We isolated some endophytic fungus and bacteria from plant source. Now we want to check their antibacterial and antifungal activity . I have found Agar overlay technique in some paper. How effective is this agar overlay technique ? What is the drawback of this technique and if there is any better technique to initially checking of antibacterial activity, please suggest.
The best method is disc diffusion method for initial screening. you can follow either EUCAST or American standards. But there is a problem for non polar bio-active metabolites since those can not be diffuse through the polar agar medium. Therefor more sensitive method is MIC, but it is time consuming. My opinion is first perform a disc diffusion assay and then select the active extracts and if you want to see the more potent extract then do the MIC only for the active extracts. The well diffusion method is encountered with lots of errors there for not much suitable for initial screening. For anti-fungal assay most reliable method is poison food method but for that you want little bit higher amount of the sample. But for the initial screening disc diffusion method is also accepted.
Dear Badrul,
To check antibacterial activity Agar well diffusion and Broth dilution methods are well recommended.......
For any method Mueller-Hinton agar or broth medium is well accepted due to diffusion of your test organism extract.....
Broth dilution method is normally done with microtitre plate......here less amount of your extract is consumed....we can test on many pathoges......at a time....MIC and MBC can be estimated very well with broth dilution method......
Compare to agar well and Disk diffusion I recommend broth dilution method.........
All the best
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