| Congresso Brasileiro de Microbiologia 2023 | Resumo: 1235-1 | ||||
Resumo:In recent years, a notable increase in fungal infections caused by Candida species has been observed. This growth can be attributed to various factors, including therapeutic limitations, an increase in immunocompromised patients, indiscriminate use of antimicrobials, and an aging population. Consequently, the search for new antifungal agents has become of utmost importance. Punica granatum, commonly known as pomegranate, is a small fruit-bearing tree belonging to the Lythraceae family. Both in vitro and in vivo studies have been conducted to explore the antifungal potential of compounds present in pomegranate against various pathogenic fungi, particularly Candida species. In this context, the objective of this project was to investigate the antifungal activity of pomegranate extracts and evaluate the mechanism of action through assays involving the use of sorbitol, ergosterol, and membrane permeability evaluation in relation to the mentioned Candida species. Standard strains of Candida albicans (ATCC24433), Candida krusei (ATCC20298), Candida parapsilosis (ATCC22019), and Candida glabrata (ATCC2001) were used. The determination of the minimum inhibitory concentration (MIC) was performed using the microplate dilution methodology, following the Clinical and Laboratory Standards Institute (CLSI) M27-A3 2008 guidelines. The sorbitol assay was conducted in the same manner as MIC determination, with the addition of 0.8 M sorbitol. The ergosterol assay involved the use of concentrations of 50, 100 and 200 µg/mL of ergosterol. Lastly, membrane permeability evaluation was performed through the crystal violet assay, where crystal uptake increases if there is a change in cell permeability to concentrations of 1x MIC and 2x MIC of pomegranate extract. The results revealed that Punica granatum extract presented an MIC of 10 mg/mL against the tested fungi. However, the assays conducted to investigate the mechanism of action did not demonstrate any possible explanation for the activity of the extract. The MIC value was not altered with the use of sorbitol against any strain, suggesting that the extract does not directly act on the fungal cell wall. Additionally, the results did not indicate any interaction of the extract with fungal cell membrane ergosterol. In the membrane permeability test, a low absorbance uptake was observed in all Candida strains, including Candida albicans, where the absorbance was 23.25 and 27.78 at concentrations of 1x MIC and 2x MIC, respectively. This led to the conclusion that the extract did not interfere with the integrity of the fungal cell membrane. A possible explanation for these results may be related to pomegranate harvest factors, which can play a significant role in the chemical composition and bioactive properties of the plant. Therefore, further studies are suggested to better understand the mechanism of action of this extract and its potential efficacy as an antifungal agent. These studies may provide valuable information for the development of new therapeutic approaches in combating fungal infections. Palavras-chave: Punica granatum, antifungal activity, Candida, mechanism of action Agência de fomento:Fapes | |||||