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10-12 year old spore print, back from the dead, zombie agar!


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#1 YoshiTrainer

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Posted 11 February 2020 - 06:11 PM

A quick bit of history:
In the waning years of the 20th century, a FOAF brought an unamed cubensis spore print back from Amsterdam. That coupled with an old, used PF TEK kit allowed for many wonderful camping trips and spiritual journeys. As time passed, life got in the way of living and spore prints were stored "for later". 10-12 years later, the spores ended up in my FOAF's, friend's, cousin's, plumber's, friend's possesion. Instead of using FOAFFCPF, I'm just going to use FOAF or he/him from now on.

After being stored in folded card stock taped closed in a dresser drawer, my FOAF prepared a spore syringe like usual but used the entire print. He used it to inject jars of PCed popcorn and was met with massive contamination! Liking a challenge, my friend dropped pieces of the cleanest popcorn kernels onto agar.

From that nasty agar, transfered a chunk to new agar, from that, a smaller chunk to new agar. From that, start over again because his box full of jars tipped over and the condensation apparently washed over the remaining contamination, spreading it again. After taking a scraping to new agar, then to new agar he was still fighting Cob Web at the very least.

After transferring a scraping to new agar, he added some H2O2 diluted to 5% to the old agar and swirled it around. This was a very bad idea!

The transfered agar grew out for a couple days again and then a scraping was transferred. The new old agar was then subjected to a 5 minutes soak of H2O2 diluted to 10%. After the H2O2 was dumped out all of the infected mycelium except it's very outer ring was cut out and removed. I think next time, he should do that in reverse, cut out then soak in H2O2. He also started over from a 2nd spore print and only scraped a tiny bit from the middle of the print, first transfers were done today.

The hope is to get rid of the contamination or knock it back so far that the mycelium can fight back or out grow it. At this point though, it is frustrating, he is stubborn and mad now, so the fight is on! He says it smells of cubensis mycelium, just soured so there is hope.
Any tips I can pass on to him would be helpful.

It would be incredible to see these mushrooms fruiting again after 10-12 years of storage!

Thank you!

1st pic spore print
2nd folded print
3rd corn on agar
4th 1st transfer
5th 2 transfers after starting over
6th week after 5% H2O2
7th same
8th same

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Edited by YoshiTrainer, 11 February 2020 - 07:13 PM.

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#2 Fungi2b

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Posted 11 February 2020 - 06:26 PM

I don't wanna be a "Debbie downer" but that agar all looks shot to shit buddy.
Anything is possible tho, I was recently reading that someone took Amber that had fruit bodies of a prehistoric fungus trapped in it and cut it open and eventually got it to grow. It was a he'll of a process involving a lot of transfers and a lot of fancy lab equipment. But that's what gave me the idea to try resurrecting all my old samples.
I wish you the best of luck but wouldn't put or open that agar anyplace near my grows.
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#3 Misfit

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Posted 11 February 2020 - 06:37 PM

I wish you the best of luck but wouldn't put or open that agar anyplace near my grows.

Came to say this
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#4 YoshiTrainer

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Posted 11 February 2020 - 07:45 PM

Thank you friends! The fifth photo is more what he is currently working with. It is still F/up but not like the last 3 photos! I know he loves a challenge and already has looked into peroxide/iodine, garlic, baking soda, maybe tea tree oil? :)

Edited by YoshiTrainer, 11 February 2020 - 08:01 PM.


#5 Ripples

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Posted 11 February 2020 - 08:10 PM

I think you're just transferring mold from one to another. I don't see anything in those agar dishes that resembles Cube mycelium. Just because its white and you have isolated away from the bacteria  doesn't mean its what you're after. I do hope I'm wrong but I hate to see you get your hopes up based on what you've shown.


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#6 Fungi2b

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Posted 11 February 2020 - 09:06 PM

I'd def just throw it all away buddy. I agree that you guys are just transferring mold and isolating the strongest mold.
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#7 Myc

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Posted 11 February 2020 - 09:30 PM

Every one of those plates are contaminated beyond hope. Unless you're growing mold on purpose.


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#8 macgyver

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Posted 11 February 2020 - 09:42 PM

Worth a try :P


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#9 Fungi2b

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Posted 11 February 2020 - 09:52 PM

This is what cube myc will look like on agar plates.15102858793742000258968.jpg.2cc3e9ba89093b90f64fef50c0f3abad.jpg 15102859325181064860243.jpg.9c245191ace604bcd2fcf25cabc36442.jpg
And this is trich myc1506827763550-1168253656.jpg.59f066171642c8fc9a7fa6ba395beff7.jpg
Notice how the cube myc has that fanning pattern and the trich don't? Those fans are all sub strains that can be isolated. The trich tends to grow as a "blob" without the fan lines (rizos).
My cube samples don't show many different fans meaning I was getting it pretty well isolated.
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#10 macgyver

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Posted 11 February 2020 - 09:57 PM

Good myc on a sub looks greyish white, when trich pops up it tends to grow on top of the myc and almost bubbles, it grows fast and it is starkly brighter than the myc. Very easy to spot once you know what you are looking for, and if you check often, but also very easy to miss if you don't look for a couple days... I've never seen it on agar before!


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#11 YoshiTrainer

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Posted 11 February 2020 - 11:37 PM

Thank you all for responding and sharing your great information and photos. I agree, the agar jars are very nasty, contaminated and might be hopeless! That being said, there is mycelium in there with all the muck. They smell like mushroom and he's been doing test runs on small jars of grain that partially colonize before infection takes over. At this stage, with no other projects like this, he has nothing to loose and everything to gain. Plus, agar work is oddly fun. :)

#12 Fungi2b

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Posted 12 February 2020 - 12:30 AM

I love agar twerk myself, there is something to it that makes me feel oddly godlike.
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#13 YoshiTrainer

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Posted 12 February 2020 - 01:37 AM

Ha ha, I've never thought of it that way but...yes, I get it.

#14 YoshiTrainer

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Posted 12 February 2020 - 07:18 PM

Today, some tests on old contaminated agar. There were 5 minute soaks of 5%, 10%, 25% and 50% H2O2/iodine solution followed by a sterile water rinse. Lotrimin antifungal cream was also applied to othet jars in 100%, 50%, 25% and 5% (ish) concentrations. We shall see what the next few days hold.

#15 YoshiTrainer

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Posted 13 February 2020 - 01:10 PM

Still fighting!
There were 4 pint jars on chicken scratch fully colonized with some issues. Two were crumbled into a tray and cased with verm. The other two had some contamination on the bottom so, the bottom 1/2 of the cakes were cut off and disposed of. The top 1/2 of the cakes were soaked for 12hrs in a very mild H2O2 solution then rolled and cased with verm. He just needs one mushroom to declare victory, either a print or clone.

Fingers crossed!

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Edited by YoshiTrainer, 13 February 2020 - 01:11 PM.


#16 Fungi2b

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Posted 13 February 2020 - 01:54 PM

Good luck.
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#17 YoshiTrainer

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Posted 14 February 2020 - 03:54 PM

There was an original 10% H2O2 wash and cutting out of the center of the bad stuff to this agar. When that didn't work, the dirty agar was soaked in H2O2/I solution of different concentrations. Others were subjected to Lotrimin cream also at different dilutions. A couple transfer's of the surviving "white stuff" from the higher concentration H2O2/I soaks were done. Mostly to see what survived, if anything.

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#18 smellitstinknot

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Posted 14 February 2020 - 10:26 PM

All the white growth in your pictures is mold mycelium. The leading edge from which you're transferring is white because it hasn't yet sporulated.  It'll turn green every time you make a transfer from those plates.


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#19 YoshiTrainer

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Posted 15 February 2020 - 01:47 AM

Thank you Smellitstinknot, all the agar was pitched. This was just and experiment, There were a couple scrapings from white "stuff" that barely survived the 25% and 50% H2O2/I. It already looks like mold growth but we'll give it one more day. Then all the nasty agar is gone and major clean up begins!

#20 DocOct

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Posted 15 February 2020 - 02:59 AM

If you have mycelium on cakes,... why didn’t you just take samples from them???

All the time/materials spent transferring mould could have been used to expand your current mycelium exponentially.

Next set of agar dishes either should get samples taken from the cakes or samples taken from your cased sub.

Your friend should be rolling in mycelium by now mate.

Edited by DocOct, 15 February 2020 - 03:03 AM.





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