In-Vitro Antifungal Assessment Drug Research Study
Project Description
in vitro anti fungal assessment (MIC/MFC) on various fungal strain using a clotimazole as standard anti-fungal drug. :
Trichophyton erinacei
Trichophyton interdigitale
Trichophyton rubrum
Trichophyton schoenleinii
Trichophyton tonsurans
Trichophyton eboreum
Trichophyton concentrium
Trichophyton mentagrophytes
Trichophyton tonsurans
Malassezia furfur
+ using a clotrimazole as standard anti fungal drug
+ Determination of the antifungal activity.
In vitro anti-Trichophyton assay:
MIC determination
Plant extracts were serially diluted using Sabouraud dextrose broth (SDB) by "serial dilution technique". Twenty microliter of inoculum was added. SDB containing only 20 ml of inoculum served as positive control, SDB alone served as negative control. The tubes were incubated at (35 ± 2°C) for 20 days. The MIC was the lowest concentration of the extract that did not permit any visible growth. [1]
For Malassezia furfur
To assess the antifungal activity of the plant, the agar-well difusion method was used. Modified Dixon's agar was utilized as a culture medium. Fourteen milliliters of Dixon medium were inoculated with 1 ml of a suspension of M. furfur (105 CFU/ml in phosphatebuffered saline (PBS) 1 X) in Petri dishes. Inoculums suspensions were previously adjusted by spectrophotometry to an absorbance of 0.1 at 530 nm.
The antifungal activity was evaluated by measuring the inhibition-zone diameter observed after 48 h of incubation using a clotrimazole as standard drug. [2]
For MFC determination [1], 21 days old incubated (MIC) suspension were subcultured in SDA plates using an inoculum size of 1 ml, and were incubated at room temperature for 21 days. The MFC was recorded as the lowest concentration that prevented the growth of any fungal colony on the solid media.
The minimum inhibitory concentration (MIC) was defined as the lowest concentration of the extract to inhibit the growth of microorganisms.
The minimum fungal concentration (MFC) was defined as the concentration of extract that did not show any microbial growth.
Refrances
1. Vaijayanthimala, J., Prasad, N.R., Anandi, C. and Pugalendi, K.V., 2004. Anti-dermatophytic activity of some Indian medicinal plants. Journal of natural remedies, 4(1), pp.26-31.
2. Filip, R., Davicino, R. and Anesini, C., 2010. Antifungal activity of the aqueous extract of Ilex paraguariensis against Malassezia furfur. Phytotherapy Research: An International Journal Devoted to Pharmacological and Toxicological Evaluation of Natural Product Derivatives, 24(5), pp.715-719.
Project Information
Number:20-01691
Industry:Pharmaceuticals
