Six sterile porcelain cylinders with outside diameter of 8 mm and length 10 mm were placed on the base layer about 608 apart before pouring the seed layer. Then, the seed layer was prepared by adding the determined quantity of the test organism to the medium at a temperature of 40 and 50 8C and immediately poured the inoculated medium on the top of base layer. After solidifying the seed layer, the cylinders were carefully removed using a sterile forceps so that the uniform cavities were formed in the seed layer. Fifty microliters each of reference standard (10 mM amphotericin) and test compound were added in each well. Three alternate wells on each plate were filled with a reference standard and the remaining three were filled with test solution. The reference standard was used for the verification of strain sensitivity/resistance pattern. Inhibition area was measured after overnight incubation at 30 C. Screening for antifungal activity