzed in Sabouraud dextrose broth (SDB), RPMI-1640, and RPMI-1640 supplemented with glucose (20 g·L⁻¹). Antifungal activity was assessed by optical density (OD₅₇₀) and XTT reduction assays, and biofilm morphology was examined by light microscopy. Glucose consumption was also determined during growth. VRC showed dose-dependent inhibition in SDB, reducing growth and biofilm metabolic activity by up to 94% and 98%, respectively. In contrast, in RPMI-1640, inhibition was significantly lower (≤27% growth and ≤77% biofilm reduction). Glucose supplementation partially restored antifungal susceptibility and increased biofilm metabolic activity. Growth kinetics confirmed VRC-induced delays in proliferation and impaired glucose utilization. These results demonstrate that VRC activity against P. kudriavzevii is strongly dependent on environmental nutrient availability, particularly glucose, which modulates fungal metabolism, biofilm development, and antifungal susceptibility, highlighting the importance of standardized antifungal susceptibility testing conditions and the role of metabolic state in azole efficacy. Full article

Invasive candidiasis caused by drug-resistant Candida species represents a critical global health challenge, with few novel therapeutic scaffolds under development. Here, silver nanoparticles were synthesized using a 21-day fermented Chun Mee kombucha tea extract (K-AgNPs) and characterized by UV-Vis spectroscopy, transmission electron microscopy, nanoparticle tracking analysis, and Fourier-transform infrared spectroscopy. LC-MS/MS profiling of the kombucha substrate documented a phytochemical landscape dominated by epigallocatechin (up to 122,631 µg/mL) and epigallocatechin gallate (up to 415 µg/mL), with a progressive ~80% decline in epicatechin and concomitant increases in gallic acid and chlorogenic acid across the 21-day fermentation. K-AgNPs obtained were spherical, 19.4 nm (±7.9 nm SD) in diameter, with a surface plasmon resonance peak at 415 nm. FTIR confirmed phenolic, carboxylate, and glycosidic surface capping. Antifungal susceptibility testing against eight Candida species, including the WHO critical–priority pathogen Candidozyma auris, showed concordant minimum inhibitory and minimum fungicidal concentrations of 0.80–1.60 µg/mL, confirming fungicidal activity. In vivo evaluation in Galleria mellonella larvae across six infection models demonstrated that K-AgNP treatment at the species-specific MIC significantly improved larval survival versus untreated infected controls (p < 0.01–0.001), while nanoparticle-only groups maintained ≥98% survival, indicating negligible toxicity. Co-treatment amplified total hemocyte mobilization, and K-AgNP-only larvae maintained hemocyte viability above 96% at all time points, indistinguishable from negative controls. Together, these findings demonstrate antifungal activity of K-AgNPs across the genus Candida in standardized in vitro and in vivo settings and provide justification for further investigation, including head-to-head comparison against licensed antifungals and physicochemical validation of nanoparticle stability under assay conditions. Full article

Objectives: When a Candida species is identified in an ICU patient, susceptibility results are typically available in 24–72 h. In this study, we built a machine learning model using four variables available at identification to estimate resistance probability in real time. Methods: We analysed 747 fungal isolates from 725 ICU patients (January 2021–March 2026). We trained and compared a Random Forest and a Logistic Regression model, evaluating both with temporal cross-validation, permutation feature importance, three-category (S/I/R) prediction, and calibration analysis. Results: Multidrug resistance doubled from 24.5% (2021) to 51.1% (2025), and Candida auris grew eight-fold in three years. Random Forest reached AUC 0.885 on the held-out test set and 0.848 on prospective 2024–2025 data (Brier score 0.093). Species identity and drug choice together explained 87% of predictive signal. Local C. albicans fluconazole resistance (~16%) far exceeded the ECMM European figure of 0%, and C. krusei was four times more prevalent than the continental average. Conclusions: A four-variable model may provide calibrated resistance estimates during the critical gap before susceptibility results return, though performance reflects predominantly deterministic species–drug patterns rather than complex learned biology. Overall performance was comparable to a rule-based lookup table, confirming that the majority of predictive signal derives from established species–drug susceptibility patterns. Meaningful added value is limited to temporal trend tracking and improved prediction where resistance is acquired rather than intrinsic (C. albicans, C. tropicalis hard-subset AUC 0.929 vs. rule-based 0.899). The model complements a local antifungal testing; it does not replace one. Full article

The global rise in multidrug-resistant (MDR) fungal pathogens has positioned Candida auris and Candida glabrata as major threats to public health. In recent years, these pathogens have increasingly been reported beyond traditional hospital settings, including neonatal intensive care units, long-term care facilities, oncology wards, and post-pandemic critical care environments. International surveillance bodies, including the Centers for Disease Control and Prevention (CDC), European Centre for Disease Prevention and Control (ECDC), World Health Organization (WHO), and regional monitoring networks, have documented escalating antifungal resistance, complex outbreak dynamics, and persistent gaps in infection control implementation. C. auris has emerged as a major etiological agent of healthcare-associated outbreaks, particularly in intensive care and neonatal units. Surveillance data indicate that a high proportion of C. auris isolates exhibit resistance to azoles, often exceeding 80% in some regions, while echinocandin resistance remains variable. Resistance patterns have evolved from predominantly azole resistance to broader multidrug-resistant phenotypes, including treatment-emergent echinocandin resistance. Six genetically distinct clades (I–VI) have been identified, with Clades I, III, and IV associated with large-scale outbreaks, whereas available data suggests that Clades II, V, and VI are more geographically restricted, although evidence for the recently described clades remains limited. C. glabrata is increasingly recognized as a major cause of invasive candidiasis, with rising resistance reported across multiple regions. While reduced azole susceptibility was historically predominant, emerging evidence highlights rising dual azole–echinocandin resistance, adaptive microevolution during antifungal therapy, and biofilm-associated tolerance mechanisms. Despite these advances, significant gaps persist in global resistance surveillance and in the mechanistic understanding of virulence and antifungal adaptation. Current mitigation strategies include antifungal stewardship programs, expanded resistance testing, and strengthened surveillance systems. Advances in rapid diagnostic technologies such as matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry, polymerase chain reaction (PCR)-based assays, and genomic surveillance have improved pathogen identification and outbreak detection, although accessibility remains limited in resource-constrained settings. This review examines emerging epidemiological, genomic, and antifungal resistance trends in C. auris and C. glabrata and highlights key priorities for improving diagnosis, surveillance, stewardship, and management of multidrug-resistant Candida infections. Full article

Malassezia are commensal lipid dependent yeasts which can cause opportunistic skin infection. Topical imidazole antifungals such as clotrimazole and ketoconazole are the frontline treatment. However, the tendency of fungal infections to recur, combined with the emergence of multi-azole-resistant Malassezia isolates means that many patients have used these antifungal treatments repeatedly or for extended durations with limited efficacy. While the impact of single azole treatments has been studied, the ability of specific azoles to induce cross-resistance is unclear. Understanding the effect of prior exposure of one treatment on susceptibility to other antifungals is important in the selection of the appropriate treatment to avoid driving the evolution of greater resistance. We previously identified drug transporters from the ATP-Binding Cassette (ABC) and Major Facilitator Superfamily (MFS) to be upregulated on extended exposure to clotrimazole. In this study, we investigated the effect of extended clotrimazole, ketoconazole and fluconazole exposure on antifungal cross-resistance profiles and examined the expression of the MFS transporters OPT1 and FLR1 in resistance emergence. We observed that treatment with clotrimazole was associated with increased cross-resistance to other antifungals. Ketoconazole treatment caused elevated MICs in all tested antifungals that did not decrease after drug removal. These findings advance our understanding of fungal adaptive resistance mechanisms and inform improved antifungal strategies to mitigate resistance development. Full article

Aspergillus spp. are common environmental molds worldwide and mostly cause infections in immunocompromised individuals. We have previously shown that black aspergilli (Aspergillus section Nigri) isolates are the most common aspergilli in indoor and outdoor hospital environments in Kuwait. This study reports on the relative prevalence of different Aspergillus section Nigri complex species among clinical isolates and their susceptibility to antifungal drugs. Black aspergilli isolated from clinical (n = 34) and environmental (n = 2) sources were studied. The isolates were initially identified as Aspergillus section Nigri complex members based on morphological characteristics. Species-specific identification was carried out by PCR-sequencing of the β-tubulin gene fragment and sequence comparisons with the GenBank database. The phylogenetic analysis was performed by using the Maximum Likelihood method in MEGA (version 11) software. Antifungal susceptibility testing was performed by Etest. The phylogenetic analysis based on β-tubulin gene sequences identified only three species: A. niger sensu stricto (A. niger) (n = 26), A. tubingensis (n = 7), and A. luchuensis (n = 1) among 34 clinical Aspergillus section Nigri isolates in Kuwait. All seven otomycoses cases were due to A. niger. The two environmental isolates were identified as A. niger and A. tubingensis. All isolates appeared susceptible to all five (amphotericin B, itraconazole, voriconazole, posaconazole, and caspofungin) antifungal drugs tested. In conclusion, our study shows that A. niger predominates among phenotypically identified clinical isolates of the Aspergillus section Nigri complex, and that A. niger is also the main agent in otomycosis cases in Kuwait. The detection of only three species within the Aspergillus section Nigri complex in Kuwait could be due to the limited number (n = 34) of clinical isolates analyzed in this study. Full article

Vulvovaginal candidiasis (VVC) is a common fungal infection among reproductive-age women and is increasingly challenged by the emergence of non-albicans Candida species and reduced azole susceptibility. This prospective cross-sectional study investigated 235 symptomatic reproductive-age women attending two healthcare facilities in Ho Chi Minh City, Vietnam, to determine VVC prevalence, Candida species distribution, pregnancy-associated patterns, antifungal susceptibility, and diagnostic performance. Vaginal swabs were cultured on Sabouraud Dextrose Agar and CHROMagar™ Candida, while species identification was confirmed by PCR-RFLP targeting the ITS region. Susceptibility to fluconazole and clotrimazole was assessed using the disk diffusion method. Candida spp. was detected in 55.7% of participants. C. albicans accounted for 50.3% of isolates, whereas non-albicans Candida species represented 49.7%, indicating a substantial species shift. VVC was more frequent among pregnant women, particularly in the third trimester. Most C. albicans, C. tropicalis, and C. parapsilosis isolates remained susceptible to azoles; however, C. glabrata showed markedly reduced susceptibility to fluconazole and clotrimazole. CHROMagar™ Candida reliably identified C. albicans but misclassified several non-albicans Candida isolates compared with PCR-RFLP. These findings highlight the need for routine species-level diagnosis, antifungal susceptibility testing, and strengthened VVC surveillance in reproductive and antenatal healthcare settings in Vietnam. Full article

Mucormycosis is a devastating invasive fungal infection primarily caused by Mucor and Rhizopus species, presenting significant clinical challenges due to limited therapeutic options and emerging drug resistance in opportunistic yeasts such as Candida albicans. This study explores foliar endophytic fungi from Thai medicinal plants as potential reservoirs for novel bioactive metabolites targeting both fungal growth and virulence factors. We report the first isolation of Fusarium proliferatum as an endophyte from Lantana camara L. foliage (voucher number 01562), with its identity confirmed through morphological characterization and sequencing of the fungal ITS4/ITS5 regions. Antifungal susceptibility testing showed potent activity against a panel of environmental Mucorales, with minimum inhibitory concentrations (MICs) ranging from 0.3 to 1 mg/L. In dual-culture assays, F. proliferatum demonstrated significant mycelial inhibition rates of 93.30% to 93.67% against Mucor spp. and 88.67% to 93.67% against Rhizopus spp. Furthermore, the crude extract exhibited a potent anti-virulence effect by suppressing the C. albicans yeast-to-hyphal transition, achieving up to 68% germination inhibition in resistant strains. Liquid chromatography–mass spectrometry (LC-MS) analysis identified 51 secondary metabolites, including the cyclic peptide beauvericin and various polyketides and indole derivatives. These findings suggest that F. proliferatum utilizes metabolic mimicry and adaptive synergy with its host plant to produce a diverse chemical arsenal. This study positions foliar endophytes of L. camara as promising candidates for the development of dual-action therapeutics to combat invasive and resistant mycoses. Full article

The mitochondrial membrane protein UPS1, a conserved intermembrane space protein in Saccharomyces cerevisiae, possesses phosphatidic acid transfer activity and plays a positive regulatory role in processes such as cardiolipin metabolism and transport. The role of UPS1 protein in pathogenic fungi such as Candida albicans has not been explored, especially in relation to its influence on virulence factors like hyphal growth and biofilm formation, which are crucial for the pathogenicity of C. albicans. The research investigated the function of the UPS1 protein in C. albicans by using gene knockout techniques, analyzing mitochondrial function, and conducting tests for hyphal and biofilm development. The results revealed that deletion of the UPS1 gene leads to altered mitochondrial morphology, increased reactive oxygen species levels, and reduced intracellular ATP content, thereby causing severe growth defects in C. albicans. In addition, transcriptomic analysis indicated that loss of UPS1 significantly represses the expression of genes associated with hyphal growth and biofilm formation. Functional assays further confirmed that UPS1 deficiency markedly impairs cell adhesion capability, hyphal development, and biofilm formation of C. albicans. Notably, deletion of the UPS1 protein markedly reduces the susceptibility of C. albicans to membrane-targeted antifungal drugs. Finally, infection models using Galleria mellonella larvae and a murine vulvovaginal candidiasis model verified that UPS1 gene knockout attenuates the pathogenicity of C. albicans. In summary, our findings demonstrate that UPS1 protein modulates the pathogenicity of C. albicans by regulating mitochondrial function, hyphal growth, and biofilm formation. Full article

Candida albicans remains one of the leading causes of invasive fungal infections and is recognized as a critical-priority pathogen by the World Health Organization. The increasing emergence of resistance to azole antifungals such as fluconazole highlights the need for alternative therapeutic strategies. In this study, we evaluated the antifungal potential of a chromium(III)–triazole coordination complex (CrL1) against C. albicans. In vitro susceptibility testing showed that CrL1 exhibited notable antifungal activity against the fluconazole-resistant strain with low cytotoxicity in murine macrophages. To facilitate aqueous dispersion and enable in vivo administration, CrL1 was incorporated into an oil-in-water nanoemulsion (NE-CrL1). The antifungal activity of NE-CrL1 was evaluated in a murine model of invasive candidiasis. In mice infected with a fluconazole-resistant C. albicans strain, treatment with NE-CrL1 reduced renal fungal burden and was associated with attenuation of histopathological alterations and changes in local inflammatory responses. Although the present study has limitations, including the absence of mechanistic assays and additional physicochemical characterization, these results suggest in vivo antifungal activity of NE-CrL1 and warrant further preclinical evaluation against drug-resistant Candida infections. Full article

Aspergillosis encompasses a heterogeneous spectrum of diseases caused by filamentous fungi of the genus Aspergillus, ranging from allergic airway disorders and chronic pulmonary infection to rapidly progressive invasive disease. Aspergillus fumigatus is the predominant pathogen worldwide, although other species, including Aspergillus flavus, Aspergillus terreus and cryptic species, contribute to morbidity and may exhibit intrinsic or acquired antifungal resistance. Early and accurate laboratory diagnosis is essential for timely treatment, appropriate antifungal selection, and stewardship. Traditional culture remains foundational, enabling confirmation of viable organisms, species-level identification, and antifungal susceptibility testing, but sensitivity is limited and turnaround times are prolonged. Non-culture approaches—including galactomannan, β-D-glucan, lateral flow assays, PCR, and next-generation sequencing—enhance diagnostic sensitivity, facilitate early detection, and allow identification of resistance-associated mutations. Optimal diagnostic performance is achieved through integrated, multimodal strategies combining laboratory tests with clinical and radiological findings. In invasive disease, concurrent use of biomarkers and molecular assays improves specificity and positive predictive value, while in allergic bronchopulmonary aspergillosis, immunological markers remain central. Future directions include standardised molecular protocols, novel antigenic and host-based biomarkers, and cost-effective, risk-adapted diagnostic algorithms to refine detection, guide therapy, and improve patient outcomes. Full article

Background/Objective: The introduction of Ketoconazole (KZ, Nizoral^®) in 1977 by Janssen Pharmaceutica marked a significant milestone in medical mycology as the first broad-spectrum oral antifungal agent. However, KZ is a highly lipophilic compound, presenting significant challenges in the development of efficient topical formulations. Moreover, oral KZ has undergone labeling revisions and market withdrawal due to serious hepatic side effects. This study aimed to design, optimize, and evaluate KZ-loaded nanoemulsions (NEs; KZ-NEs) as a delivery platform that could improve skin bioavailability and antifungal activity. Methods: Optimized KZ-NEs were converted to a mucoadhesive formulation (KZ-NEC) by the addition of Carbopol^® 940 NF to enhance the adherence of the formulations to the skin surface. NEs were evaluated concerning physical appearance, globule size, polydispersity index, zeta potential, pH, viscosity, and drug content. Optimized KZ-NE and lead KZ-NEC formulations were further evaluated for in vitro release, ex vivo skin permeation and deposition, skin irritation, and in vivo studies. Results: In vitro release studies revealed that nanocarrier systems provided a sustained release of KZ over 24 h. The ex vivo permeability coefficients of KZ from the optimized KZ-NE and lead KZ-NEC formulations were approximately four- and three-fold greater than that achieved with the marketed cream formulation, respectively. In addition, the C_max of the lead KZ-NEC formulation (14.4 ± 1.1 μg/mL) was significantly higher (p < 0.05) compared with the marketed cream formulation (10.5 ± 0.5 μg/mL). Moreover, in vitro antifungal susceptibility testing showed that KZ demonstrated improved antifungal efficacy when incorporated into the KZ-NE and KZ-NEC formulations. Neither of the NE-based formulations caused any alterations in skin color or morphology during the 24 h visual observation period. Both NE-based formulations were stable for 90 days (the last time-point tested) at three different storage conditions. Conclusions: NE-based formulation could serve as an effective topical delivery platform for KZ and could improve therapeutic outcomes for patients with topical fungal infections. Full article