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hermit
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Registered: 08/14/06
Posts: 35
Loc: EU, zone 4
Last seen: 16 years, 1 month
Cleaning contaminated spores?
    #6330123 - 12/03/06 12:47 PM (18 years, 1 month ago)

I have a fresh outdoors azurescens print (from FSRE, thanks!) which unfortunately appears hopelessly contaminated with mold spores.
Here's my first attempt on potato-carrot agar:



Is there any way to wash or treat mushroom spores to reduce mold spore load? Anything that kills more mold spores than mushroom spores? Dilute H2O2? (How dilute?) Acetic or citric acid? (How strong?) Heat? (What temperature, how long?) Any advice?

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Invisibleroach6
ethnobotanist
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Re: Cleaning contaminated spores? [Re: rip]
    #6330147 - 12/03/06 01:03 PM (18 years, 1 month ago)

I dont see any mushroom mycelium on that plate. Toss it before ou have a housefull of bad spores.


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hermit
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Re: Cleaning contaminated spores? [Re: roach6]
    #6330391 - 12/03/06 02:49 PM (18 years, 1 month ago)

I know there is no mycelia. That's why I'm asking about cleaning the spores _before_ putting them on agar because it is obviously pretty hopeless after germination for these. Sorry if I did not make myself clear.

(And yes, I did dispose these petris unopened immediately after taking the picture.)

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hermit
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Re: Cleaning contaminated spores? [Re: rip]
    #6372952 - 12/15/06 07:44 AM (18 years, 1 month ago)

This is my second attempt. Procedure: mix the spores with 0.3% diluted bleach in a test tube, let settle one hour in room temperature, remove most of the bleach solution with a syringe, mix with 1ml of sterile water and spread on potato carrot sawdust agar. I do not know what germinating azu mycelium should look like, but this looks much better than my first attempt. (Anyone?)




I tried 3%, 1%, 0.3% and 0.1% bleach. I used the cheapest brand in the shop, 5% or less NaOCl, so my solutions contained roughly 20, 6, 2, and 0.6 mMol/l or less of NaOCl (if I calculate it right). After one week incubation, the 3% and 1% treated spores appear all dead and both the 0.3% and 0.1% germinate with no apparant difference in germination rate. I used too much spores to make a colony count, about 1/8th of a large print for 4 portions. If anyone is interested of replicating this, I'd suggest using much less. (These were fresh spores.)

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InvisibleCureCat
Strangest
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Registered: 04/19/06
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Re: Cleaning contaminated spores? [Re: rip]
    #6373928 - 12/15/06 12:23 PM (18 years, 1 month ago)

Interesting... Not sure how to help you, I'm not agar savvy, but i'll be interested to see how this goes.  :smile:


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hermit
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Registered: 08/14/06
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Loc: EU, zone 4
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Re: Cleaning contaminated spores? [Re: CureCat]
    #6375854 - 12/16/06 01:01 AM (18 years, 1 month ago)

Well, I've transferred samples of the assumed azu mycelia to agar, sterilized woodchips and soaked corrucated cardboard. Time will tell. (I'll post if it turns out being something else...) No contaminants spotted yet.

It is bit early to say, but I think washing spores with dilute bleach might make possible to use wild collected outdoors spores even for direct spore syringe inoculation of substrate without going agar.

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OfflineFamiliar Gnome
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Re: Cleaning contaminated spores? [Re: rip]
    #10609357 - 07/02/09 11:49 AM (15 years, 6 months ago)

This is an old thread... how did it go finally?

  I used this TEK to clean some prints I got from a friend, at 0.3% it worked like a charm!

Thanks.


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