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OfflineJohn in WI
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Questions about vermiculite and contamination resistance
    #26964097 - 10/01/20 03:09 PM (3 years, 3 months ago)

I've been tinkering around with mixing spores.  I started off using prints from several strains, and inoculating agar with tiny amounts from each.  Then growing out the mycellium....

That worked, after a few transfers to make sure it was clean.  Anyway, a friend had a good idea, that perhaps I could make a spore syringe, and inoculate PF jars with it.  I tried that, and got 4 out of 8 to fruit (the other 4 falling victim to mold and being tossed).

I'm wondering what it is about BRF/vermiculite that makes it more resistant to contamination?  Also, what kind of "contamination" are we talking about?  I couldn't see bacteria, but my failed cakes had wicked mold.

The reason I'm asking is, I was wondering about using 1/2 pint jars with some vermiculite to germinate some more mixed spores from solution.  Kind a 2-dimensional PF.  Is their a carbohydrate source that would be less favorable to mold/bacteria?  Maybe instead of BRF, I could use like a 2% malt or karo solution instead?  Or a potato infusion?

If it's not possible, I can always go through the PF cake route again. I just hate throwing out that much verm, after all the work of sterilizing and everything.


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InvisiblebodhisattaMDiscordReddit
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Re: Questions about vermiculite and contamination resistance [Re: John in WI]
    #26964127 - 10/01/20 03:30 PM (3 years, 3 months ago)

Cakes don't get shook and are spongy in texture. They tend to mitigate some bacterial contaminants but not mold. I doubt it has anything to do with the ingredients but rather the construction and texture.


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OfflineJohn in WI
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Re: Questions about vermiculite and contamination resistance [Re: bodhisatta]
    #26964376 - 10/01/20 05:41 PM (3 years, 3 months ago)

ah, ok.  I was unclear, after reading for several hours.  Several people suggesting going spores to grain, some saying to germinate on cardboard...

I'm trying to think up some straight forward ways to grow small crops from very mixed spores.  I realize (and under normal circumstances I always) going to agar, transfering... is the correct way.   

But my quick and dirty PF jar run was encouraging.  I only got 50% success, but considering the original spores were old and not really properly stored, I was surprised it worked at all! 

I may just go that route.  Inoculate some PF jars, probably lose half of them, fruit out the handful that live...  I wouldn't devote a lot of resources to the technique, but it does work pretty well.


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Offlinesmalltalk_canceled
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Re: Questions about vermiculite and contamination resistance [Re: John in WI]
    #26964543 - 10/01/20 06:51 PM (3 years, 3 months ago)

you mixing strains to create a syringe to inoculate pf tek? sounds very hard and counter intuitive to me.

learn agar instead, much better


Edited by smalltalk_canceled (10/01/20 06:52 PM)


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OfflineJohn in WI
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Re: Questions about vermiculite and contamination resistance [Re: smalltalk_canceled]
    #26964673 - 10/01/20 07:58 PM (3 years, 3 months ago)

I do agar all the time for my main project.  The entire point of this side project is to produce the maximum diversity in fruits.  For the other project, I'm probably 10 transfers in, then going agar to master, then g2g.

But, if you want to go directly from MS syringe to fruit, PF cakes are not "hard".  It's a bit of a crapshoot (with less than perfect syringes), but they will colonize and fruit directly.  From these, I've spore printed the fruits, and will mix them with the original spore prints, make a new syringe, etc.

If you're looking for interesting traits, the only way to do it is produce tons of offspring and start selecting.  The plan was to do run after run, mixing the parent generation's spores with the offsprings, and running it again.  Just through probability, at some point, the genetics should arrive at some sort of genetic "average".


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InvisibleMateja
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Re: Questions about vermiculite and contamination resistance [Re: bodhisatta]
    #26964945 - 10/01/20 10:48 PM (3 years, 3 months ago)

Quote:

bodhisatta said:
I doubt it has anything to do with the ingredients but rather the construction and texture.



:whathesaid:
The structure of a BRF mixture would in technical terms be described as having high 'osmolarity' and being low on the 'water activity' gradient (aw). Osmolarity and water activity are the key principles which will decide what kind of contamination you might encounter in a given substrate and how active the microbial growth will be. I've recently downloaded a bunch of PDF research documents on this and I find it a captivating read to say the least. Here are some screen shots from my phone to give you an idea of what it's about and to steer you in the right direction in case you're interested in learning more about how shit contaminates and why.




And its important to disninguish between water content and water activity which are two separate things.



--------------------
Cakes inside Water Tub


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OfflineJohn in WI
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Re: Questions about vermiculite and contamination resistance [Re: Mateja]
    #26965343 - 10/02/20 06:25 AM (3 years, 3 months ago)

Oh wow.  Thanks for the information.  It turns out I'm a chemist by training.  I'm familiar with osmolarity.  My studies were in physical chemistry, and honestly I never studied anything biological at the university.  Trying to make the leap from p-chem to mycology.

I really appreciate the PDF.  I'll read it when I get back to a computer.


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