SCREENING METHODS: Poison food technique

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Generally antifungal activity is determined by poisoned food technique (Nene and Thapliyal, 2000, Kiran et al., 2010). Five-day old fungal culture is punched aseptically with a sterile cork borer of generally 6mm diameter. The fungal discs are then put on the gelled agar plate. The agar plates have been prepared by impregnating desired concentration of plant extract at a temperature of 45-50 °C. The plates are then incubated at temperature 26 ± 2°C for fungi. Colony diameter is recorded by measuring the two opposite circumference of the colony growth.

Percentage inhibition of mycellial growth is evaluated by comparing the colony diameter of poisoned plate (with plant extract) and nonpoisoned plate (with distilled water) and calculated using the formula given below (Verma and Kharwar, 2008);

Screening Methods in the Study-1

Spore germination assay is a slide technique used for testing of antifungal activity of plant extracts (Nair et al., 1991). Plant extract of desired concentration and volume are added to the surface of dried slides as a film or in a cavity of a cavity slide. Fixed volume and standard concentration of spore suspension of test fungi are spread over the film whereas in controlled treatment, distilled water is added in place of spore suspension. Slides are then placed on a glass rod in Petri dish under moistened conditions and incubated for 26 ± 2°C. After incubation, slides are fixed in lacto phenol cotton blue and observed microscopically for spore germination. Percentage spore germination is calculated according to the following formula.

% Spore germination = Number of germinated spores / Total number of spores X 100

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