I am acquiring some blank loads for the Kirby Bauer test so I can make my own antibacterial extracts for the antibiotic disks. Because I will be dealing with a broad range of fungi I am searching for a general protocol to start. Does anyone have a preferred method?
Here is a report of a rapid, effective and reproducible assay to detect fungal antimicrobial activity ....of fungal properties against single cell microorganisms This method allows an easy way of allowing an easy way of performing a fast antimicrobial screening of actively growing fungi directly against yeast. Because it makes use of an actively growing mycelium, this bio-assay also provides a way for studying the production dynamics of antimicrobial compounds by filamentous fungi. The proposed assay is less time consuming and introduces the innovation of allowing the direct detection of fungal antimicrobial properties against single cell microorganisms without the prior isolation of the active substance(s). This is particularly useful when performing large screenings for fungal antimicrobial activity. For more details consult https://www.sciecncedirect.com or Microbigical Research Vol. 168, Issue 1, 15 January 2013,pages 1 - 64
I have used methods in the past such as the one described in this paper: Article Discovery and Biosynthesis of Gladiolin: A Burkholderia glad...
.For looking at a broad range of compounds it may be easier to use a solvent extraction method. I used DCM due to its medium polarity and therefore would extract many compounds.
When purifying extracts you can use column chromatography for a quicker method.
Looking at the antimicrobial activity you can use an overlay assay as this may be quicker?
Hope this helps!
Thank you all, Andrew Angus John Robertson I really enjoyed reading the Discovery and Biosynthesis of Gladiolin. This is very impressive! The MIC values, REMA assay, SMRT sequencing and purification methods used in this publication will push me in the right direction. The American Chemical Society, CLSI, and NIH have been a guiding light in this research project. I greatly appreciate you sharing and would love to hear more about your personal research if you ever want to chat about it! Thanks friend...Have you ever read CLSI M51-A ? I am trying to purchase a copy before I start the experiment, here is a link to the sample booklet: https://clsi.org/media/1647/m51a_sample.pdf
I think a copy of this would allow my experimentation to be as accurate as possible not to mention it is the standard way of doing things, when i get my hands on a copy i would be happy to share if you are interested!
This may help you Article Antimicrobial activity of crude extracts prepared from fungal mycelia
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