The genus Aspergillus harbors human infection-causing pathogens and is involved in the complex one-health challenge of antifungal resistance. Here, a 6-year retrospective study was conducted with Aspergillus spp. isolated from patients with invasive, chronic, and clinically suspected aspergillosis in a tertiary teaching hospital. A total of 64 Aspergillus spp. clinical isolates were investigated regarding molecular identification, biofilm, virulence in Galleria mellonella, antifungal susceptibility, and resistance to amphotericin B and azoles. Aspergillus section Fumigati (A. fumigatus sensu stricto, 62.5%) and section Flavi (A. flavus, 20.3%; A. parasiticus, 14%; and A. tamarii, 3.1%) have been identified. Aspergillus section Flavi clinical isolates were more virulent than section Fumigati clinical isolates. Furthermore, scant evidence supports a link between biofilm formation and virulence. The susceptibility of the Aspergillus spp. clinical isolates to itraconazole, posaconazole, voriconazole, and amphotericin B was evaluated. Most Aspergillus spp. clinical isolates (67.2%) had an AMB MIC value equal to or above 2 µg/mL, warning of a higher probability of therapeutic failure in the region under study. In general, the triazoles presented MIC values above the epidemiological cutoff value. The high triazole MIC values of A. fumigatus s.s. clinical isolates were investigated by sequencing the promoter region and cyp51A locus. The Cyp51A amino acid substitutions F46Y, M172V, N248T, N248K, D255E, and E427K were globally detected in 47.5% of A. fumigatus s.s. clinical isolates, and most of them are associated with high triazole MICs. Even so, the findings support voriconazole or itraconazole as the first therapeutic choice for treating Aspergillus infections. This study emphasizes the significance of continued surveillance of Aspergillus spp. infections to help overcome the gap in knowledge of the global fungal burden of infections and antifungal resistance, supporting public health initiatives.

Antimicrobial photodynamic therapy (APDT) and silver nanoparticles (AgNPs) are known as promising alternatives for the control of microorganisms. This study aims to evaluate the antifungal activity of APDT, particularly by using the association of low concentrations of phenothiazinium photosensitizers (PS) methylene blue (MB), new methylene blue N (NMBN), and new methylene blue N Zinc (NMBN-Zn) in association with biosynthesized AgNPs. The AgNPs were characterized by UV-Vis spectrophotometry, transmission electron microscopy, and the dynamic light scattering method. The minimum inhibitory concentration of compounds in APDT against Candida albicans and Fusarium keratoplasticum was obtained and the Fractional Inhibitory Concentration Index determined the antifungal effect. The toxicity of compounds and associations in APDT were evaluated in Galleria mellonella. The AgNPs presented a surface plasmon band peak at 420 nm, hydrodynamic diameter of 86.72 nm, and zeta potential of −28.6 mV. AgNPs-PS showed a wider and displaced plasmon band peak due to PS ligands on the surface and decreased zeta potential. AgNPs-NMBN and AgNPs-NMBN-Zn associations presented synergistic effect in APDT with 15 J cm⁻² against both fungi and did not show toxicity to G. mellonella. Hence, the enhancement of antifungal activity with low concentrations of compounds and absence of toxicity makes APDT with AgNPs-PS a promising therapeutic alternative for fungal infections.

The aim of this study was to compare the performance of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), phenotypic and molecular methods for the identification of Fusarium species complexes isolated from clinical cases in the State of Sao Paulo (Brazil) between the years 2001 and 2017. Sequencing of ITS region of ribosomal DNA and elongation factor 1 alpha gene (ET1α) were used as reference method in the analysis of a total of 108 Fusarium spp. clinical strains isolated from human hosts with superficial and systemic infections. Agreement between MALDI-TOF-MS and molecular data was observed for 97 out of 108 clinical isolates (89.8%), whereas five (4.6%) and six (5.5%) clinical isolates were misidentified and were not identified by MALDI-TOF MS, respectively. ITS region sequences and MALDI-TOF MS mass spectra identified and grouped correctly most of Fusarium clinical isolates at species complex level. This investigation highlights the potential of MALDI-TOF MS technique as a fast and cost-efficient alternative for clinical Fusarium identification. However, MALDI-TOF MS requires a more accurate and larger database. This work is the first comprehensive report for Fusarium population, based on phenotypic analyses, proteomic profile by MALDI-TOF and phylogenetic analyses of Fusarium species complexes isolated from clinical cases in the State of Sao Paulo, Brazil.

Disseminated fusariosis is treated with amphotericin B and voriconazole. To determine adequate therapy, the minimal inhibitory concentration (MIC) is used. However, MIC analysis is based on visual observation and requires a long period of fungal incubation. The measure of the minimal profile change concentration (MPCC) using MALDI-TOF MS is a quick spectral method that has presented good results in determining the antimicrobial resistance of yeasts. However, there is a lack of information on filamentous fungi. In the present work, 13 Fusarium spp. clinical isolates and two reference strains were used. MIC was obtained according to the M38-A2 protocol of the Clinical Laboratory Standards Institute, while MPPC was obtained following the initial steps of the M38-A2 protocol. Both Biotyper and the Rstudio environment were used to analyze mass spectra. For some fungal strains, the data obtained from the software MALDI Biotyper Compass 4.1 led to fuzzy heatmaps resulting in difficult interpretation, while heatmaps obtained using Rstudio tools generated better MPCC resolutions. Herein, 86.6% of the AMB MPCC values were highly correlated with the gold-standard AMB MIC. MALDI-TOF MS is a prominent tool used to determine MPCCs quicker, cost-effectively, and more accurately for Fusarium spp. strains. However, better statistical analyses could help measure the technique’s limit detection.

Fungal secondary metabolites display remarkable chemical diversity and biological potential, with applications in agriculture and pharmaceuticals. This study isolated and characterized bioactive metabolites from two phytopathogenic fungi, Fusarium falciforme (from papaya) and Aspergillus terreus (from pineapple), assessing their antifungal and cysteine protease‐inhibitory activities. The compounds hymeglusin, fusaridioic acid A, and butyrolactone I exhibited significant antifungal effects, with hymeglusin (IC ₅₀ 22.12 µg/mL) inhibiting A. terreus and butyrolactone I (IC ₅₀ 39.72 µg/mL) inhibiting F. falciforme . These metabolites also strongly inhibited papain, with IC ₅₀ values of 6.24 µM (butyrolactone I), 9.38 µM (hymeglusin), and 28.58 µM (fusaridioic acid A). Butyrolactone I exhibited the highest antitrypanosomal activity (IC ₅₀ 24.72 µM), highlighting its antiparasitic potential. These findings emphasize the biotechnological significance of fungal metabolites for antifungal, protease‐inhibitory, and antiparasitic applications while also shedding light on their ecological role in host–pathogen interactions.