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Will they germinate? The Old Spores Thread...


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#41 Freaky

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Posted 20 October 2020 - 10:27 PM

It's only partly about this topic.

What about the storing of exotic spores and edible mushs ?
Is it best for them to be stored in the fridge too or even frozen ?

I remember the info from david barlow that copelandia cyanescens are best stored in the freezer but thats the only time I've heard about freeze storing of spores.

I'll get Hericium and versicolor very soon and want to start agrocybe aegerita aswell, not sure how to store their spores and my exotic spore library to keep them intact as long as possible

 

 

I wonder that also - shelf life of exotic and edibles is definitely worth discussion.  I am sending out some exotic prints mostly Pans and hope we can see if they come back or if there is a best practice for storage. Someone else mentioned they had luck with frozen culture that came back.  Maybe DickMoby????

 

It's been a while since I read through my copy of TMC but I may see if there was any detail about long term storage beside cold and dark or sealed in the fridge. 



#42 DickMoby

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Posted 21 October 2020 - 06:40 AM

r3zwyb27.jpg

 

Those are the repeatedly frozen cultures.

As you can see on the bottom right picture, I ripped some of the rubbery and partly dried myc from outside of the cut area with it, it was nothing like living mycelium.

 

o833ztpd.jpg

 

Here is the Rusty Whyte plate that didn't want to germinate over three to four months.

It's the only plate under them with viable mycelium, also after the accidentaly repeated freezing periods it finally germinated.

 

Here a picture of all plates together

 

ib9anye4.jpg

 

Upper left corner is the Rusty Whyte plate.

All the other plates were living mycelium that was frozen, while that plate was inoculated with spores that didn't want to germinate.

 

Here some of the plates after they've been transferred to a different agar recipe (the transfer pieces were placed mycelium down on the new plates.

 

dltdmuy3.jpg

 

The green is Spirulina.

It took a while until they recovered but now they're growing back.

Kape just looks that weird because the myc is wet from the no pour excess water in the plate, it will grow out and show normal growth in a bit.

 

The growth overall is slow and I can't wait to fruit them to collect spores and start from new, those plates are my only sources for those gens.

_____________________

 

Those were living cultures tho and I DON'T recommend to freeze cultures as storing method! 

That was accidentally and just a rescue mission.

 

Really wonder if freezing spores is a generally good method for long term spore storage


Edited by DickMoby, 21 October 2020 - 06:42 AM.

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#43 Freaky

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Posted 22 October 2020 - 04:44 PM

Thanks for sharing that on the frozen plate.  Definitely opens the door for further science to be done for comparison. 


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#44 DickMoby

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Posted 23 October 2020 - 12:29 PM

Yea, definately gonna give it a try.

The smaller plates came today

 

IMG_20201023_191001.jpg

 

I'm ready.

 

Here some agar from Radical mycology

 

PSX_20201023_191827.jpg

 

The first ones are spore germination recipes, could be interesting for here


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#45 DrepsiLocybe

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Posted 24 October 2020 - 06:47 AM

Yea, definately gonna give it a try.
The smaller plates came today

attachicon.gifIMG_20201023_191001.jpg

I'm ready.

Here some agar from Radical mycology

attachicon.gifPSX_20201023_191827.jpg

The first ones are spore germination recipes, could be interesting for here

those plates are nice, ive been using glass dishes for a couple of years now.

my advice is to dry sterilize them (wrap them on foil and bake them @ 350f for 1 hour).
and dont over stack them when pouring.
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#46 DickMoby

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Posted 24 October 2020 - 07:14 AM

"my advice is to dry sterilize them (wrap them on foil and bake them @ 350f for 1 hour).

and dont over stack them when pouring."

 

 

Cheers!

I'm using glass plates since the beginning, had some micro boxes with airfilters as large plates in between but that just to have one huge plate with a single inoculation.

 

All my plates are no pour sterilized with the media.

Its a pain to pour plates in the SAB, with no pour I don't have a single contam in my plates so thats for me the way to go until I upgraded.

 

I'll give the oven a try once I built a flow hood, right now tho the money is needed at other ends.

Definately gonna switch to pour with the flow hood!

 

:thumbs_up:


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#47 DetritusTheEgo

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Posted 24 October 2020 - 08:37 AM

All my plates are no pour sterilized with the media.

Its a pain to pour plates in the SAB, with no pour I don't have a single contam in my plates so thats for me the way to go until I upgraded.

I'm thinking about taking the plunge and buying some glass petris. I already do no pour PP5 containers for agar since I as well don't have a flow hood setup and have found it a PITA to time the agar cooling period as well as pouring in general in a SAB.

 

What's your no pour glass petri method look like? I assume you stack the petris inside a vessel of some kind or elevate the petris above the water line with the canning rack, an empty row of half pint or pint jars, and an additional canning rack for a flat surface? If it's quite off topic I can IM or possibly inquire in another thread :biggrin:


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#48 Freaky

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Posted 24 October 2020 - 12:26 PM

Yea, definately gonna give it a try.

The smaller plates came today

 

attachicon.gifIMG_20201023_191001.jpg

 

I'm ready.

 

Here some agar from Radical mycology

 

attachicon.gifPSX_20201023_191827.jpg

 

The first ones are spore germination recipes, could be interesting for here

I'll be getting some agar work done today - some no pour in PC and I may attempt some petri pour in the SAB but like  you I hate using the SAB for agar work.  Tried a trial run the other night and it did not go well so I'm debating trying to pour in there again. 

 

Thanks for those recipes - I have malt extract syrup - I prefer it over powered/dry malt extract - picking up some yeast today and going to try the activated coconut charcoal additive also.  I'll be sure to post the breakdown of the agar recipe's. 

 

 

 

 

All my plates are no pour sterilized with the media.

Its a pain to pour plates in the SAB, with no pour I don't have a single contam in my plates so thats for me the way to go until I upgraded.

I'm thinking about taking the plunge and buying some glass petris. I already do no pour PP5 containers for agar since I as well don't have a flow hood setup and have found it a PITA to time the agar cooling period as well as pouring in general in a SAB.

 

What's your no pour glass petri method look like? I assume you stack the petris inside a vessel of some kind or elevate the petris above the water line with the canning rack, an empty row of half pint or pint jars, and an additional canning rack for a flat surface? If it's quite off topic I can IM or possibly inquire in another thread :biggrin:

 

 

I may try some PP5 deli containers - can order the clear ones on Amazon in bulk so I may give those a try.  

 

I encourage everyone to share - please feel free.  That's what this thread is for really - everyone to discuss and maybe we will stumble upon something that helps on old prints in the library or concedes that they're just useless and dead after so much time spent on foil.


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#49 Freaky

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Posted 24 October 2020 - 12:28 PM

Yes, and I'm going to build a flow hood - I know I wasn't planning on it, but I want to really dig into agar and work with these old prints and a flow hood is going to make that work much cleaner and more efficient. 

 

I only need to buy the filter and my friend is a carpenter so he can do the construction lol. 


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#50 DickMoby

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Posted 24 October 2020 - 01:48 PM

Gonna post it here because we are also talking about agar.

 

I just fill them with the media and let them cool for a couple mins to solidify

 

inxqq7ir.jpg

 

Once hardened, they are packed in tinfoil

 

hgxlc3ul.jpg

 

For agar and LCs I use a small pc that came with this thing idk if its for steaming vegetables or something else, it does the job perfect to make enough space for the water and has a flat bottom.

Packed some of the small plates into jars to see if they are less wet, helped a bit.

 

Short note before the other pictures, I don't work with those gloves in the SAB, was just for the pictures, didn't want to waste a single use glove for the pics.

 

me6bfmkc.jpg

 

The bottom plate is pulled up to close the gap on the upper side

 

ldgjhqaj.jpg

 

and to open the gap on the lower side

 

xo4lhv57.jpg

 

then turned upside down and tapped twice on the clean paper towel to collect most of the water in the lid 

 

abvkqd6e.jpg

 

after the tap its turned back to normal to tap it again twice on the paper towel to get the water out through the gap.

I wipe them dry after that and inoculate.

 

Due to the amount of water that needs to be collected with paper towel, I do max 12 plates at once, otherwise there is alot of water in the SAB.

 

No idea if thats understandable, I'm doing it like that for a good while and have no contam issues.

There will still be some moisture left but its not a big deal, incubated the myc uses it up pretty quickly (on room temp not so fast).

 

Pouring is not fun in the SAB, this process here is ok, I mean routine really makes it a quick process, if someone has a really large SAB like 120qt then its possible to do more plates but in my 80qt SAB with the iso bottle, torch and the transfer plates or prints and I use a peroxide soaked paper towel to cool the flame sterilized scalpel instantly + an iso soaked paper towel (to wipe the old plates) and the towel to dry the fresh plates, space is pretty limited.

 

A flow hood is badass compared to that, we don't need to talk about that but not necessary at all to get clean cultures and transfers


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#51 Freaky

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Posted 24 October 2020 - 04:16 PM

Thanks for sharing that DickMoby!

 

I got sidetracked today doing some other things so haven't started my agar cook yet lol

 

My SAB is 76 QT I think and yes, it's a tight fit but I'm hoping a trial pour to those ketchup cups will be fairly easy since it's all on a smaller scale to pour them.  I'll probably make a mess and have agar all over the SAB.


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#52 DrepsiLocybe

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Posted 24 October 2020 - 04:45 PM

"my advice is to dry sterilize them (wrap them on foil and bake them @ 350f for 1 hour).
and dont over stack them when pouring."


Cheers!
I'm using glass plates since the beginning, had some micro boxes with airfilters as large plates in between but that just to have one huge plate with a single inoculation.

All my plates are no pour sterilized with the media.
Its a pain to pour plates in the SAB, with no pour I don't have a single contam in my plates so thats for me the way to go until I upgraded.

I'll give the oven a try once I built a flow hood, right now tho the money is needed at other ends.
Definately gonna switch to pour with the flow hood!

:

nice, i will give your method a try :).
last time i tried the no pour it spilled a bit and was a pita to clean but i didnt do it like you do.
i just literally pour them and then stack them up and pc them.
ill try your method next time i make a small batch.
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#53 mushit

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Posted 24 October 2020 - 05:25 PM

My SAB is 76 QT I think and yes, it's a tight fit but I'm hoping a trial pour to those ketchup cups will be fairly easy since it's all on a smaller scale to pour them.  I'll probably make a mess and have agar all over the SAB.

Ha ha ha.  Been there.  Done that.

 

As far as flow hoods go, I am having a hell of a time trying to find a proper filter.  They are not available to the consumer, and when I call a commercial supplier, they don't even want to talk to you.

 

Guess I will be using my SAB for a while....

 

edit: Oh, and I also like the glass dishes.  Just wash them, dry them, wrap them in foil and bake.  Good for the environment as well. 


Edited by mushit, 24 October 2020 - 05:31 PM.

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#54 clumsy

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Posted 24 October 2020 - 07:20 PM

As far as flow hoods go, I am having a hell of a time trying to find a proper filter.  They are not available to the consumer, and when I call a commercial supplier, they don't even want to talk to you.

 

I made a HEPA air source here. I had to order two filters but only needed one. I will part with it for $100.00 plus shipping, which is probably another $100.

 

 


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#55 DickMoby

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Posted 25 October 2020 - 06:56 AM

Thanks for sharing that DickMoby!

 

I got sidetracked today doing some other things so haven't started my agar cook yet lol

 

My SAB is 76 QT I think and yes, it's a tight fit but I'm hoping a trial pour to those ketchup cups will be fairly easy since it's all on a smaller scale to pour them.  I'll probably make a mess and have agar all over the SAB.

 

Same here, sterilized plates but couldn't get to inoc them.

 

I thought about the mess.

 

PSX_20201025_121135.jpg

 

The media bottles have this plastic thing, is it called drip tip ? Idk.. 

Anyways its there to prevent the liquid from running down the bottle and also lets you pour the media straight down without any gravity issues that lead to media spread all over the place.

 

IMG_20201025_120927.jpg

 

To make the pouring easier the plates or cups could stand on something clean to make some room around and raise them from the bottom to have less distance to the containers.

The containers could be closed after the first floor is poured and then just put the next row on top to pour them.

 

I won't try that because I'm strictly anti pour in the SAB lol but it could make your life a little easier to put them onto something.

 

"i just literally pour them and then stack them up and pc them.

ill try your method next time i make a small batch."

 

 

Yea the cooling allows easy handling.

Some other things that prevent headache are to fill the plates under half way to make sure they don't boil over and to also let the pc cool down fully before moving the plates out, takes 4-5 hours of waiting here to be sure that the plates are hard.

_______________

 

To add something to the original topic.

I'm not going to do swabs with the old spores or to only put a few spores on the plates.

Even fresh swabs are sometimes a pain in the butt to germinate, definately gonna throw a mountain of spores on the plates to hopefully have a few of them germinate or syringes with very little water and alot of spores to have a high concentration in the drops that are put on the plates.

 

Pre hydrated (with the rubber band syringe method) spores in a little water put on cornmeal/yeast/charcoal agar seems like a good idea.

 

All the spore germination agar recipes from radical mycology contain cornmeal and yeast seems to be good too for that purpose, with coal as additional germ promoter.

 

Can't wait to start


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#56 mushit

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Posted 25 October 2020 - 11:15 AM

Thanks so much, clumsy! :smile:

I wish I had read this earlier.  I just found a supplier and ordered some.

They are MERV 16 which is the highest rating we can get in the great white north.

Thanks again.

Good bye SAB.



#57 mushit

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Posted 25 October 2020 - 11:21 AM

All the spore germination agar recipes from radical mycology contain cornmeal and yeast seems to be good too for that purpose, with coal as additional germ promoter.

I have never heard of using coal, a petroleum product, in Mycology.

You don't mean charcoal, do you?


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#58 Freaky

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Posted 25 October 2020 - 11:50 AM

Coal/charcoal activated - I used powered coconut charcoal in my agar recipe yesterday.

I’m definitely out of practice with my agar - it’s been 15 years since I cooked any agar and I forgot to add yeast to these first 30 no pour jars. The agar turned out perfect though I just forgot yeast in this round. 500ml poured 24 half pint wide mouth jars then I quartered it and poured six more.

Per 500ml water - 8g agar, 8g malt extract (thick syrup), 1g activated charcoal, small amount (eyeball) brown rice water from boil of brown rice for sub jars. Forgot the yeast but next agar cook will include it. It’s not like I don’t have enough spores to try different agar recipes. Those radical ones I’m definitely going to try out.

I may stay no pour until flow hood is ready. If I do attempt a SAB pour we can all laugh at my spills in the box lol



9E12C92B-319F-42CF-B842-72DEEE9AF948.jpeg
I found some spore syringes from vendors in a separate storage - not kept in a fridge I just stored them in their ziplocks stuffed in a backpack. These are H3 from ralphsters and I have 8 sporeworks syringes. So I figured I would drop some of these to the agar and see what happens. I’m really hoping they’re viable then I’ll just drop into LC and box some jars. But we will see if they grow at all.

Today I’ll be working the spores from 2006 - I have chosen Hillbilly, Acadian Coast(my own) and Malabar.
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#59 DickMoby

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Posted 25 October 2020 - 12:27 PM

Yea activated charcoal, for reasons of lazyness I typed coal, didn't pay out tho beacuse of the extra words for this comment (:

 

The book info says 10g charcoal per 500ml, guess it won't show any beneficial effects with 1g in it.

 

Good luck for the jars!

 

You had PE6 in your list, I've grown them multiple times hoping to get a PE like pheno because its advertised as variety that produces both - texas like and PE like mushs.

Because I can't get PE that was the plan to finally get my hands on some dicks,

ah I can't type it like that, scratch that.. - to finally get a PE.

 

For me and other people aswell it never dropped a single PE like fruit, the genetic was badass tho! Very fast germination and agressive colonization.

 

Did your old PE6 drop both types ?


Edited by DickMoby, 25 October 2020 - 04:35 PM.

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#60 Freaky

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Posted 25 October 2020 - 02:48 PM

I never grew PE6 or Peg I just have a print in the library. So if I figure out a good process and recipe for these old prints I’ll definitely mess with the Pe6 and see what we can get out of it.


Also - I may hold off on spore to agar for today. I’m shit at SAB work and it frustrates me so I will hydrate some syringes from the prints and once flow hood is ready I’ll work on the print to agar again.

An example of SAB errors - dropped the whole acacdian coast print foil into the agar so now have a plate jar with the whole print. Oops
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