Withania somnifera (L.) Dunal whole-plant extracts exhibited anti-sporotrichotic effects by destabilizing peripheral integrity of Sporothrix globosa yeast cells

Abstract

Chronic topical cases of Sporotrichosis, a chronic fungal infection caused by the ubiquitously present cryptic members of the Sporothrix species complex, are treated with oral administrations of itraconazole. However, severe pulmonary or disseminated cases require repeated intra-venous doses of amphotericin B or even surgical debridement of the infected tissue. The unavoidable adverse side-effects of the current treatments, besides the growing drug resistance among Sporothrix genus, demands exploration of alternative therapeutic options. Medicinal herbs, due to their multi-targeting capacity, are gaining popularity amidst the rising antimicrobial recalcitrance. Withania somnifera is a well-known medicinal herb with reported antifungal activities against several pathogenic fungal genera. In this study, the antifungal effect of the whole plant extract of W. somnifera (WSWE) has been explored for the first time, against an itraconazole resistant strain of S. globosa. WSWE treatment inhibited S. globosa yeast form growth in a dose-dependent manner, with IC50 of 1.40 mg/ml. Minimum fungicidal concentration (MFC) was found to be 50 mg/ml. Sorbitol protection and ergosterol binding assays, revealed that anti-sporotrichotic effects of WSWE correlated well with the destabilization of the fungal cell wall and cell membrane. This observation was validated through dose-dependent decrease in overall ergosterol contents in WSWE-treated S. globosa cells. Compositional analysis of WSWE through high performance liquid chromatography (HPLC) exhibited the presence of several anti-microbial phytochemicals like withanone, withaferin A, withanolides A and B, and withanoside IV and V. Withanone and withaferin A, purified from WSWE, were 10–20 folds more potent against S. globosa than WSWE, thus, suggesting to be the major phytocompounds responsible for the observed anti-sporotrichotic activity. In conclusion, this study has demonstrated the anti-sporotrichotic property of the whole plant extract of W. somnifera against S. globosa that could be further explored for the development of a natural antifungal agent against chronic Sporotrichosis. Author summary Sporotrichosis, commonly known as ‘rose gardener’s disease’ is a rare but chronic fungal infection caused by several pathogenic members of Sporothrix. Although rare, Sporotrichosis can spread to the lungs or whole body, and thus, could be fatal. However, due to low frequency of its occurrence, scientific research on Sporotrichosis has been rather limited. To exacerbate the situation, many Sporothrix members have developed resistance against the common anti-fungal agents used to treat fungal infections. Many medicinal herbs are being explored for their effectiveness against drug-resistant microorganisms. In this connection, the well-known medicinal plant, Withania somnifera, commonly known as Winter cherry in English and Ashwagandha in Hindi, has been explored for its anti-fungal potentials against several pathogenic fungi. However, to the best of our knowledge, anti-fungal effect of W. somnifera against Sporothrix has not been assessed yet. Therefore, this study was conducted to investigate the anti-fungal potentials of Withania somnifera whole plant extract (WSWE) against Sporothrix globosa, known to cause Sporotrichosis in Asia. This study demonstrated significant anti-sporotrichotic effect of WSWE, which could be further explored for developing an alternative natural treatment for Sporotrichosis.