ABSTRACT
Plants have provided a source for novel drug compounds, as plant derived medicines have made large contributions to human health and well-being. The aerial parts of Mitracarpus villosus were powdered, screened for phyto-compounds and the phytocompounds detected were isolated. This was followed by successive extraction of the plant (hexane, ethyl acetate, ethanol and water) and fractionation of the ethyl acetate extract. Antifungal activities of the crude extracts, fractions and extracted secondary metabolites against clinical isolates of Candida albicans, Candida krusei, Trichophyton verrucosum, Trichophyton mentagrophytes, Aspergillus fumigatus and Aspergillus niger were investigated using agar diffusion, broth dilution and micro broth dilution methods. The antibiotic susceptibility profiles of the fungal isolates to standard antifungals such as fluconazole and ketoconazole were determined using agar diffusion and broth dilution methods. The effects of temperature and different storage conditions on the antifungal activity of the crude plant extracts, as well as the rate of death / survival of the fungal isolates on exposure to crude extract and standard drugs were also investigated. Phytochemical screening powdered plant revealed the presence of tannins, saponins, flavonoids, terpenes, phenols and resins. The ethyl acetate extract of the plant aerial parts produced the highest antifungal activity when compared with the other solvent extracts (hexane, ethanol and water) with inhibition zone diameter ranging from 18.67 – 21.67 mm at an exposure concentration of 12.5 mg/mL. The crude tannin, saponin and phenol extracts produced zones of inhibition ranging from 19.67- 24.0 mm, 19.0 –24.67 mm and 18.33- 22.0 mm respectively. The minimum inhibitory concentration and minimum fungicidal concentration values of the ethyl acetate extract against all the fungal isolates tested were 0.50 – 2.00 mg/ml and 2.00 – 8.00 mg/ml respectively. The minimum inhibitory concentrations of the crude tannins, crude saponin and phenol extracts were 0.50 – 2.00 mg/mL, 0.50 – 4.00 mg/ml and 7 0.50 – 8.00 mg/ml respectively, while their minimum fungicidal concentrations were 1.00 – 8.00 mg/ml, 1.00 – 8.00 mg/ml and 1.00 – 16.00 mg/ml respectively. The ethyl acetate fractions (M1- M6) had stronger antifungal effects on the test fungi than the crude extracts and isolated phyto-compounds. M2 and M3 had minimum inhibitory concentration values of 250.00 – 2000.00 µg/ml, M1 and M5 had minimum inhibitory concentration values ranging from 250.00 – 4000.00 µg/ml while that of M5 and M6 was 500.00 – 4000.00 µg/ml. The minimum fungicidal concentration values for M2 and M3 was 500.00 – 4000.00 µg/ml while the minimum fungicidal concentrations of fractions M1, M4, M5 and M6 ranged from 500.00 – 16000.00 µg/ml against the fungal isolates tested. Crude extracts maintained their antifungal activity within the temperature range tested and over a storage period of nine months. The pattern of kill of the fungal isolates by the ethyl acetate extract and fraction M2 of were comparable with that of the standard drugs. In conclusion, the crude extracts of Mitracarpus villosus aerial parts possess strong antifungal activities and stable over nine months storage over a temperature range of 25 – 45 oC. Further investigations should be carried out to isolate pure compounds and determine the mechanisms of action of the plant.
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