SCREENING METHODS: Antifungal assays(2)

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A well is then bored in the centre of the agar and test substance added to the well, or the test substance is added to a paper disk and the disk placed in the centre of the agar. (The specific agar to be used, and temperature and time of incubation, will be determined by the fungi to be used.) The growth of the fungi is monitored and any inhibition of mycelia growth noted. This inhibition of growth is then expressed as a percentage of the growth of control colonies.

In the agar dilution method (also known as the poison food technique) the test substance is incorporated into the agar substrate and then a sample of actively growing fungus is placed at the centre of the plate. The radial growth of the fungus after an appropriate time, depending on the growth characteristics of the fungus, is then measured and compared with control samples. Sridhar et al. (2003) used this method to show the activity of essential oils against a range of fungi of agricultural and medical importance. Alternatively a fungal cell suspension may be inoculated onto the plate and the MIC determined by the lowest concentration of test substance that prevents visible fungal growth de Aquino Lemos et al. (2005).

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