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Non-sterile experiment

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#1 newmoon



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Posted 26 February 2019 - 01:18 AM

Hello all,


After a long time of reading this site but having nothing to share, here's something new. This experiment is still in its early stages, but perhaps it will be of interest. Any replication or refinement is much appreciated.


The goal is to find a completely non-sterile method of cultivating mushrooms, based on a couple of observations:

1. Established mycelial growth is resistant to contamination

2. Some substrates are resistant to contamination, but won't directly allow germination.


The idea, then, is to germinate the fungi on a minimal amount of material, and then transfer to a resistant substrate and build up mass gradually from there.


Materials used:

1. Oatmeal - "old fashioned rolled oats"

2. Coir

3. Worm castings

4. Small capped test tubes (15ml or less) or tiny jars.

5. P. cubensis "B+" spore syringe from a commercial vendor


Nothing is sterilized or pasteurized.


Place one oat at the bottom of each tube, dry. Add one drop of liquid from the syringe (this is enough to hydrate the oat). Seal the tube and let sit at room temperature. Once the oat is mostly colonized, add a small amount of 50/50 coir/castings mix on top. Once that's colonized, go from there. Alternately, transfer the colonized oat to damp cardboard and expand that way.


After a week, 14/20 tubes have shown growth, 4/20 have grown mold, and 2/20 have grown bacteria. A picture is attached of one tube starting to colonize the "bulk" substrate. More replicates will be performed and better pictures added as time allows.


This also appears to work with Psilocybe cyanescens, transferring to cardboard after the oat is colonized.


#2 Myc


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Posted 26 February 2019 - 09:51 AM

Welcome newmoon!

#3 newmoon



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Posted 26 February 2019 - 10:21 AM

Thanks, Myc! I've learned a ton from your posts over the years, especially about wood lovers.


Here's a picture of an oat that was transferred to cardboard two days ago. Toilet paper roll moistened with reverse osmosis water. This one is also B+ cubensis.



#4 Microbe


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Posted 26 February 2019 - 07:21 PM

I love your thought process. The growth on the cardboard, looks damn good. I love experiments and will surely be checking in on this frequently. Im about to resume all my experiments i had to place on hold because of life and cold ass weather!

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#5 newmoon



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Posted 28 February 2019 - 12:33 PM

Thanks, Microbe!


The tube in the first picture (B+ cubensis) has been coming along nicely, so today I transferred it to a 1/2 pint mason jar and added a layer of coir/castings on top. Hoping that will colonize well.



The B+ cubensis oat on cardboard has also expanded well. I imagine one could do non-sterile strain isolation and other agar techniques on cardboard like this (I know that other people have experimented with this before), but I'm not going to bother this time around. So, I trimmed the cardboard, gave it a day to keep growing, and then put it in a jar with coir/castings.



Finally, here's an oat colonized by Psilocybe cyanescens deciding whether it wants to move to cardboard. This is also non-sterile, I just put it in a petri dish with a parafilm wrap because it was a handy container.


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#6 newmoon



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Posted 03 March 2019 - 03:36 PM

Unsurprisingly, the "bulk" jar with the first B+ tube started growing mold. I did some surgery to see if I can recover anything, but I'm not optimistic. More where that came from, anyway.


This did make me realize that I'm still thinking too much in the sterile way, where one grows inside a jar with a few air holes covered in micropore tape. This doesn't work here, obviously. So, in the future I'm going to try making it closer to an open air situation after the initial piece of grain is colonized.


I've also started an experiment to get a better understanding of the germination rates with this approach.


tubes - Copy.JPG

#7 newmoon



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Posted 12 March 2019 - 11:15 AM

I feel pretty comfortable saying that starting on an oat and then transferring to cardboard is a viable non-sterile first step for both ps. cubensis and ps. cyanescens. Somewhere around 1/3 of the oats get to this point, after which I transfer to cardboard:



I think the tubes are still too large, and so moisture control is difficult. I'll be trying some more oats in much smaller containers, and I expect that the success rate will be higher. Even with 1/3 success, I think it's ok. The cost is pretty much free and it doesn't require any time to set up.


Ps. cyanescens, just moved to a bigger jar of damp cardboard:

cy.JPG . I plan to expand this out on cardboard/paper/wood chips.


Ps. cubensis after a few days on cardboard:

cube.JPG cube2.JPG


I'm still playing around with different ideas for the next step for cubensis. I'll also be starting some other wood lovers and oysters to see how those work.

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#8 Sumgi



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Posted 12 March 2019 - 12:26 PM

That's looking great, thanks for keeping us informed! 

#9 newmoon



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Posted 20 March 2019 - 12:49 PM

The coir/castings mix seems prone to mold in sealed containers, so next I'm going to try castings alone. I've kept castings in airtight containers for storage for many months with no sign of growth.


Took this oat on cardboard:

growth - Copy.png


Cut it approximately as the sectoring suggested:

cut - Copy.png


Put each piece in a jar of castings:

jars - Copy.png

Each lid has a 9/32" hole, with a single layer of micropore tape to keep humidity in and insects out. The castings have been stored in freezing conditions to hopefully kill off any invertebrates.

#10 newmoon



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Posted Today, 02:25 PM

I've started a good number of jars like the one in the previous post. Here's the one with the most growth:


The hyphae are moving through the castings at a reasonable speed, but the growth isn't dense - just tendrils. There are no obvious signs of other molds competing. Does anyone have an interpretation of this? I'm wondering if it could be a lag time switching from eating cardboard to eating castings, but I don't know.

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