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OfflineExplosiveMango
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Recovering from Contam with Agar
    #6132251 - 10/04/06 12:37 PM (17 years, 5 months ago)

Right now I have a friend who is using agar to recover mycelium from a contaminated spore bank.

Procedure:
-Inoculate straight malt agar with contaminated supply

-Wait for mycelium growth and transfer to peroxidated malt agar

-Repeat if necessary






What I'm wondering is this:
Is it smarter to transfer the mycelium from agar to agar as soon as it forms and travels away from the contam? Or should some time be given to the mycelium to gain strength before transferring it into a peroxidated culture?

Thanks.


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OfflineExplosiveMango
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Re: Recovering from Contam with Agar [Re: ExplosiveMango]
    #6132569 - 10/04/06 02:02 PM (17 years, 5 months ago)

The specific contaminant appears to be green mold.


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OfflineBabo911
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Re: Recovering from Contam with Agar [Re: ExplosiveMango]
    #6132585 - 10/04/06 02:07 PM (17 years, 5 months ago)

as soon as you see new growth put it on another plate of agar and repeat as needed.

The H2O2 agar after you cut a piece off the multi-spore is a great idea IMOP


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OfflineRogerRabbitM
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Re: Recovering from Contam with Agar [Re: Babo911]
    #6132875 - 10/04/06 03:31 PM (17 years, 5 months ago)

Antibiotic agar will be much more effective than peroxide. Either one is for control of bacteria however. Peroxide is not the best choice for helping your mycelium outrun another fungi. Mushroom mycelium can usually recover from peroxide, but when isolating from contamination on agar, you want your mycelium to run like fire, so skip the peroxide.

With a flame sterilized scalpel, transfer each new growth the minute you see it and can differentiate it from other growths. Transfer each and every growth you see to its own petri dish. This means you'll have some dishes that turn out to be 100% mold, but also dishes that will be pure mushroom mycelium, provided you can get a clean transfer.

If you wait too long, the mold will sporulate, then when you transfer the good stuff, it will have mold spores attached, ruining the culture later on.

It should go without saying to use a glovebox. If a flowhood is used, the airflow will blow the contaminant spores all over the place. This is just a note to those of you who have just purchased or made a flowhood. Never open a contaminated petri dish or jar in front of it.
RR


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