Hi Everyone,
For my current project, I am growing bacteria and fungi together to mimic the gut microbiota (cultured from faeces). I am looking for a protocol to evaluate fungal growth from a sample of the medium. Hence, I am looking for something similar to SCFA production, lactate production, gas production, etc... basically by-products quantification specific for fungal growth.
Since fungal growth is correlated with chitin and chitosan production (cell wall components), I was thinking about an assay to evaluate its quantification but I have not found anything robust so far. Most of the assay use an hydrolysis step to release GlcN. However, GlcN is also present in bacteria hence, it would not be specific to Fungae.
I have also identified to use of Calcofluor White staining which also stains the chitin and which can be used on flow cytometry. Anyone has knowledge on the technic? Can it be used for fungal quantification?
Do you have anything to suggest?
Kind regards,
Hi Florian,
Thanks for your answer. I am already thinking of using qPCR, and illumina sequencing as well. However, they use DNA extraction, which - for now - have controversial effect on the obtained results (the recovery is not yet well determined and strong biaises have been recently reviewed depending on the method applied which favours few species than others). Also, those method are having a stringent threshold, which is not great for the mycobiota supposed to be in low amount in the gut (that's why I am having concerns for the flow cytometry method as well).
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