i have methonolic extract of fungus and i want to do antimicrobial activity. what is the best procedure to follow disc diffusion or agar well method or in what quantity extract is use.
Dear Vipin,
May I suggest you to refer to antimicrobial activity monograph from IP wherein you will get all the details of both antibacterial and antifungal preparations with every possible details related to the topic under consideration.
Regards and best wishes,
Dr. A.D. Padhye.
For the methanolic extract you should go for well diffusion method and keep one control well filled with specified quantity of pure methanol solvent without fail.
Concentration of extract may vary.
You have to standardize it.
For the 1st assessment set use most lower, most higher and middle concentration of the extract (in accordance with the literature of similar studies).
All the best
I prefer to start with agar well diffusion method by testing several concentrations on more than one type of media (nutrient, PDA, PCA..... etc.).
For anti-fungal activity its possible to use micro dilution technique.
Use serial dilution to calculate minimum inhibitory concentration (MIC).
Good Luck.
Hi,
Please read my papers about ionic liquids and oxidative stress. You will see how you can prepare antimicrobial tests not based on disc diffusion which can fail. If you have any further questions please do not hesitate to contact me.
Cheers,
Tomasz
Dear Vipin,
Based on my earlier experience with anti-microbial activity test of a microbial secretion I suggest you should go for the disc-diffusion test first because of its simplicity. You should prepare a lawn of the target microorganism by spread plate method and place the discs impregnated with your fungal extract with varying concentration (to detect the MIC, if any). Do not forget to place a control disc for comparison. You will get the complete instructions if you search for "Kirby-Bauers Disc diffusion method". After getting desirable results you can always go for well diffusion for a double check.
Best wishes,
Shiladitya
Vipin,
I would also start with the Kirby-Bauer method and do a disc diffusion on an agar plate. If you used the large plates you can get a number of results at one time: http://www.microbeworld.org/component/jlibrary/?view=article&id=12255. You can also do the E test: http://www.microbeworld.org/component/jlibrary/?view=article&id=13789
Cheers,
Tasha
Etest is a concentration gradient of the antibiotic on a single strip to determine MIC. A manufactured strip but you can do the same thing using sterile disc at varying concentrations of the methonolic extract.
Thanks Vipin and Tasha,
I was also wondering about the term: Etest. I am familiar with the procedural part but it would be nice if you could expand the acronyme Etest.
Regards,
The Epsilometer test......I had to look it it since I have always known it as the "Etest" and forgotten what it stood for.
Cheers,
Tasha
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