Synergistic activity between conventional antifungals and chalcone-derived compound against dermatophyte fungi and Candida spp.

The scarce antifungal arsenal, changes in the susceptibility profile of fungal agents, and lack of adherence to treatment have contributed to the increase of cases of dermatomycoses. In this context, new antimicrobial substances have gained importance. Chalcones are precursors of the flavonoid family that have multiple biological activities, have high tolerability by humans, and easy synthesis. In this study, we evaluated the in vitro antifungal activity, alone and in combination with conventional antifungal drugs, of the VS02-4'ethyl chalcone-derived compound against dermatophytes and Candida spp. Susceptibility testing was carried out by broth microdilution. Experiments for determination of the target of the compound on the fungal cell, time-kill kinetics, and toxicity tests in Galleria mellonella model were also performed. Combinatory effects were evaluated by the checkerboard method. Results showed high activity of the compound VS02-4'ethyl against dermatophytes (MIC of 7.81-31.25 µg/ml). The compound targeted the cell membrane, and the time-kill test showed the compound continues to exert gradual activity after 5 days on dermatophytes, but no significant activity on Candida. Low toxicity was observed at 250 mg/kg. Excellent results were observed in the combinatory test, where VS02-4'ethyl showed synergistic interactions with itraconazole, fluconazole, terbinafine, and griseofulvin, against all isolates tested. Although further investigation is needed, these results revealed the great potential of chalcone-derived compounds against fungal infections for which treatments are long and laborious.

Antifungal activity of 3,3'-dimethoxycurcumin (DMC) against dermatophytes and Candida species.

Dermatomycosis is an infection with global impacts caused especially by dermatophytes and Candida species. Current antifungal therapies involve drugs that face fungal resistance barriers. This clinical context emphasizes the need to discover new antifungal agents. Herein, the antifungal potential of 10 curcumin analogs was evaluated against four Candida and four dermatophyte species. The most active compound, 3,3'-dimethoxycurcumin, exhibited minimum inhibitory concentration values ranging from 1.9‒62.5 to 15.6‒62.5 µg ml-1 against dermatophytes and Candida species, respectively. According to the checkerboard method, the association between DMC and terbinafine demonstrated a synergistic effect against Trichophyton mentagrophytes and Epidermophyton floccosum. Ergosterol binding test indicated DMC forms a complex with ergosterol of Candida albicans, C. krusei, and C. tropicalis. However, results from the sorbitol protection assay indicated that DMC had no effect on the cell walls of Candida species. The in vivo toxicity, using Galleria mellonella larvae, indicated no toxic effect of DMC. Altogether, curcumin analog DMC was a promising antifungal agent with a promising ability to act against Candida and dermatophyte species.

Combinatorial effect of fluconazole, itraconazole, and terbinafine with different culture extracts of Candida parapsilosis and Trichophyton spp. against Trichophyton rubrum.

Onychomycosis is a nail infection caused by dermatophytes, non-dermatophyte fungi, and yeasts, especially Candida species. The present study evaluated the combinatorial effect of different cultured extracts of Candida parapsilosis and Trichophyton mentagrophytes and Trichophyton rubrum with fluconazole, itraconazole, and terbinafine against clinical isolates of Trichophyton rubrum. In addition, investigation of the action of the extracts on the wall or membrane was performed. Pure and mixed cultures of Candida parapsilosis and dermatophytes were filtered through a 0.2-µm membrane and submitted to liquid-liquid extraction using ethyl acetate. After a checkerboard, trial with drugs was performed to evaluate the synergistic interaction with the extract. The results obtained for the minimum inhibitory concentration (MIC) of extracts against the T. rubrum strain in isolation were 500-8000 µg/mL. The MIC range for fluconazole, itraconazole, and terbinafine were 2-32 µg/mL, 0.25-0.5 µg/mL, 0.03-64 µg/mL, respectively. However, when the extract was combined with drugs, the MIC values decreased: extracts 1.9-1000 µg/mL, fluconazole 0.25-4, itraconazole 0.03-0.06 µg/mL, and terbinafine 0.001-0.02 µg/mL. The MIC values of the extracts in the Roswell Park Memorial Institute 1640 medium (RPMI) supplemented with sorbitol did not change, suggesting any action on the cell wall. However, in the presence of RPMI supplemented with ergosterol, MIC values of the extracts increased by up to 2×, indicating action on the fungal cell membrane. A synergistic action was observed between products and drugs, detecting a decrease in MIC values. There is potential and a new therapeutic perspective for fungal control.

Sporothrix pathogenic clade: Molecular analysis of animal and human clinical isolates.

Sporotrichosis is a subcutaneous mycosis that affects animals and humans. Varying in severity, occurrences range from local lesions to systemic involvement. It is caused by thermodimorphic and saprobic fungi from the Sporothrix pathogenic clade. This study aimed to identify the species and the sexual idiomorph distribution patterns responsible for diagnosed cases of sporotrichosis in São José do Rio Preto, Brazil. We included 188 isolates of Sporothrix sp. from feline lesions and 27 of human origin, which underwent molecular identification and genotyping for mating-type MAT1-1 and MAT1-2. The results showed that Sporothrix brasiliensis is the prevalent species in feline sporotrichosis outbreaks with the overwhelming presence of a single mating-type, MAT1-2 (P <.0001), suggesting a prevalently clonal form of spread. Morphological analyses did not discriminate among cryptic species in the genus Sporothrix, and molecular identification was essential for the correct identification of the species responsible for the observed cases of sporotrichosis. Distribution analyses of MAT1-2 isolates support the hypothesis of unidirectional migration from the current epidemics in São Paulo and Rio de Janeiro to the municipality of São José do Rio Preto.

This study aimed to identify the species and the sexual idiomorph distribution patterns responsible for diagnosed cases of sporotrichosis in São José do Rio Preto, Brazil. We included 188 isolates of Sporothrix sp. from feline lesions and 27 of human origin.

Clinical and epidemiological aspects of feline sporotrichosis caused by Sporothrix brasiliensis and in vitro antifungal susceptibility.

Sporotrichosis is a subcutaneous mycosis resulting from the traumatic implantation of pathogenic Sporothrix species. In Brazil, zoonotic transmission plays an important role in the epidemiology of the disease, involving especially cats. The objective of this study was to isolate Sporothrix spp. from cats with signs of sporotrichosis, determining the causative species, clinical and epidemiological aspects, and the in vitro susceptibility profile of the isolates against antifungal drugs. From September 2017 to February 2019, 245 samples of lesions were collected from symptomatic cats in São José do Rio Preto, Brazil. Identification of the isolates was performed by morphophysiological parameters and species-specific polymerase chain reaction. The susceptibility profile of the isolates was determined for five drugs (amphotericin B, itraconazole, ketoconazole, potassium iodide and terbinafine), using the broth microdilution method. Clinical and epidemiological aspects were analyzed based on data contained on investigation forms filled by the veterinarians at moment of collection. Sporothrix spp. were isolated in 189 (77.2%) of the samples. Phenotypic and molecular analyses revealed S. brasiliensis as the only causative agent. In vitro susceptibility testing showed lower MIC values for terbinafine (MIC = 0.03-2 µg/ml), ketoconazole (MIC = 0.03-2 µg/ml), and itraconazole (MIC = 0.03-4 µg/ml). Most of the animals were male (73.5%), adults (96.3%), stray (53.5%), and uncastrated (69.8%). Our results show the expansion of the S. brasiliensis epidemic to an area nearly 840 km apart from the epicenter of the long-lasting outbreak of cat-transmitted sporotrichosis in Rio de Janeiro.