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PC'ed Mycelium Survives! wait, that's trich !


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#21 bear

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Posted 14 January 2008 - 12:12 AM

And if anyone can properly test it..it's faht. Once I see his post (who knows if it will become a tek?), I'll consider the matter thoroughly settled

#22 Lazlo

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Posted 14 January 2008 - 12:24 AM

Whether the mycelium is dry or not, it can't withstand those temperatures for any amount of time. I'm guessing your culture is a product of contamination, even if it is from a species of mushroom.

I've cooked many jars of new seed that had residue of mycelium plastered to the jar's walls and not once have even come close to having it thrive on. The culture in that jar is a product of contamination.

#23 fahtster

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Posted 14 January 2008 - 12:37 AM

I am also a skeptic. lol how long was that PC'd for? I'm definitely going to see what that jar looks like in a couple days before I go PC'ing any of my masters. lol it would be an old strain that I don't use anymore anyhow. I have a few of those jars laying around. but I tend to agree with laz on this.

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#24 vinz

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Posted 14 January 2008 - 01:05 AM

skeptic here too but this is really interesting lol
if it is mush myc, it could possibly come out weak due to the stress its been through..

#25 warriorsoul

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Posted 14 January 2008 - 01:08 AM

So youve tried this before Lazlo?
how long did the myc dry for?
how many days after pc'ing before you gave up ?

what you said about a contaminate makes sense
but its worth exploring the limits of dry myc
if not for the simple reason no one really has

i read that freeze and drought tolerance is related in myc
some mycs ability to go into a dormant state when dry
allows it to survive freezing temps.
maybe the same holds true for heat.

i have a digital pc and im not sure what the max psi is
so i dont know what the temp was in my pc.
my contam rates in general have been pretty low
i pc'ed for 60 min
the myc had been sitting for at least a week
with lots of air to dry it out

#26 Lazlo

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Posted 14 January 2008 - 01:46 AM

I haven't tried it, but i've done it inadvertently. I don't wash my jars every time I use them unless i've had contamination in them. Well I do wash them, but every other time or so. Quite a bit there's left over mycelium in them that's visible but doesn't pose a harm when it comes to cross contaminating my new culture, new spawning work.

Although mushroom mycelium can withstand temperatures of 170-200 degrees for a short amount of time. A few seconds. The hot agar pour when doing petri work proves that.

#27 sooshane

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Posted 14 January 2008 - 04:58 AM

Hi,
Thermal death for most Mycelium starts at about 106f. Most endospores, bacteria, etc are killed at the temp of 212f. Pasteurization happens at temps between 140f and 180f and kills many molds and spores - leaving some bacteria alive. Some dry yeasts can survive temps of 250-260f for about a whole 5 minutes. So, even if your PC didn't rise above the 106f mark then any endospores or bacteria present would have also survived and the grain would have contaminated.
So...
Edited to add that I just noticed Invitros post(#4) saying the same thing. Thats what I get for skimming.

#28 Hippie3

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Posted 14 January 2008 - 09:13 AM

i'm keeping an open mind until it's tried
but yeah, i've recycled 'dirty' jars with dried mycelial fragments
and never got growth.

#29 kocos

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Posted 14 January 2008 - 09:22 AM

Same here, ive given lots of opertuneties for this to happen, due to my sloppyness, however ive never seen anything like this before. If you think about it logically, i would doubt that dried mycelium could withstand even boiling temperatures for a short period of time. Not to mention above boiling temps for over an hour.

#30 warriorsoul

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Posted 14 January 2008 - 12:41 PM

i just found a couple specs of contam..
although the myc is myc
the test is flawed now

if contam got in
spores could have also.
this particular jar didnt have a filter
just a nail hole with saran wrap.




thanks:bow:

#31 Lazlo

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Posted 14 January 2008 - 12:51 PM

Hi,
Thermal death for most Mycelium starts at about 106f.


For extended time it is. Every species of mushroom that i've needed to clean up on agar gets a layer of nearly boiling agar poured on top of it to pasteurize the bacteria. Then the mushroom mycelium colonizes up to the top of the new layer of agar. And this hot agar layer stays steamy for around a minute or 2 (you can't put the dishes lid on until the steam has subsided). Mold and mushroom mycelium is a bit tougher when it comes to heat than you'd think. Heat in short amounts of time.

#32 Hippie3

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Posted 14 January 2008 - 12:57 PM

i just found a couple specs of contam..
although the myc is myc
the test is flawed now

if contam got in
spores could have also.
this particular jar didnt have a filter
just a nail hole with saran wrap.




thanks:bow:

one last pic
of the contams
plz
:bow:

#33 CoyoteMesc

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Posted 14 January 2008 - 04:19 PM

For extended time it is. Every species of mushroom that i've needed to clean up on agar gets a layer of nearly boiling agar poured on top of it to pasteurize the bacteria. Then the mushroom mycelium colonizes up to the top of the new layer of agar. And this hot agar layer stays steamy for around a minute or 2 (you can't put the dishes lid on until the steam has subsided). Mold and mushroom mycelium is a bit tougher when it comes to heat than you'd think. Heat in short amounts of time.


I have never read this anywhere before. This is the first. I did however think of trying it after this thread. Agar is hit and miss around these parts. Can you link me a thread or start one soon on the steps you take to do this Laz, Thanks
CM

#34 Hippie3

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Posted 14 January 2008 - 04:22 PM

Mycotopia Web Archive: pouring hot agar-over contaminated dishes

#35 CoyoteMesc

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Posted 14 January 2008 - 04:36 PM

Thanks Hip :bow:

#36 sooshane

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Posted 14 January 2008 - 07:28 PM

For extended time it is. Every species of mushroom that i've needed to clean up on agar gets a layer of nearly boiling agar poured on top of it to pasteurize the bacteria. Then the mushroom mycelium colonizes up to the top of the new layer of agar. And this hot agar layer stays steamy for around a minute or 2 (you can't put the dishes lid on until the steam has subsided). Mold and mushroom mycelium is a bit tougher when it comes to heat than you'd think. Heat in short amounts of time.


I don't understand the use of pasteurized agar or exactly your method that you describe here. This almost sounds like bottom inoculation wherein a piece of tissue is put in the plate under the poured and cooled agar. Are you pouring this over Mycelium or a live tissue sample from a fruitbody? I'm thinking that if a tissue sample is large enough that it could survive several minutes at sub-boiling temps. But, I don't understand the reasoning behind pouring hot agar onto tissue.

#37 CoyoteMesc

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Posted 14 January 2008 - 08:59 PM

Well I cant speak for Laz, but here is my thinking.

We all know that Agar it highly acceptable to airborne contams when cooled.(usually when we do our transfers) But if you were to do this while the agar is still hot, well then your mycel. should have a far better success rate. Since the lid could go right on. Mycel and agar in the plate with the lid. When the agar comes down to temp where its acceptable to contams, well we wont care cause its all closed up.

Another reason would be whats covered in that link that Hip posted. If you got a plate with some nasties and some healthy mycel. you can pour the hot agar over the old layer(covering mycel. and nasties) and the mycel will colonize through to the top.

#38 Hippie3

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Posted 14 January 2008 - 09:06 PM

the hot molten agar kills off many lesser lifeforms
but the mycelia survives,
thus reducing contam levels

#39 CoyoteMesc

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Posted 14 January 2008 - 09:08 PM

:lol:

yeah what he said,

lol, you could prob guess I got kids considering how I try to explain everything out...lol

#40 sooshane

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Posted 14 January 2008 - 09:28 PM

Thanks Hip Coyote,


That's interesting and I understand the process and idea now. However, I do have some questions rolling around in my head. Like how does the Cubensis Mycelium survive? Is Staments wrong again in saying that the thermal death is 106f for cubensis? If you see a contamination like mold then most likely by that point it has already given off it's spores. Pasteurization is about 99.9% efficient. The thermal death points for most bacteria, molds and spores during pasteurization are as follows, 212F - to 185 F, when heated for a few seconds; 180 F for 2 minutes; 170 F for 5 minutes; 140 F for 20 minutes. Even some thermolphilic fungi and it's spores can only survive up to about 145f.


Seems like it would just be easier to section and isolate but I have some colonized plates that I think will get some heat play.

Question for Lazlo. What temp (approx) are you pouring at?




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