I want to test the production of mycotoxins by some mould isolates. Will they (if capable) produce the toxins if grown as a pure culture in broth media suitable for their growth?
In addition to answer from Thippeswamy, you need to search literature for the best suitable medium for cultivating the mould to get the required mycotoxin(s) and the best analytical platform (extraction, clean-up and estimation) for analysis of toxin(s). If you are working with a yet-to-be identified species of mould and you need to assy for mycotoxin production, then use a fairly flexible medium (YES for Aspergillus and Penicillium species, maize grains for Fusarium species). All these are dependent on the fungal species and your target mycotoxin.
BY DIRECT PLATING AND DILUTION PLATING METHODS.
OTHER METHODS USED ARE EIA . LEVEL OF TOXINS IN HUMANS CAN BE DETECTED BY EXAMINATION OF URINE, BLOOD, BODY FLUIDS.
WE HAVE ALSO FACED WITH SAME SITUATION MANY TIMES. IT IS REALLY DIFFICULT TO EXPLAIN THE ROLE OF THESE UNUSUAL FUNGI. WE DO FIND SEVERAL DEMATACIOUS FUNGAL ISOLATES FROM SKIN AND NAILS.
First you have to extract the toxin from broth culture after 7-10 days of incubation using suitable solvents, for instance you can use chloroform for extraction of aflatoxin, then you can spot extracted toxin on TLC plate along with standard toxin and observe under UV light or you can observe by spraying with suitable reagent or you can also use HPLC for detection.
In addition to answer from Thippeswamy, you need to search literature for the best suitable medium for cultivating the mould to get the required mycotoxin(s) and the best analytical platform (extraction, clean-up and estimation) for analysis of toxin(s). If you are working with a yet-to-be identified species of mould and you need to assy for mycotoxin production, then use a fairly flexible medium (YES for Aspergillus and Penicillium species, maize grains for Fusarium species). All these are dependent on the fungal species and your target mycotoxin.
Check the work of Jens Frisvad from Denmark. You can simply place plugs of agar on a TLC plate and develop the plate in solvent, etc. You can extract the plugs in solvent too and undertake HPLC. I have published on this too (see Paterson R R M in various publications). A lot of this work started in the 1980s but it is still valid and had been updated.
Spot the extracted toxin on TLC plate and observe under UV light or you can use HPLC for detection. However all these are dependent on the fungal species and your particular required mycotoxin.
The only other thing I would mention is that ther are data bases of tlc and HPLC characteristics of the mycotoxins in various publications, but particularly in J Chromatography.
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