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OfflineVibe_Enthusiast
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PC'd Grain - Life Span?
    #25843606 - 02/28/19 03:05 PM (2 months, 23 days ago)

I took the initiative of prepping all of my grain jars because I had time to do so this weekend. But I really didn't think of the full picture because I still am growing out my agar plate that I'm going to drop out to 2 or 3 jars and use one for a g2g.. so I'm going to have some unused grain jar(s) for over a month.

Do you guys think they'll be okay? I'm just wondering if they're going to dry up and be no good and I wasted all of that time and grain... meh.


Edited by Vibe_Enthusiast (02/28/19 03:21 PM)


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OfflineShaperDreaming
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Re: PC'd Grain - Life Span? [Re: Vibe_Enthusiast]
    #25843620 - 02/28/19 03:10 PM (2 months, 23 days ago)

I've let them sit for 5-9 days in the past after PC w/o problem.


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Edited by ShaperDreaming (02/28/19 03:11 PM)


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OfflineVibe_Enthusiast
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Re: PC'd Grain - Life Span? [Re: ShaperDreaming]
    #25843642 - 02/28/19 03:20 PM (2 months, 23 days ago)

Well like I said, mines going to be at least 4+ weeks lol. I should have waited. I just didnt think big picture. I just wanted to get it out of the way because I had downtime and prepping grain sucks


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OfflineShaperDreaming
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Re: PC'd Grain - Life Span? [Re: Vibe_Enthusiast]
    #25843650 - 02/28/19 03:22 PM (2 months, 23 days ago)

Honestly, if you're not using the jars between now and then, just let it sit. Do a thurough examination of it before inoculation, and if it looks clean... go for it? I'd mostly be worried about evaporation after that much time and having dry grains, can you put them somewhere air-tight-ish?


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Edited by ShaperDreaming (02/28/19 03:23 PM)


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OfflineVibe_Enthusiast
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Re: PC'd Grain - Life Span? [Re: ShaperDreaming]
    #25843668 - 02/28/19 03:28 PM (2 months, 23 days ago)

That's really my concern is them drying out. I guess I could go buy a tote to store them in. If it means it'd help in this situation


Edited by Vibe_Enthusiast (02/28/19 03:29 PM)


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OfflineShaperDreaming
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Re: PC'd Grain - Life Span? [Re: Vibe_Enthusiast]
    #25843713 - 02/28/19 03:54 PM (2 months, 23 days ago)

Quote:

Vibe_Enthusiast said:
That's really my concern is them drying out. I guess I could go buy a tote to store them in. If it means it'd help in this situation



:shrug:

Only one way to find out? Buy it mono-tub size so you can reuse it later :wink:


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OfflineMildlyInsane
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Re: PC'd Grain - Life Span? [Re: Vibe_Enthusiast]
    #25843726 - 02/28/19 04:01 PM (2 months, 23 days ago)

I just used some jars of WBS that had sat for about a month. They didn't get very far before contaminating, but that could just be from me being a noob when it comes to G2G.

But, I was also just reading some stuff in the thread linked below, about PC sterilization only really providing a "window of opportunity" for mycelium to colonize, before bacterial endospores or other potential surviving organisms gain a foothold.

Sounds risky to use old grain, but I'm still new to this hobby and learning stuff.

https://www.shroomery.org/forums/showflat.php/Number/25840271/fpart/5#25840271

EDIT* I suppose you could always re PC them before use?


Edited by MildlyInsane (02/28/19 04:02 PM)


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Re: PC'd Grain - Life Span? [Re: MildlyInsane]
    #25843743 - 02/28/19 04:06 PM (2 months, 23 days ago)

I've weighed out oats before sterilization and over 30 days after and there was no difference in weight loss. I sterilize my oats for 2 hours at 15-20 PSI and left them covered with foil.


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OfflineShaperDreaming
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Re: PC'd Grain - Life Span? [Re: stareatclouds]
    #25843756 - 02/28/19 04:15 PM (2 months, 23 days ago)

Quote:

stareatclouds said:
I've weighed out oats before sterilization and over 30 days after and there was no difference in weight loss.



So, you're saying you've safely left grains out for 30 days after PC and used them w/o contam or stalling from dryness?


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Edited by ShaperDreaming (02/28/19 04:15 PM)


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Re: PC'd Grain - Life Span? [Re: ShaperDreaming]
    #25843775 - 02/28/19 04:23 PM (2 months, 23 days ago)

They didn't have visible contams on their own. I don't have meaningful results as to their viability as I used iffy cultures on the older jars. Definitely had some jars perform well, while others didn't do so hot, but didn't contam (senesced cultures). I'm not recommending doing this or saying it's fine.

People definitely run grain for 3 hours. You can just do 2 hours and run it again for another hour a month later if you're concerned.

This is one of those topics where everyone repeats the same information without having firsthand experience. People quote the "small window to colonize" but have no science to back whether every endospore is killed or not (and I'm not picking a side either way). Using jars immediately is best practice. There's some leeway in waiting, but too many variables play into how far you can stretch it, and nothing concrete exists, so proceed with caution.

Do it and report back. It's not like your life is over if it doesn't work out.


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OfflineShaperDreaming
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Re: PC'd Grain - Life Span? [Re: stareatclouds]
    #25843780 - 02/28/19 04:26 PM (2 months, 23 days ago)

Quote:

stareatclouds said:
Do it and report back. It's not like your life is over if it doesn't work out.



That's where I'm at with my thoughts.

Also, good to know about 30 days out. I assume with properly built jars this shouldn't be a problem.


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Re: PC'd Grain - Life Span? [Re: ShaperDreaming]
    #25843802 - 02/28/19 04:32 PM (2 months, 23 days ago)

I wouldn't use them personally. You can never get grains 100% sterile, all you can do is pc them to kill enough contaminants to give your myc a fighting chance to colonize. It really is just a race against the clock. I wouldn't waste a good agar wedge on old grains when I could simply run a fresh batch and give myself a chance. Sure, you might get lucky, but I don't like failure


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Re: PC'd Grain - Life Span? [Re: sinisterminister7]
    #25843809 - 02/28/19 04:34 PM (2 months, 23 days ago)

Used jars that I had in the fridge, cling wrapped, for over a month.  No issues that I can remember.


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Re: PC'd Grain - Life Span? [Re: sinisterminister7]
    #25843872 - 02/28/19 04:54 PM (2 months, 23 days ago)

Quote:

sinisterminister7 said:
I wouldn't use them personally. You can never get grains 100% sterile, all you can do is pc them to kill enough contaminants to give your myc a fighting chance to colonize. It really is just a race against the clock. I wouldn't waste a good agar wedge on old grains when I could simply run a fresh batch and give myself a chance. Sure, you might get lucky, but I don't like failure




This is absolutely not correct. Why do you think that? Have you tried it yourself?

I have on several occasions sterilized grain jars, for not more than 1 to 1.5 hours, which have last months (and probably would have last indefinitely) at room temperature. Usually with foil over the lid, and in a plastic bag, otherwise they tend to dry out in my climate.

The sterilization time will depend on the pressure cooker and heat/gas transfer situation, but in my opinion, many people here overdo it.


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Re: PC'd Grain - Life Span? [Re: Pinback]
    #25843899 - 02/28/19 05:09 PM (2 months, 23 days ago)

Quote:

Pinback said:
Quote:

sinisterminister7 said:
I wouldn't use them personally. You can never get grains 100% sterile, all you can do is pc them to kill enough contaminants to give your myc a fighting chance to colonize. It really is just a race against the clock. I wouldn't waste a good agar wedge on old grains when I could simply run a fresh batch and give myself a chance. Sure, you might get lucky, but I don't like failure




This is absolutely not correct. Why do you think that? Have you tried it yourself?

I have on several occasions sterilized grain jars, for not more than 1 to 1.5 hours, which have last months (and probably would have last indefinitely) at room temperature. Usually with foil over the lid, and in a plastic bag, otherwise they tend to dry out in my climate.

The sterilization time will depend on the pressure cooker and heat/gas transfer situation, but in my opinion, many people here overdo it.




It absolutely is correct, apparently.  Mad season just finished setting us straight in  another thread just this morning. Not saying I don’t agree with you on the rest,

Quote:

Mad Season said:
FWIW, IME, grains overridden with endospores aren't easy to work with. The PC reduces a lot of them, but it doesn't kill them all. True sterilization is something like 1 particle per 1000000000? It's really just probabilities. THere's a reason why people throughout shroomery history preach that you have to inoculate grains immediately, and jars after a couple weeks should just be trashed and restarted.

Quote:

Pastywhyte said:
Organic grain often has endospores added as an alternative to pesticide. I have found with such grain that often a single cycle is not enough and that bacteria can recover and become noticable in as little as a few days or as long as a few weeks as two weeks. When I started doing muda style bottles it was strange because you could see the bacteria pop up around some of the grains like little satellites. Became suddenly very easy to see what was happening.

When spawning such grain its my experience that you often cannot smell them, yet they will inhibit recovery enough for molds to move in. Sometimes a very high spawn ratio could help but not always. My solution was a two part sterilization which did the best job.

Also its worth noting that I stopped overnight soaking two years ago.



Quote:

Pastywhyte said:
Quote:

Wydue Wanano said:
This is the first mention I've ever heard of "invisible contams." If an endospore is so damaged it can't reproduce, isn't it essentially dead? What effect do these have on growth if they are not reproducing?




Endospores germinate into bacteria, the bacteria reproduces and some of them become endospores when conditions deteriorate. A damaged endospore is perfectly capable of recovering but it does take time. When we "sterilize" we actually don't kill all the endospores. You would need to run an 8 hour PC cycle to do that. We damage them enough to provide a "window with which to colonize". But there is always a possibility that a couple can recover and start problems. Usually this means poor to no colonization of some grains which are then easy pickings for mold.

Most of the time I doubt people even realize this is happening. But it does. To fully kill all endospores you need to go to pretty extreme lengths. Incidentally this is why GLC is a terrible idea.



Quote:

RogerRabbit said:
'Sterilizing' for 90 minutes gives a window of opportunity to get your grains colonized before surviving organisms can get a foothold.  It's not absolute sterilization, which would take up to 24 hours and ruin the grains.
RR



Quote:

RogerRabbit said:
Quote:

but since a pressure cook at 90 min kills all forms of mold and bacterial




Says who?  My experience shows otherwise.
RR




I mean there's many others, and I've had many conversations with people about this. I still stand by the fact that a dirty ass grain like say popcorn which harbors more endospores is an unnecessary risk.




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Re: PC'd Grain - Life Span? [Re: BeefSupremeJr]
    #25843945 - 02/28/19 05:25 PM (2 months, 23 days ago)

Thanks for your reply, but I disagree with that. I suggest that anyone who doubts this try it.

Sterilization in the microbiological sense means that ALL living organisms are eliminated. It is either sterile or not. No probabilities. There is plenty to read about this in books, academic papers, patents... I have not seen it disputed anywhere really, except here.


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Re: PC'd Grain - Life Span? [Re: Pinback]
    #25844028 - 02/28/19 05:56 PM (2 months, 23 days ago)

Nobody debates with sterilization is. People debate whether our jars are truly sterile. This isn't a debate with an obvious answer that satisfies all parties.

Different grains have different endospore counts and bacteria present. Different prep methods will result in more endospore germination and creation. I haven't seen anything definitive showing x hours at x PSI for x volume of grain kills everything that would be useful for our purposes.

And your jars appearing fine without pulsating green doesn't mean they're sterile or as healthy/viable as they were a few hours after the PC cycle. I'd love to see documented grows with old jars and the tubs they produced.

The optimal play is PC your grains for 90-120 minutes at 15PSI and inoculate as soon as possible with healthy mycelium. If you want to push it beyond that, go for it. It's your grain. Definitely log the relevant metrics and post the results. I'd love to see actual trials with this shit, even though the variance involved ensures our results will be different.


Edited by stareatclouds (02/28/19 06:07 PM)


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Re: PC'd Grain - Life Span? [Re: stareatclouds]
    #25844062 - 02/28/19 06:12 PM (2 months, 23 days ago)

:whathesaid:
No one has logged this in any manner let alone something scientific. Everything is someone's story. The less experience people have the longer it seems they're able to anecdotally let things go and be just fine.


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Re: PC'd Grain - Life Span? [Re: bodhisatta] * 1
    #25844105 - 02/28/19 06:28 PM (2 months, 23 days ago)

This is my story :typing:

Rye jar with SFD 120min 18 PSI about a month later and 4-5shakes later Nothing visible at all. This is a test jar.




I personally truly believe this rye in the jar is sterilized, but I leave room for other 'personal' experiences as well.


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Re: PC'd Grain - Life Span? [Re: bodhisatta]
    #25844106 - 02/28/19 06:29 PM (2 months, 23 days ago)

Wasn’t trying to ruffle your feathers pard.  Just learned this myself. Although not documented, I am sure rr has his reasons for saying it.  The man has literally watched spores germinate in real time under a microscope. 

Ima go with rr’s decades of xp on this one.  I don’t like risks, particularly when unnecessary.


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Re: PC'd Grain - Life Span? [Re: BeefSupremeJr]
    #25844112 - 02/28/19 06:34 PM (2 months, 23 days ago)

Practically no spores and organisms we deal with can  survive 120C for 20 minutes.
The rest of the PC cycle time is spent on making sure this high temp has penetrated to the center of the jar. Let's say only 115C is able to reach into every grain. Still 15min of 115C kills the shit out of the contams we deal with usually. So I'm basically saying if the PC cycle is proper the grain should be sterilized. I make this statement mostly cause it's so unplausable to me to think of a scenario when a proper PC cycle wouldn't kill everything inside hydrated grain.


Edited by Mateah (02/28/19 06:35 PM)


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Re: PC'd Grain - Life Span? [Re: Mateah] * 1
    #25844121 - 02/28/19 06:37 PM (2 months, 23 days ago)

Welll, when you say survive...

What I understand is that endospores can survive it.  They can be damaged yet repair themselves and produce bacteria but it takes a lot of time.


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Re: PC'd Grain - Life Span? [Re: BeefSupremeJr]
    #25844123 - 02/28/19 06:38 PM (2 months, 23 days ago)

Thanks Beef, for getting all the quotes together and saving me some time :smile: I'm not saying anything outlandish, just what becomes wicked evident when you search these forums and see what the TC's and legends like RR have to say. Learn from others mistakes and save yourself some time. I nocc'd up some two week old grain many years ago, and it all failed right quick. About five minutes in to "search function and chill" and I learned about why you want to noc grains immediately.


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25844127 - 02/28/19 06:40 PM (2 months, 23 days ago)

Quote:

Mateah said:
This is my story :typing:

Rye jar with SFD 120min 18 PSI about a month later and 4-5shakes later Nothing visible at all. This is a test jar.




I personally truly believe this rye in the jar is sterilized, but I leave room for other 'personal' experiences as well.




This does not indicate it's sterile or viable for our use. Endospores are designed to shell up to prepare for when the going gets tough and lie dormant until more favorable conditions arise. You're sure that no endospores survived and just haven't germinated because these conditions aren't favorable? There aren't endospores that might germinate if you inoculate with spore solution or spawn to a wet enough substrate or raise the temperature?

I'd love to see you inoculate and fruit these jars and compare to your standard results with a known culture. Or maybe you can sterilize a jar of water and dump it in the grain while it's still hot. I figure this would germinate any remaining (or collected after the cycle) endospores, right?


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Re: PC'd Grain - Life Span? [Re: sinisterminister7]
    #25844146 - 02/28/19 06:45 PM (2 months, 23 days ago)

Quote:

sinisterminister7 said:
Thanks Beef, for getting all the quotes together and saving me some time :smile: I'm not saying anything outlandish, just what becomes wicked evident when you search these forums and see what the TC's and legends like RR have to say. Learn from others mistakes and save yourself some time. I nocc'd up some two week old grain many years ago, and it all failed right quick. About five minutes in to "search function and chill" and I learned about why you want to noc grains immediately.





Thank mad season not me.  I just quoted him.

When I think of all the pain and years of error it took these legends to get where they are, I cringe.  Why would anyone want to go through what they already have? I’d rather pickup an easier hobby and quit this nonsense than endure all that. 

One of the finest horticulturalists I’ve ever worked for used to say “you and your fifteen fuckin  minutes of experience” to me— a veteran of 16 years.

And you better believe I shut my mouth and did what I was told. 
After literal years of just DOING what I was taught, I realized why.

You always go “Ooooooooooooh, so THATS why!”


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Edited by BeefSupremeJr (02/28/19 06:47 PM)


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Re: PC'd Grain - Life Span? [Re: BeefSupremeJr]
    #25844148 - 02/28/19 06:46 PM (2 months, 23 days ago)

Quote:

BeefSupremeJr said:
Welll, when you say survive...

What I understand is that endospores can survive it.  They can be damaged yet repair themselves and produce bacteria but it takes a lot of time.



Endosplres are not some terminator version of contaminants it's just a normal bacteria spore :shrug: many bacteria types produce so called endo spores but its still just a spore no different from another contaminant spore (as long as I'm aware)

And science tells us what these things can survive and what they can not live through. And they shouldnt be able to live through 115C+ Temps for more than 15-20 minutes. They simply can't survive this. Only if temps drop and raise again then they haven't been constant in that case I can see how they could be able to gain back the 'buffer' they find protection in when it comes to Temps and time.


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Re: PC'd Grain - Life Span? [Re: stareatclouds]
    #25844156 - 02/28/19 06:49 PM (2 months, 23 days ago)

Quote:

stareatclouds said:
Quote:

Mateah said:
This is my story :typing:

Rye jar with SFD 120min 18 PSI about a month later and 4-5shakes later Nothing visible at all. This is a test jar.




I personally truly believe this rye in the jar is sterilized, but I leave room for other 'personal' experiences as well.




This does not indicate it's sterile or viable for our use. I'd love to see you inoculate and fruit these jars and compare to your standard results with a known culture.



I Understand that it would be hard to verify that this rye is sterilized like how would one prove that, going through every micrometer of every grain inside out? Lol

So let me ask instead. What is hindering live cells and spores inside this rye to germinate and grow at this point? Is there a rational explanation for that in this case?


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Re: PC'd Grain - Life Span? [Re: BeefSupremeJr]
    #25844158 - 02/28/19 06:49 PM (2 months, 23 days ago)

Quote:

BeefSupremeJr said:
Quote:

sinisterminister7 said:
Thanks Beef, for getting all the quotes together and saving me some time :smile: I'm not saying anything outlandish, just what becomes wicked evident when you search these forums and see what the TC's and legends like RR have to say. Learn from others mistakes and save yourself some time. I nocc'd up some two week old grain many years ago, and it all failed right quick. About five minutes in to "search function and chill" and I learned about why you want to noc grains immediately.





Thank mad season not me.  I just quoted him.

When I think of all the pain and years of error it took these legends to get where they are, I cringe.  Why would anyone want to go through what they already have? I’d rather pickup an easier hobby and quit this nonsense than endure all that. 

One of the finest horticulturalists I’ve ever worked for used to say “you and your fifteen fuckin  minutes of experience” to me— a veteran of 16 years.

And you better believe I shut my mouth and did what I was told. 
After literal years of just DOING what I was taught, I realized why.

You always go “Ooooooooooooh, so THATS why!”



Thanks mad season :wink:


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25844160 - 02/28/19 06:50 PM (2 months, 23 days ago)

It’s all speculation mat.  Like I say, data speaks volumes.  I’d love it you were right, really I would. 
Rigor walks...


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Re: PC'd Grain - Life Span? [Re: BeefSupremeJr]
    #25844171 - 02/28/19 06:53 PM (2 months, 23 days ago)

I'm thinking about constructing an experiment that may put parts of this argument to rest. What if I was to make a really nice substrate with grain that doesn't risk of drying out but that is still shakable.

How many months would this fully hydrated nutritious substrate have to stand and look fresh before calling it sterile? Is there. A time frame for which some of you TCs would call such substrate sterilized?


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25844180 - 02/28/19 06:56 PM (2 months, 23 days ago)

Let’s say you hold a handful of jars for 50 days each. 

Make a poor boy syringe with them, and test it on 100 plates. 

That would satisfy me.


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Re: PC'd Grain - Life Span? [Re: BeefSupremeJr]
    #25844189 - 02/28/19 07:00 PM (2 months, 23 days ago)

I was thinking more like 90 days and then dropping a cube culture on it to see if it is able to fully colonize and fruit several flushes without a problem. That would satisfy me.


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25844194 - 02/28/19 07:02 PM (2 months, 23 days ago)

Quote:

Mateah said:
So let me ask instead. What is hindering live cells and spores inside this rye to germinate and grow at this point? Is there a rational explanation for that in this case?




Yes. There's a reason why your unprepped grain doesn't smell like old cheese fresh out of the bag, but if you soak a batch overnight and let it sit out, it stinks to high heaven. There are certain conditions that must be met to get \endocpores to germinate, just like anything else. If you left an empty petri dish open in your house, you wouldn't see molds and bacterial growth. But if you poured agar into it, you would. 

Bacterial endospores are not the same as regular spores. That's the entire point. I feel like you should actually do some reading on the subject.


Quote:

While significantly resistant to heat and radiation, endospores can be destroyed by burning or by autoclaving at a temperature exceeding the boiling point of water, 100 °C. Endospores are able to survive at 100 °C for hours, although the longer the number of hours the fewer that will survive.




And endospores in your grain require different sterilization methods than something like a scalpel in an autoclave at the hospital.


Quote:

Mateah said:
I'm thinking about constructing an experiment that may put parts of this argument to rest. What if I was to make a really nice substrate with grain that doesn't risk of drying out but that is still shakable.

How many months would this fully hydrated nutritious substrate have to stand and look fresh before calling it sterile? Is there. A time frame for which some of you TCs would call such substrate sterilized?




Your entire method of determining sterility is looking at the jar and not seeing green. Inoculating after 90 days and flushing 3 times wouldn't prove sterility, but would go a long way in proving the viability window is much, much wider than previously thought. Do that. It's what I suggested initially.


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25844196 - 02/28/19 07:03 PM (2 months, 23 days ago)

That’d do part of this, but still doesn’t prove that an endospore could be present.

Ok this is getting silly as fuck.


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Re: PC'd Grain - Life Span? [Re: stareatclouds]
    #25844219 - 02/28/19 07:13 PM (2 months, 23 days ago)

Quote:

stareatclouds said:
Quote:

Mateah said:
So let me ask instead. What is hindering live cells and spores inside this rye to germinate and grow at this point? Is there a rational explanation for that in this case?




Yes. There's a reason why your unprepped grain doesn't smell like old cheese fresh out of the bag, but if you soak a batch overnight and let it sit out, it stinks to high heaven. There are certain conditions that must be met to get \endocpores to germinate, just like anything else. If you left an empty petri dish open in your house, you wouldn't see molds and bacterial growth. But if you poured agar into it, you would.



Who said this rye wasn't prepped? It's hydrated just like the other jars that are colonizing. As I said it's a test jar.

So back to my original question, what would hinder live organisms and spores from germinating/growing in nutritious hydrated temperate environment?


Edited by Mateah (02/28/19 07:15 PM)


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25844234 - 02/28/19 07:20 PM (2 months, 23 days ago)

:facepalm:

I'm unwilling to get into another pointless back and forth with you where you ignore basic shit and misinterpret the majority of what I say. Especially when it doesn't seem like you actually know what endospores are or understand the concept of their survival and requirements for germination.

"Inoculating after 90 days and flushing 3 times wouldn't prove sterility, but would go a long way in proving the viability window is much, much wider than previously thought. Do that. It's what I suggested initially."

Do that. I'm not an expert on endospores, but I've suggested pouring sterile, hot water into the jar you posted, as well as inoculating and spawning very old jars to establish some sort of baseline for viability in your situation.


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Re: PC'd Grain - Life Span? [Re: stareatclouds]
    #25844257 - 02/28/19 07:30 PM (2 months, 23 days ago)

Maybe stop making it so personal every time? :confused:

I just want to discuss science and not what you think I should do with my life and time. And I'm looking for a productive discussion where we speak clear language.

Now I'm lead to believe that you think my rye jar is too dry to germinate contaminants? I've had this in the back of my mind too that's why I suggested the experiment with grain that is prepped and made I to a sub that doesn't dry out for 90 days, and keep temperated. What would in your opinion hinder live organisms/spores from growing/germinating in this substrate? Please just about the science and not about me. I do understand the difference of the composition of the outer shell of endospores and spores of molds. It's different more resistent but it still has its limits. That's why I said it's the same as other spores but I should have been clearer and said it's the same for our purposes we still go hard on the heat so we treat the endospores and mold spores the same when we sterilize grain. That's what I meant to say.

You are very firm on the notion that grain doesn't get sterilized but you can not answer the simple question of why wouldn't live organisms flourish in an optimal environment if the environment standards were met?


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25844308 - 02/28/19 07:55 PM (2 months, 23 days ago)

Nothing is personal, but you simply misunderstand far too much of what I say (and general mycology to my understanding) to make this worthwhile for me. In each response of yours, in this thread and others, there's 5+ things I disagree with that I'd have to quote specifically and address directly. All for you to reply back with 5 more things I disagree with and/or didn't say while ignoring initial facts.

I apologize if that's offensive or rude to you, but it is what it is. You were very clearly confused about the difference between endospores and regular spores and seem to be backtracking (poorly). No, you're not correct when you say endospores simply cannot survive more than 15-20 minutes at 115C. And you're obviously wrong by saying they're the same as every other spore.

I've suggested a decent enough test/experiment/whatever repeatedly. Good luck with whatever experiments you do.


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Re: PC'd Grain - Life Span? [Re: stareatclouds]
    #25844330 - 02/28/19 08:03 PM (2 months, 23 days ago)

*quietly backs away, and out the door*


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Re: PC'd Grain - Life Span? [Re: stareatclouds]
    #25844338 - 02/28/19 08:06 PM (2 months, 23 days ago)

Ive never been considered a thick or dense person by enyone, and I'm not disagreeing with anything you're saying I'm just asking you in your opinion what would hinder live organisms/spores from germinating/growing in optimal environment?

You have said I should pour sterilized water in my rye jar but you yourself know that that would negate the test immediately if a vector was presented :shrug:

And that's why I suggested the optimal sub that doesn't dry out, and I'm still asking you, what would hinder live cells from flourishing in this 'optimal' environment?

Everyone reading this thread can decide for themselves if I'm really that retarded that I'm considered pointless to talk to.. But I'm just wondering if you are thinking of a specific situation or not regarding my question.


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25844347 - 02/28/19 08:10 PM (2 months, 23 days ago)

:nosekiss:
Just kiss and make up already.


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Re: PC'd Grain - Life Span? [Re: Mr. Funguy]
    #25844362 - 02/28/19 08:18 PM (2 months, 23 days ago)

My theoretical and practical knowledge about home cultivation is perfectly reasonable by any standards on mush cult. I understand more or less the basics and have done a lot of reading. But there's always one person 'calling me out' for my poor understanding of this hobby and my poor results and technique. Always bringing old stuff up and making it extremely personal. This is not okay in my opinion, trying to undermine my posts by saying I'm obviously so thick I'm not worth talking to. This is so far from the truth that it's really sad reading it like that. I hope this changes in the future like I've said all the other times. I'll give it more time. And thanks for your inputs so far, maybe we're talking about different things or are confused about something else but if rather sort that out instead of resolving to creating further resistence to a productive discussion. This is my last 2 cents on this it's getting pretty tiresome... :crazy:


Edited by Mateah (02/28/19 08:19 PM)


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25844425 - 02/28/19 08:52 PM (2 months, 23 days ago)

Quote:

Mateah said:
My theoretical and practical knowledge about home cultivation is perfectly reasonable by any standards on mush cult. I understand more or less the basics and have done a lot of reading. But there's always one person 'calling me out' for my poor understanding of this hobby and my poor results and technique. Always bringing old stuff up and making it extremely personal. This is not okay in my opinion, trying to undermine my posts by saying I'm obviously so thick I'm not worth talking to. This is so far from the truth that it's really sad reading it like that. I hope this changes in the future like I've said all the other times. I'll give it more time. And thanks for your inputs so far, maybe we're talking about different things or are confused about something else but if rather sort that out instead of resolving to creating further resistence to a productive discussion. This is my last 2 cents on this it's getting pretty tiresome... :crazy:



Are you gonna do the experiment and let us all know how it turns out?


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Re: PC'd Grain - Life Span? [Re: sinisterminister7]
    #25844456 - 02/28/19 09:09 PM (2 months, 23 days ago)

Sure will as I always do! :thumbup:

I'm just trying to figure what criteria decides that a substrate is sterilized. That's why I'm interested in the opinions of TC's and other experienced growers.

I suggested that I make a grain based substrate that will stay hydrated and keep 'optimal' environment to living organisms and spores. If this substrate doesn't go 'bad' in any way even tho it's incubated in all sorts of 'contaminant promoting' environments such as higher Temps, humidity, water, lack of FAE and so on, can it not practically be considered sterile? If not then my follow up question is what requirements need to be filled for a grain substrate to be considered sterilized?

I'd like useful input on my idea of the experiment and I welcome critique cause I don't want to spend 3 months doing it for nothing.

What are the flaws of this idea for the experiment? I'm asking anyone at this point I'm just looking to widen my perspective on this experiment before doing it. I'm in it for the curiosity, I'll never stop being curious about this hobby.


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Re: PC'd Grain - Life Span? [Re: sinisterminister7]
    #25844459 - 02/28/19 09:10 PM (2 months, 23 days ago)

I've inoculated jars that have been pressurecooked 3 times over the course of a month and a half. I use MS and if after 2 weeks I don't see any results I repeat the whole process for that jar. Over time, I did notice some shrinkage, but I've never had a jar not colonize due to lack of moisture.


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Re: PC'd Grain - Life Span? [Re: Mr. Funguy]
    #25844473 - 02/28/19 09:18 PM (2 months, 23 days ago)

mateah i understand your assumption. That sterilized hydrated grains should exhibit signs of bacteria in a optimal enivronment. I disagree on what constitutes an optimal environment. Bacteria doesnt wanna grow on sterilized steam dried grains. Theres no excess water for them to flourish. The surface moisture dries off once you remove grains and shake them after a pc cycle. All the moisture is inside the grain. Mycelium can break that shit open and extract the moisture but bacteria is shit at it.

Visual confirmation is insufficient in determining the amount of bacteria present in “sterilized jars”. Bacteria is not as aggressive and more localized compared to cube mycelium. A sterilized properly hyrdated grain jar left unmolested, has a lower water content than a jar with ANY inoculant used. Agar, LI, and LC aRe sources of water. Introducing them will kick start the necessary conditions for bacterial growth that would otherwise lay dormant. Dont understimate the changes even a little water can do.

Most times i run into bacteria i see nothing visually. The tell tale sign of bacteria is time. Is your grain stalling? How many weeks? Did it stall after a shake? Thats bacteria.

Messing around with clean grains especially WBS from big name store where it has gone from factory to shelf  is pointless when looking to test the endospore theory. Millet is smaller and has less surface area to sterilize.

Heres my experince and my challenge to people not “believing” in endospores/ bacteria. Buy a 50# bag of oats or milo. Only buy a bag crawling with weevils. Dump the bag in a 5 gallon bucket and let the weevils multiply for a a few weeks. Normal pc times at 90 minutes become insifficient. It only buys you a 2 week grace period. Mold is not fucking the grains up. Or prep. Or sterile technique. 


Always bacteria showing up 4 weeks later and brining my jars to the slowest crawl across grains and they dont recover after a shake. The cleanest agar plates colonizing great for 2 weeks then....stall.

Someone thinking “oh all th endospores have been killed. It must be my prep or sterile technique” will be ruling out th grain source which it most certainly can be and is in certain situations.


First of all you need to set common parameters to be tested. Establish if grain is very dirty/some what dirty/ clean, soak time, inoculation method: liquid, solid, gelatin semisolid, how long before inoculation to allow endospores or bacteria to grow, expected  colonization % of myco quart, did the grain colonize quickly then stall?


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Re: PC'd Grain - Life Span? [Re: JHOVA]
    #25844491 - 02/28/19 09:25 PM (2 months, 23 days ago)

Well now I'm on a tight rope here. I would hate to waste a while dish on jars if it's not going to work... meh I guess I should just reprep them when the time comes. Damn that sucks.


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Re: PC'd Grain - Life Span? [Re: Vibe_Enthusiast]
    #25844495 - 02/28/19 09:27 PM (2 months, 23 days ago)

I would knock all the jars up with the cleanest agar you have. Might as well.


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Re: PC'd Grain - Life Span? [Re: Vibe_Enthusiast]
    #25844497 - 02/28/19 09:28 PM (2 months, 23 days ago)

And quite honestly. I do enjoy a nice debate and nicely worded responses to read.. as to adding to my knowledge.

So please.. continue lol


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Re: PC'd Grain - Life Span? [Re: JHOVA]
    #25844501 - 02/28/19 09:28 PM (2 months, 23 days ago)

That's why my suggestion is to use a grain based substrate instead of just grain. A mostly grain substrate that contains both field capacity substrate for molds, yiest and other shit and at the same time even contains excess water for bacteria to flourish. An all inclusive kind of sub is in my mind. Of course 'optimal' is 'optimal' hence the 'signs'. Im trying to eliminate variables like dry grain and substrates not supporting bacterial growth. That's why I'm fishing for opinions.

That's for chiming in J


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Re: PC'd Grain - Life Span? [Re: JHOVA]
    #25844504 - 02/28/19 09:29 PM (2 months, 23 days ago)

Quote:

JHOVA said:
I would knock all the jars up with the cleanest agar you have. Might as well.



Yeah man fuck it. I'll just keep the donor dish around because it grew out to be damn clean. But I'm waiting for the 2nd transfer to grow out.


Edited by Vibe_Enthusiast (02/28/19 09:29 PM)


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25844517 - 02/28/19 09:35 PM (2 months, 23 days ago)

Quote:

Mateah said:
That's why my suggestion is to use a grain based substrate instead of just grain. A mostly grain substrate that contains both field capacity substrate for molds, yiest and other shit and at the same time even contains excess water for bacteria to flourish. An all inclusive kind of sub is in my mind. Of course 'optimal' is 'optimal' hence the 'signs'. Im trying to eliminate variables like dry grain and substrates not supporting bacterial growth. That's why I'm fishing for opinions.

That's for chiming in J



The main point of substrate is that it's a digestible water retainer. If it's grain related I feel like that opens it up to contamination. Why not us poo, coir, or vermiculite?


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Re: PC'd Grain - Life Span? [Re: Vibe_Enthusiast]
    #25844520 - 02/28/19 09:37 PM (2 months, 23 days ago)

Thats introducing external variables. Keep it to grain and sterilized water, sterilized agar etc. if you add in a mixed sub what caused what? Yiu lose the plot.

The proper question frames the experiment.
Dirty grain is a thing to behold :rofl:


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Re: PC'd Grain - Life Span? [Re: JHOVA]
    #25844533 - 02/28/19 09:43 PM (2 months, 23 days ago)

Quote:

JHOVA said:
Thats introducing external variables. Keep it to grain and sterilized water, sterilized agar etc. if you add in a mixed sub what caused what? Yiu lose the plot.

The proper question frames the experiment.
Dirty grain is a thing to behold :rofl:



Are we still talking about whether or not grain dies over time in an unopened jar? By dying I mean unusable.


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Re: PC'd Grain - Life Span? [Re: JHOVA]
    #25844539 - 02/28/19 09:46 PM (2 months, 23 days ago)

Have you read what I've been discussing so far Mecha? :smile:

I'm trying to find out how one can determine that grain has been sterilized, or if it's even possible to determine this.

The grain based sub I'm thinking about contains both perfectly hydrated grain in one piece and also mushed grain and over hydrated grain, a nice mix of every kind of desired condition for live organisms and spores. :takingnotes:


Quote:

JHOVA said:
Thats introducing external variables. Keep it to grain and sterilized water, sterilized agar etc. if you add in a mixed sub what caused what? Yiu lose the plot.

The proper question frames the experiment.
Dirty grain is a thing to behold :rofl:



I'm not adding anything to anything post sterilization. I'm just thinking of ways of sterilizing (mostly) grain but in an environment that will not leave grains dry on the outside after 3 months. Like I said an all inclusive substrate with mostly grains. I prepare this sub in a qt jar and PC for 120min on 18 PSI and leave it for a few months to see if anything visible shows up. And maybe doing several of these subs one with pre soaked grain to promote germination/spore production pre sterilization cycle to see if that jar shows different results.

Feels like I'm being pretty straight forward with this idea, Idk what's causing all the confusion :lol:


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25844553 - 02/28/19 09:53 PM (2 months, 23 days ago)

Ugh... sterilize grain normally and let it sit out for 2 weeks, a month, 45 days, 60, etc. And inoculate with clean mycelium like you would any other jar. Who gives a fuck about proving whether it's sterile or not? Check and see if it's viable with a reliable culture. That's practical for our purposes. Fuck it, I have old grain sitting out now, I'll do it.


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25844558 - 02/28/19 09:55 PM (2 months, 23 days ago)

Quote:

Mateah said:
Have you read what I've been discussing so far Mecha? :smile:

I'm trying to find out how one can determine that grain has been sterilized, or if it's even possible to determine this.

The grain based sub I'm thinking about contains both perfectly hydrated grain in one piece and also mushed grain and over hydrated grain, a nice mix of every kind of desired condition for live organisms and spores. :takingnotes:


Quote:

JHOVA said:
Thats introducing external variables. Keep it to grain and sterilized water, sterilized agar etc. if you add in a mixed sub what caused what? Yiu lose the plot.

The proper question frames the experiment.
Dirty grain is a thing to behold :rofl:



I'm not adding anything to anything post sterilization. I'm just thinking of ways of sterilizing (mostly) grain but in an environment that will not leave grains dry on the outside after 3 months. Like I said an all inclusive substrate with mostly grains. I prepare this sub in a qt jar and PC for 120min on 18 PSI and leave it for a few months to see if anything visible shows up. And maybe doing several of these subs one with pre soaked grain to promote germination/spore production pre sterilization cycle to see if that jar shows different results.

Feels like I'm being pretty straight forward with this idea, Idk what's causing all the confusion :lol:



Just boil grain until 25% burst. You will get mush, over hydrated, and maxed in one go...
Then sterilize.
Or just sterilize a petri with agar the same way you would a jar after coughing on it a couple times. You will know soon enough is theres any germs or spores that survived.


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Re: PC'd Grain - Life Span? [Re: stareatclouds]
    #25844597 - 02/28/19 10:15 PM (2 months, 23 days ago)

Quote:

stareatclouds said:
Who gives a fuck about proving whether it's sterile or not?



A curious mushroom cultivator(s)? And it's not about "proving" anything to anyone or about making anyone wrong.


All I'm doing in this thread is trying to see if there exists a way to determine that a grain jar has been sterilized. And if there is a way to determine this then how would be the best way of approaching this problem of figuring this out. I have noted the idea of inoculating grains that have been stsrilized for a few weeks/months. I will probably inoculate my test rye jar as well.


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25844634 - 02/28/19 10:41 PM (2 months, 23 days ago)

anything you find isnt an indication of all grains. only your specific batch. to truly test any grain theory one must identify and experience dirty grain as well as clean as fuck grain.


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Re: PC'd Grain - Life Span? [Re: JHOVA]
    #25844674 - 02/28/19 11:07 PM (2 months, 23 days ago)

So.. my jars are just sitting on a shelf.. I have all sfd lids on each.

I'm just going to use these grains when the time comes and see what happens. I would think if the grains were 'sterile' I'll just have to hope the myc has enough moisture to grow?


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Re: PC'd Grain - Life Span? [Re: Vibe_Enthusiast]
    #25844677 - 02/28/19 11:09 PM (2 months, 23 days ago)

Trying my best to catch up on this reading. I'll read it all tomorrow lol.

I appreciate all of your replies nonetheless


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Re: PC'd Grain - Life Span? [Re: Vibe_Enthusiast]
    #25844776 - 02/28/19 11:56 PM (2 months, 23 days ago)

Quote:

Vibe_Enthusiast said:
So.. my jars are just sitting on a shelf.. I have all sfd lids on each.

I'm just going to use these grains when the time comes and see what happens. I would think if the grains were 'sterile' I'll just have to hope the myc has enough moisture to grow?



So are we just ignoring me?


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Re: PC'd Grain - Life Span? [Re: MechaLegend]
    #25844793 - 03/01/19 12:11 AM (2 months, 23 days ago)

If a jar is sterile it will remain so until opened or compromised in some way.  The easiest way to tell if a jar is sterile is to leave it out.  if nothing grows it's sterile.  alternatively you could spore test whatever vessel you are using for sterilization.  once the test is sent off an cultured the lab will report back as to whether it was in fact sterilized or not. Spore tests can be had mail order for relatively cheap.  Just make sure you follow the same SOP for the spore test that you would when running a load of grain.
  As for how long grain stays viable, that I cant comment on.  I dont see why they wouldnt remain viable for at least a couple weeks.  I'm with @stareatclouds, put a clean culture to it and see what happens.  A slice of agar is easier to replace than a sterile jar of grains.
I know I am new here an stuff but there seems to be some level of confusion surrounding sterilization with some folks here.  It seems some are apt to overthink things.  At the end of the day we aren't inventing the wheel, we are just trying to make our own using shit lying around our house and/or parts we get at dolla general.


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Re: PC'd Grain - Life Span? [Re: stareatclouds]
    #25844914 - 03/01/19 02:08 AM (2 months, 23 days ago)

Quote:

Mateah said:
Practically no spores and organisms we deal with can  survive 120C for 20 minutes.
The rest of the PC cycle time is spent on making sure this high temp has penetrated to the center of the jar. Let's say only 115C is able to reach into every grain. Still 15min of 115C kills the shit out of the contams we deal with usually. So I'm basically saying if the PC cycle is proper the grain should be sterilized. I make this statement mostly cause it's so unplausable to me to think of a scenario when a proper PC cycle wouldn't kill everything inside hydrated grain.




Exactly. There is no need whatsoever for 8+ or 24 hours to sterilize something (as was quoted earlier in the thread).


Quote:

stareatclouds said:
Ugh... sterilize grain normally and let it sit out for 2 weeks, a month, 45 days, 60, etc. And inoculate with clean mycelium like you would any other jar. Who gives a fuck about proving whether it's sterile or not? Check and see if it's viable with a reliable culture. That's practical for our purposes. Fuck it, I have old grain sitting out now, I'll do it.




I have done this many times without problems. I usually sterilize jars in bulk, and them let them sit until I need them.


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Re: PC'd Grain - Life Span? [Re: Pinback] * 1
    #25844927 - 03/01/19 02:27 AM (2 months, 23 days ago)

So can you share the specific prep, how long you waited, noc methods, and the results from each of the batches of jars?


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Re: PC'd Grain - Life Span? [Re: rm1024]
    #25844952 - 03/01/19 03:16 AM (2 months, 23 days ago)

Quote:

rm1024 said:
The easiest way to tell if a jar is sterile is to leave it out.  if nothing grows it's sterile.




This isn't true, though, unless we're not considering endospores alive because they're in stasis. If you jar up a quart of oats fresh from the bag, are you going to see visible growth? No. Is it sterile? No, it's just that the endospores haven't been introduced to the right conditions to influence activation and germination. But when you soak them in warm water, this catalyzes activation and germination (as well as more endospore formation).

It's possible that you don't kill EVERY endospore during the soak/simmer and PC cycle. If this is possible, it's also possible that your jar has endospores doing exactly what they're supposed to do: lie dormant in stasis until favorable conditions arise again and then activation and germination happens, i.e., higher moisture content than dried grains and warmer temperatures.

This is why I suggest dumping sterile water in a grain jar, but apparently we need absolutely 0 vectors. Okay then Mateah, sterilize a bag of grain with a jar of water inside it. After a while, unscrew the water through the bag and dump it over the grains and incubate. Let's see what happens.

Anyway, not all contaminations or bacterium will show visible growth after PCing as those aren't activation conditions. This is the entire crux of the issue. This is why I've repeatedly told Mateah that pointing at his jar like, "See, no weird growth or pin mold or anything!" is stupid. But again, Mateah is completely misunderstanding what endospores are and confusing them with regular mold spores which are easily killed off. No, Mateah, they ARE terminator spores. They're some of the most heat resistant organisms we know of.

If you can point me to anything that says a specific time frame and setting will kill 100% of endospores + whatever else, I'm all ears.

Here's some good information on optimal conditions to activate and germinate common endospores. And another here.


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Re: PC'd Grain - Life Span? [Re: stareatclouds]
    #25845064 - 03/01/19 06:26 AM (2 months, 23 days ago)

Quote:

stareatclouds said:
But again, Mateah is completely misunderstanding what endospores are and confusing them with regular mold spores which are easily killed off.



Really? You're saying that until today I didn't know that endospores were more resilient than other types of spores? Everyone in mush cult understands that I'm aware of why grain can't be steam sterilized in 90 min but BRF can, including you know this. So that means you're knowingly making false statements about me just to  to discredit me!?!? Way to go TC...


BTW I already explained why I expressed myself (if very Clumsy) about endospores being like other kinds of spores. Not that they are as easily killed as other spores, but that they also have their limitations like any other spore. And the setting in which we sterilize grain is aimed at killing endospores, the toughest ones in the jar. So we don't really consider "different types of spores" when sterilizing grain, for our purposes there is only one target group that needs to be focused on and the rest will die off along the way.


Now I'm of course totally open that some type of spores can survive for example 125C for 30 minutes. But that still doesn't mean that the grain can not be sterilized by turning up the heat to 135C and keeping for a lot longer. Do you see my point now I'm trying to make the whole time?

All organisms have their limitations when it comes to what they can endure without dying, if your sterilization setup meets the theoretical criteria for what is necessary to kill the organisms present, then why would you assume the grain hasnt been sterilized or even doubt it?

And if there is a limit for what the toughest spores can withstand, and you go far beyond that limit, then why would you not be fairly sure that you've killed the target group and sterilized the grain? (of course assuming you sterilized properly)


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Re: PC'd Grain - Life Span? [Re: Mateah] * 1
    #25845078 - 03/01/19 06:58 AM (2 months, 23 days ago)

Endospore survival can be enhanced by biofilms. In which layers of bacteria can protect themselves from sterilization in pockets. This is why scalpels etc.. need to be surgically clean before sterilization, otherwise it would take way longer than 15-20m

Our grains cant be cleaned at all really and any preexisting bacteria can make a biofilm and protect spores even further. To think a grain jar is 100% completely sterilized in 90-120m at 15psi is blasphemy to me. And should be to anyone whos done some research on endospores and sterilization. It would probably take over 8+ hours at pressure to ensure grain got sterilized in actuality. There's actually bacteria inside the grain that grew with it ever since the plant was an embryo, bacterial endosymbionts.

Some bacteria will form hydrophobic biofilms along with some proportion of the population making spores. The biofilm creates microscopic pores where water cant enter. Sort of like dipping a mushroom in water the water doesn't rush between the gills Because of the surface tension. So these little dry microscopic pockets of air protect the colony from steam penetration as well their insulating property alone helps protect.

Seems some people here have very strong opinions about endospores despite being brought up to speed about them


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25845093 - 03/01/19 07:15 AM (2 months, 23 days ago)

Quote:

Mateah said:
Really? You're saying that until today I didn't know that endospores were more resilient than other types of spores? Everyone in mush cult understands that I'm aware of why grain can't be steam sterilized in 90 min but BRF can, including you know this. So that means you're knowingly making false statements about me just to  to discredit me!?!? Way to go TC...


BTW I already explained why I expressed myself (if very Clumsy) about endospores being like other kinds of spores. Not that they are as easily killed as other spores, but that they also have their limitations like any other spore. And the setting in which we sterilize grain is aimed at killing endospores, the toughest ones in the jar. So we don't really consider "different types of spores" when sterilizing grain, for our purposes there is only one target group that needs to be focused on and the rest will die off along the way.


Now I'm of course totally open that some type of spores can survive for example 125C for 30 minutes. But that still doesn't mean that the grain can not be sterilized by turning up the heat to 135C and keeping for a lot longer. Do you see my point now I'm trying to make the whole time?

All organisms have their limitations when it comes to what they can endure without dying, if your sterilization setup meets the theoretical criteria for what is necessary to kill the organisms present, then why would you assume the grain hasnt been sterilized or even doubt it?

And if there is a limit for what the toughest spores can withstand, and you go far beyond that limit, then why would you not be fairly sure that you've killed the target group and sterilized the grain? (of course assuming you sterilized properly)




Ugh, I was never missing your point, Mateah. Your entire points are rooted in completely wrong information, such as the general behavior and purpose of an endospore. And you conveniently ignore relevant information to spout off more nonsense.

No, I don't think you understand endospores well when you think they'll all be killed by 20 minutes at 120c. And you clearly don't understand their ability to lie dormant and wait for better conditions when you consistently point to your dry grain jar as proof of sterility. Yeah, of course you think you're easily sterilizing your jars, dude. You're already at the wrong starting point in how it can be achieved and also think bacteria will always readily reveal itself.

First it's, "And they shouldnt be able to live through 115C+ Temps for more than 15-20 minutes. They simply can't survive this." Now it's, "Now I'm of course totally open that some type of spores can survive for example 125C for 30 minutes."

Okay, so your point is that at some point, if we reach the magic threshold that kills everything in the jar, everything will be killed and we'll reach true sterility? Yeah. No shit, dude. The entire point is that we don't know the exact numbers involved to truly sterilize. And a likely number of hours is not viable for us. Well, you seem to know that all endospores will be dead within 20 minutes of 115C+ temps. Please link a source on that.

I'm nowhere near an expert on endospores. There was a weekend I spent a chunk of time researching them, Tyndallization, reading bod's big thread on them, etc. And the number of things we still don't know about them yet is staggering. And tons of it is beyond my paygrade anyway. But I'm glad you figured it out.

Please "sterilize" a bag of grain for 20 minutes (or whatever) with a small jar of water in there. Once cool enough, unscrew it using the outside of the bag as handles and dump the hot water in the grains and let it sit. I'm curious how it'll smell in a day.


Edited by stareatclouds (03/01/19 07:22 AM)


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Re: PC'd Grain - Life Span? [Re: stareatclouds]
    #25845130 - 03/01/19 07:50 AM (2 months, 23 days ago)

Well this has turned into something:rolleyes:

My question was  mostly wondering if the grains were going to be okay in that time frame without drying out. I've clearly been awakened that bacteria is the main concern on top of the grains drying out. Me being new to this, and clearly very uninformed(until now).

Very different opinions here and I respect each and everyone of your debates and even going to the threshold and talking about trying it for yourselves. I PC'd the grain for 90 minutes @ 15 PSI(Rye berry grains).

I buy pretty top of the line Rye Berries, wash them thoroughly & PC (not saying that means much, but almost close to nothing comes off these grains when I wash them because they're already so "clean").

After reading its safe to assume that I either PC again before I inoc them.. but if that's the case I'm better off just taking the step and prepping more grain when the time is ready? I should have just waited and I pulled the trigger too fast because I had time on my hands to do so.

The jars are just sitting on my top shelf, SFD lids.. and weren't going to be handled until inoc. I was planning on putting plastic bags over the lids of them to suppress airflow so less evaporation happened for a few weeks til I was going to use them.

I feel stupid for wasting so much grain.. but I guess wasting grain + very clean agar wedges is more of a hit in the dick. That alone kind of makes me justify prepping all new grain a few weeks down the road.

I didnt consider bacteria actually being the problematic scenario here. I was more on it drying out.. but I'm glad that I have been informed that these jars may not be completely sterile and wasting a good agar wedge of bacteria is not worth my time nor effort.


Edited by Vibe_Enthusiast (03/01/19 07:51 AM)


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Re: PC'd Grain - Life Span? [Re: Vibe_Enthusiast]
    #25845137 - 03/01/19 07:54 AM (2 months, 23 days ago)

You should have tons of agar wedges ready to go. You can sterilize them and come back, rehydrate them if you want, and then go another 90 minutes, I bet. Just take this as an experiment and report back. I'll update with my results, as well.


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Re: PC'd Grain - Life Span? [Re: bodhisatta]
    #25845205 - 03/01/19 08:47 AM (2 months, 23 days ago)

Quote:

bodhisatta said:
Seems some people here have very strong opinions about endospores despite being brought up to speed about them



Idk if we're kind of stuck here so imma make one last effort to express my thoughts on this without confusing everyone with my out of style English :lol:

I understand that spores can be Inbedded in the center of a grain. Luckily grains are hydrated all the way and the heat usually reaches far. I also understand that some spores can form extra protective layers against harmful elements. But temperature and duration can also be increased to a point to where the protection doesn't work any longer. (theoretically)


So to me it seems like if you set out to sterilize your grain you will succeed almost every time. But if you set out to PC grain at 15-18PSI for 2h then you're not really even attempting to sterilize grain but rather to just kill off most organisms and cripple the rest for a little while... :shrug:

But then what is 'a little while'? :takingnotes:  I'm sry if people get irrtated by what they think/feel is irrelevant cencerns. That's highly subjective to what you put emphasis on. I personally am curious about what conditions make these crippled endospores come to life again. And would one determine that it was one of these germinating causing problems and not something else? I'm Not demanding answers I'm asking around to see what growers think about this.


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25845286 - 03/01/19 09:54 AM (2 months, 23 days ago)

I like to cook my jars in massive batches and sometimes it takes me 3-4 days to get around to knocking them all up. I've had some sit around 7+ days and majority of them still turned out fine.

So I know I can do it, and I don't lose too many jars, but there's no way to have a control to get any solid data about how long is too long. There's so many variables you'll never figure out what time period is "safe" to wait for inoculation after sterilization


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Re: PC'd Grain - Life Span? [Re: sh4d0ws]
    #25845313 - 03/01/19 10:19 AM (2 months, 23 days ago)

Quote:

stareatclouds said:
You should have tons of agar wedges ready to go. You can sterilize them and come back, rehydrate them if you want, and then go another 90 minutes, I bet. Just take this as an experiment and report back. I'll update with my results, as well.




Yeah man. I was working on it. Like I said, I'm a busy guy and I had some down time on Saturday to sit around the PC so I figured it would be nice to get that out of the way so when my agar is ready, I could drop &, use that jar for a g2g to all of the rest. I just got ahead of myself. Trying to be efficient kind of put a gamble on my hands that I'm not sure I want to take. Lol.
Quote:

sh4d0ws said:
I like to cook my jars in massive batches and sometimes it takes me 3-4 days to get around to knocking them all up. I've had some sit around 7+ days and majority of them still turned out fine.

So I know I can do it, and I don't lose too many jars, but there's no way to have a control to get any solid data about how long is too long. There's so many variables you'll never figure out what time period is "safe" to wait for inoculation after sterilization



Yeah. But 7 days and 4 weeks are two completely different sides of the spectrum if you ask me lol. I wouldn't have worried about a few days. Shit, its already been like 5 days since I've pc'd them. I guess I'll just see how it feels and go with my gut when the time comes. Like I said, I'm trying hard to get a cycle going.. with my agar to jars to shoebox ratio. I always want agar dishes and jars colonizing while my shoeboxes are fruiting.

Trying to get this rhythm going,and its starting up I just don't want to waste the first agar I ever use on grain that is not worthy lmao. But I do hate that I "wasted" this grain. I guess I could rehydrate and PC.. as long as it won't effect the grain doing that all over again.


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Re: PC'd Grain - Life Span? [Re: stareatclouds]
    #25845451 - 03/01/19 11:34 AM (2 months, 23 days ago)

Quote:

stareatclouds said:
So can you share the specific prep, how long you waited, noc methods, and the results from each of the batches of jars?




Sure. Typically, I weigh out 180 g of rye in a glass and add 180-210 g (depends on the rye) of water at 70-100 C. I put on a lid having a polyfill plug, and aluminium foil. Then, I pressure cook at 15 psi for 60-90 min, counting from when 15 psi is reached. After the pressure has gone down, I take out the jars and shake them while still hot, then allow them to cool in a clean place, which can be the pressure cooker.

I have also done it with pre-hydrated grains, with additives like gypsum, coffee waste and vegetable oil, and in filter bags, though most of my experience is like the paragraph above. Also bigger bags with sawdust, dung and straw, but then I have kept them longer at 15 psi (like 2-4 h) to allow for heat transfer. They last at least for several weeks.

The time I have waited with grain jars is at least several months (as written earlier in the thread). Right now, I have some filter bags with rye and coffee waste sitting that I sterilized February 12th. It is not that long, but they look great, and smell great through the filter.

My usual method of inoculation is with agar wedges.


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Re: PC'd Grain - Life Span? [Re: Pinback]
    #25845473 - 03/01/19 11:53 AM (2 months, 23 days ago)

Quote:

bodhisatta said:
To think a grain jar is 100% completely sterilized in 90-120m at 15psi is blasphemy to me. And should be to anyone whos done some research on endospores and sterilization. It would probably take over 8  hours at pressure to ensure grain got sterilized in actuality.




Why do you believe this?


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Re: PC'd Grain - Life Span? [Re: stareatclouds]
    #25845499 - 03/01/19 12:16 PM (2 months, 23 days ago)

Quote:

stareatclouds said:
Bacterial endospores are not the same as regular spores. That's the entire point. I feel like you should actually do some reading on the subject.




I found this thread of use, thanks for the discussion here between everyone.
Would you or anyone else be able to point me to any quality sources to do a bit of reading?


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Re: PC'd Grain - Life Span? [Re: Pinback]
    #25845528 - 03/01/19 12:31 PM (2 months, 23 days ago)

Ehhhh.. Fuck this experiment. Not even the internet knows how heat resistent Bacillus spores really are and I've read 3 PDFs so far on moist heat sterilization and Bacillus. 121C for 15min apparently kills the organism in vegetative state and destroys enough proteins and enzymes in the spore to render it more or less useless.

Bacteria obviously needs water to flourish and won't get very far on grain that is dry on the outside so right now I'm wondering, even if you have a healthy bacillus spore germinate in one of the grains what will become of it and can it spread to other grains in that environment?

Hmm..

After some back and forth I'm finally concluding that 2h on 18 PSI actually does accomplish what we're after, it makes the endospore sterile even if it doesn't kill it :shrug: (not in 100% of cases) but almost always :bongload:


Edited by Mateah (03/01/19 12:35 PM)


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25845558 - 03/01/19 12:46 PM (2 months, 23 days ago)

Not for ALL grains.


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Re: PC'd Grain - Life Span? [Re: JHOVA]
    #25845583 - 03/01/19 12:59 PM (2 months, 22 days ago)

The exception is the rule :eatingout: but still the exceptions are very rare so the rule kind of remains the rule lol.


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25845589 - 03/01/19 01:03 PM (2 months, 22 days ago)

Just had a jar of oats that was 11 days old contam.  Shook it and it was the brown mush. 
Smells like barbecue?

Second jar of oats on the same run to contam.  2hr pc cycle at 16psi. 

I find it doubtful that my lids are faulty.  Just saying.  I have a whole new set of lids and if this happens again the next run, I’m quitting oats.


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Re: PC'd Grain - Life Span? [Re: Pinback]
    #25845609 - 03/01/19 01:21 PM (2 months, 22 days ago)

Quote:

Pinback said:
Quote:

bodhisatta said:
To think a grain jar is 100% completely sterilized in 90-120m at 15psi is blasphemy to me. And should be to anyone whos done some research on endospores and sterilization. It would probably take over 8  hours at pressure to ensure grain got sterilized in actuality.




Why do you believe this?



See my other posts in this very thread


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Re: PC'd Grain - Life Span? [Re: bodhisatta]
    #25845744 - 03/01/19 02:24 PM (2 months, 22 days ago)

Thanks, but I don't see where you got 8 hours from. Microbiological waste (plates, solid media, cloth, tools, etc.) is routinely sterilized, in bags, in much shorter time.

If I would make petri dishes using semi-ground rye, and they didn't contaminate after two weeks, would that change your mind?


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25845752 - 03/01/19 02:30 PM (2 months, 22 days ago)

Absolute statements either apply universally or they arent absolute. Nothing wrong with nuance.


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Re: PC'd Grain - Life Span? [Re: JHOVA]
    #25845766 - 03/01/19 02:42 PM (2 months, 22 days ago)

You little bastards. :oldman:


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Re: PC'd Grain - Life Span? [Re: bodhisatta] * 2
    #25845889 - 03/01/19 03:40 PM (2 months, 22 days ago)

God damn it now I have even more questions. :shrug: These Bacillus are apparently mostly found in dirt/soil around the surface level and are present on most vegetation. 1g of human poop apparently contains thousands of these. There should theoretically be a bunch of them present in BRF as well since rice is vegetation and it comes from soil and is produced in open air. Bacillus spores are basically everywhere and must therefore end up in all sorts of organic material?

Quote:

bodhisatta said:
To think a grain jar is 100% completely sterilized in 90-120m at 15psi is blasphemy to me. And should be to anyone whos done some research on endospores and sterilization.



So what about my GWA plates? Would you consider them not sterile as well because they were PCd for only 15min at barely 5-10PSI on the ever degrading IKEA PC?

If I've understood you correct it's 'blasphemous' to believe that 100% of endospores are killed in 15min at 15PSI of 'effective heat cycle'. To me it sounds like you're inevitsbely implying also that a GWA media can not be sterilized, is this so and if not could you expand on this a little in case I've missed something essential? :takingnotes:

Id like your opinions and ideas as well Stare if you haven't been bored to death by now lol your posts are always well written and good explanations and good info relevant to the topic didcissed, that's why I always read all your posts I catch in MC. As you can see now I became curious about GWA and endospores, its just the way I am, want to know how is what and why. I don't agree with everything bod does or says or you or many other and God forbid if anyone should agree with everything I say and do. Most of us here are characters with strong personalities but I like how we can still keep it cool and have productive discussions without rendering our discussions pointless. I believe several posters in this thread have learned good stuff today. Peace

But yall please shed some light for me about GWA endospore sterilization? :popcorn:


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Re: PC'd Grain - Life Span? [Re: Mateah] * 2
    #25845917 - 03/01/19 03:51 PM (2 months, 22 days ago)

Liquids are extremely easily sterilized unlike grain. anything but liquids and surgically clean stainless steel instruments are a pain to sterilize. Grain water should take 15m @ 15psi.

A bioburden of 10,000,000 endospores in 500ml liquid would die to a survival chance of 1 in 10,000,000 with 15m @15psi. Now grain with a bioburden that's the same might take all day


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Re: PC'd Grain - Life Span? [Re: bodhisatta]
    #25845927 - 03/01/19 03:56 PM (2 months, 22 days ago)

It's not really a question of the endospores its a question of what material you're sterilizing. The endospores are there.

My theory about BRF is that the endospores are there they just cant do any harm so steaming works to kill all vegetative growth in cakes and that's sufficient


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Re: PC'd Grain - Life Span? [Re: bodhisatta]
    #25846000 - 03/01/19 04:34 PM (2 months, 22 days ago)

Quote:

bodhisatta said:
Liquids are extremely easily sterilized unlike grain. anything but liquids and surgically clean stainless steel instruments are a pain to sterilize. Grain water should take 15m @ 15psi.

A bioburden of 10,000,000 endospores in 500ml liquid would die to a survival chance of 1 in 10,000,000 with 15m @15psi. Now grain with a bioburden that's the same might take all day




If the endospores have a D121C of about 2 min, then yes. It is the same for grain, only that the slower heat transfer requires a longer time to reach 121 C.

As a general note, around 15 years ago, most people here sterilized grain for 1 hour, with good results. I don't know when that changed to recommend longer times.


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Re: PC'd Grain - Life Span? [Re: bodhisatta]
    #25846001 - 03/01/19 04:34 PM (2 months, 22 days ago)

Quote:

bodhisatta said:
Liquids are extremely easily sterilized unlike grain. anything but liquids and surgically clean stainless steel instruments are a pain to sterilize. Grain water should take 15m @ 15psi.

A bioburden of 10,000,000 endospores in 500ml liquid would die to a survival chance of 1 in 10,000,000 with 15m @15psi. Now grain with a bioburden that's the same might take all day



:takingnotes:

"Grain water should take 15min at 15PSI (~121C)" this part of the sterilization is what I call 'effective heat cycle' clocking from the point you've estimated that sufficient heat has reached out to all spores in the liquid media/grain jar.

As you mentioned it's much easier to heat an endospore in a liquid than one inside a wheat berrie that consists of ~75% water weight. Due to the lower density and pourus structure inside the grain, considerably higher temps than 121C are required to be able to heat the center of every grain to at least 121C. Or like you mentioned you would need much longer sterilization cycle to achieve the end goal of at least 15min of 121C of effective heat cycle.

But..

Here comes a honest question from me just trying to see if I'm on the right path of understanding all of this. So:


Let's say you are fairly sure that an 8h cycle on 15 PSI will deminish the bioburden inside grain jars down from 10 000 000 to 1. But what if you sterilized the grain at ~80 PSI, would those temps cut down the heat duration cycle to around 2-3h? You have an estimation off the hip for this?

Also I think I understand what you're saying about that I can't compare liquids and grain cause it's too different that simply increasing heat and duration 'linearly' won't guarantee every cubic mm in the grain will get cooked because of more complex variables in the composition of the grain/type of grain and prep. I could be wrong tho. :takingnotes: (I hope you can make sense of my last statement even tho I use concepts and dialectic in a little weird way cause I'm not totally rested.)


Edited by Mateah (03/01/19 04:39 PM)


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25846009 - 03/01/19 04:40 PM (2 months, 22 days ago)

Quote:

Mateah said:
Let's say you are fairly sure that an 8h cycle on 15 PSI will deminish the bioburden inside grain jars down from 10 000 to 1. But what if you sterilized the grain at ~80 PSI, would those temps cut down the heat duration cycle to around 2-3h? You have an estimation off the hip for this?





This can be modelled by a so-called z-value for the organism. But there really is no need for those temperatures and times...


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25846011 - 03/01/19 04:42 PM (2 months, 22 days ago)

Matea do you own a pc that goes to 80 psi? How about 300 psi? What about 10,000 psi?  The tools we have is a regular pc that can go up to 20 with a few quarters. Anything more is an arbitrary # pulled out the aether for practical purposes.


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Re: PC'd Grain - Life Span? [Re: JHOVA]
    #25846036 - 03/01/19 04:57 PM (2 months, 22 days ago)

@Pinback

Thanks for being European and that the link was in Celsius :hatsoff:
Fahrenheit has been giving me migraine lately, I can handle cups, ounces, feet and inches but my cpu is full for the moment when it comes to measurements in Hollywood language. :bongload:


Quote:

JHOVA said:
Matea do you own a pc that goes to 80 psi? How about 300 psi? What about 10,000 psi?  The tools we have is a regular pc that can go up to 20 with a few quarters. Anything more is an arbitrary # pulled out the aether for practical purposes.



My PC goes to 20 PSI, but autoclaves go higher and I might be able to get access to one or two soon. I could probably do something valuable by experimenting with sterilizing a bunch of jars at much higher Temps but not so high that they compromise the quality of the grain. But yes 80PSI was pulled out of gods beard for practical reasons :lol:


Edited by Mateah (03/01/19 04:58 PM)


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Re: PC'd Grain - Life Span? [Re: JHOVA]
    #25846052 - 03/01/19 05:09 PM (2 months, 22 days ago)

Quote:

JHOVA said:
Matea do you own a pc that goes to 80 psi? How about 300 psi? What about 10,000 psi?  The tools we have is a regular pc that can go up to 20 with a few quarters. Anything more is an arbitrary # pulled out the aether for practical purposes.



If you removed the safety blowout plug any PC would easily hold 80psi. I don't suggest it tho and you would burn the grains.


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Re: PC'd Grain - Life Span? [Re: bodhisatta] * 1
    #25846084 - 03/01/19 05:27 PM (2 months, 22 days ago)

planctonic(yes spelled right google will change it on you) micro-organisms are extremely easy to kill. Biofilms are a different ball game entirely. Planktonic means single cells disperse usually in liquid.

I do believe i once read that biofilms can make bacteria 1000x harder to sterilize than planctonic bacteria. Some of these biofilms contain endospores protected at the core. Bacteria in biofilms produces extracellular signaling chemicals that let subsets of the bacteria in the film do specific jobs like become more hydrophobic. Stranger yet is bacteria produce plasmids that float in the extracellular matrix in a biofilm these can enter other bacteria and change their DNA to perform specific tasks. So bacteria in a film behaves more like a multicellular organism than it resembles the single individual.

Poor pre-cleaning of medical instruments before sterilization is a big cause of failed sterilization. Think about trying to scrub the bioburden off of the inside of grains, you cant


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Re: PC'd Grain - Life Span? [Re: bodhisatta]
    #25846100 - 03/01/19 05:33 PM (2 months, 22 days ago)

:excuseme:


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Re: PC'd Grain - Life Span? [Re: bodhisatta]
    #25846115 - 03/01/19 05:41 PM (2 months, 22 days ago)

Quote:

bodhisatta said:
planctonic(yes spelled right google will change it on you) micro-organisms are extremely easy to kill. Biofilms are a different ball game entirely. Planktonic means single cells disperse usually in liquid.

I do believe i once read that biofilms can make bacteria 1000x harder to sterilize than planctonic bacteria. Some of these biofilms contain endospores protected at the core. Bacteria in biofilms produces extracellular signaling chemicals that let subsets of the bacteria in the film do specific jobs like become more hydrophobic. Stranger yet is bacteria produce plasmids that float in the extracellular matrix in a biofilm these can enter other bacteria and change their DNA to perform specific tasks. So bacteria in a film behaves more like a multicellular organism than it resembles the single individual.

Poor pre-cleaning of medical instruments before sterilization is a big cause of failed sterilization. Think about trying to scrub the bioburden off of the inside of grains, you cant





:whatwhatwhat:  :havesomescience:

Read today that Bacillus can die and still germinate if it's DNA is intact.


Edited by Mateah (03/01/19 05:41 PM)


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Re: PC'd Grain - Life Span? [Re: Mateah] * 2
    #25846118 - 03/01/19 05:42 PM (2 months, 22 days ago)

What's die really mean for a single celled bacteria when you can get it wet and it puts itself back together. Some of these signals that bacteria make during biofilm formation tell a subset of the film to protect their DNA and even further tell some bacteria to start forming extra tough endospores with more dipicolinic acid. As well some are forming with moderate levels and low levels. So that there's endospores that are super protected from DNA damage and some that are more ready to spring into action. Many people seem to think endospores only form in dire conditions. That just pushes the amount formed to a higher %. Bacteria is always maximize its survival chances. Some subset of the population of a film is making spores at a slow rate during optimal conditions and during dire conditions that proportion grows larger. This is why tyndalization doesn't really work and there's other reasons too like Never being able to germination every spore during the in between sterilization phases anyway.

I like to think of a biofilm in or on a grain as a fortified castle with a modern bank vault inside and inside the vault are people wearing the same gear that the people who work with molten steal do and if they die so long as not all their DNA is burnt to a crisp they can rebuild and come back to life.

So yea much different than sterilizing liquid


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Re: PC'd Grain - Life Span? [Re: bodhisatta]
    #25846153 - 03/01/19 06:02 PM (2 months, 22 days ago)

Quote:

bodhisatta said:

I like to think of a biofilm in or on a grain as a fortified castle with a modern bank vault inside and inside the vault are people wearing the same gear that the people who work with molten steal do and if they die so long as not all their DNA is burnt to a crisp they can rebuild and come back to life.

So yea much different than sterilizing liquid




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Re: PC'd Grain - Life Span? [Re: bodhisatta]
    #25846156 - 03/01/19 06:03 PM (2 months, 22 days ago)

Organismgasm

I knew from long ago they could monitor their environment and adapt while dormant, but I had no clue they were this komplex and interesting.

Aspergillus and Trich spores are like the Flintstones car and Bacillus is Transformers truck.


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25846167 - 03/01/19 06:10 PM (2 months, 22 days ago)

Bacteria gives no fucks


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Re: PC'd Grain - Life Span? [Re: JHOVA]
    #25870970 - 03/13/19 03:38 AM (2 months, 11 days ago)

So I got bored at page 2 of cultivation guru 1 says X and I reckon Y? And what would you know. . .

Can anyone fill me in on whether anyone used science here? Did we get to the bottom of this?

I PC'd 12 jars of rye 2 months ago.
PC'd for 2hr.
These have remained refrigerated with foil hats on.
Stored lying down
Food including dairy and meat Stored in same fridge.
Some grain water from the soak was left in the same fridge with holes in the lid and went green and fluffy (removed at 6 weeks, yep I pay heaps of attention)

I am now watching 3 x G2G jars colonise with p.allenii. . . Yet that ain't sciency enough right.

What say I inoculate some agar plates with individual grains from a few of the jars?

Wouldn't that (hides under chair) prove if anything was alive in there?


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Re: PC'd Grain - Life Span? [Re: Bobbit]
    #25870990 - 03/13/19 03:55 AM (2 months, 11 days ago)

You would in theory use the same grains soak water for agar. The reason I say same (same 50 lb bag) is because grain varies so fucking much.

Some things don't grow on lme and pda that grow on our grains.

That would probably be your best route imo


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Re: PC'd Grain - Life Span? [Re: van hatton]
    #25871052 - 03/13/19 04:41 AM (2 months, 11 days ago)

Well, I have some MEA poured, and not sure I'll soak any rye before making more agar as much of this thread tells me I'm wasting my time anyways...

However if there is a need to put speculation to sleep, then I may be convinced.


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Re: PC'd Grain - Life Span? [Re: Bobbit]
    #25871102 - 03/13/19 06:16 AM (2 months, 11 days ago)

No idea why you stored them in the fridge or what you think this proves either way? If you'd bother to read the thread, I think you'd have a better understanding of the issues and be able to answer your own questions.

Here are random thoughts:

  • There's a huge difference between "is this jar viable?" and "is every single living thing within this jar, including currently dormant endospores, rendered completely sterile?" And people keep interchanging them, making the debate a PITA.

  • The currently accepted idea of our "sterilization" is just killing enough vectors for a "window" where mycelium can get a head start. Thus, we believe that even WITH proper "sterilization" for mycology, we're still dealing with SOME bad guys (and obviously there are people debating that concept now).

  • If the above is correct, fruiting a jar successfully, no matter how long it sits after a PC cycle, doesn't prove everything in the jar was sterile. You don't need true sterility to spawn something and get fruits. People fruit bacterial jars all the time.

  • An endospore is specifically designed LIKE THE TERMINATOR and can withstand extreme conditions and lie in stasis until favorable germination conditions are met. Those conditions, to my knowledge, are primarily influenced by moisture (not dry), and temperature (not cold, not super hot).

  • If endospores survive your PC cycle, it's not likely they'll immediately germinate once it cools down to room temperature. As I said to Mateah, you don't open a bag of dry grains and puke from nasty bacteria stench. But if you soak your shit for 24 hours and leave it out a day or so, it reeks. Because those are conditions that catalyze the vegetative state (active bacteria), as well as endospore formation. So pointing to a dried out jar afterwards like, "See? Nothing!" is not indicative of everything being fine.

  • If the above is true, you'd be more likely to eyeball something's wrong when you introduce moisture and warmer conditions, like after inoculating. This is just my guess and could be wrong. And not all endospores are the same species and behave the exact same.

  • Dropping a single grain on agar doesn't speak for the entire jar. Why would it? Why can't another grain be harboring bacteria that wasn't killed like others? No idea if this is optimal, but I suggested repeatedly to mix in water to cover the grains and leave it within activation/germination temperatures. I think this would be a better test than a grain on agar. Google for endospore activation/germination conditions.

  • No matter how many times someone repeats any success with this, we aren't closer to knowing at which point in any given jar is truly sterile (or even if they WERE sterile). Original endospore count, grain type, prep method, sterilization time, PSI used, steam vented, and 3753 other things all make my situation different than yours. Maybe your grains are sterile and your inoculant isn't? Maybe my sterile technique sucks and yours is good? Perhaps I'm using shitty SFDs? Both things muddy our results, but aren't the fault of the grain. And again, even if it works, viability is different from sterility.

  • Personally, I care most about viability, and that's why I engage in these discussions. That's why I've just told people to try and use their old jars, not point at them. And while it's not concrete, I would feel better about using older grains if I knew that 20 other people had replicated this success over 100 jars each, at least establishing that the old idea of "a few days" was not as concrete with a certain baseline of time.

  • Related to above + people interchanging viability and sterility: while viability is most important, the science aspect of it is dope, and I wish I knew more. It would be amazing to know how many endospores were present AND the minimum time to truly sterilize. But we can't, and likely won't. And it's unnecessary for our purposes. So let's focus on those.

  • For what it's worth, I've used old grains successfully before. And they'd possibly been inoculated previously. I inoculated 5 WBS quarts with APE BLI. It showed no growth and sat on my shelf for 2 months (plastic lids IIRC). Curious if I even inoculated them in the first place, I smelled each of them and found no foul smell. So I resterilized them and inoculated them via AA+ G2G. They wound up sitting at 100% for 3-weeks on accident. I broke them up to spawn and somehow forgot them again so the grain starting recovering. I eventually spawned, cased, and fruited it on 5/26/18. The first flush was 26 days later, but average was 20 for that clone. Yield average for that clone/tub was 1407g. Our old, resterilized and neglected WBS was 1897g, a record for that variety for that spawn volume. No green or anything and smelled fine, but looked shot after harvest so I tossed it.

  • I've also used a bunch of older jars that looked fine with very poor results. No mold or obvious bacteria, but unreal low yields on proven cultures, which could be related to grain. But I'm not 100% either.

That was longer than I intended. And I could be wrong about any of it, so feel free to correct me if so! Also, anybody interested in documenting this? Then check this.


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Re: PC'd Grain - Life Span? [Re: stareatclouds]
    #25871342 - 03/13/19 10:57 AM (2 months, 11 days ago)

Another thought to consider is if grains could get truly sterile, why aren't there any grains freshly harvested, then canned? It would be nice af to get fresh, hydrated grains. It would then mean we wouldn't need to hydrate, and potentially if people did things right, wouldn't need to PC. However, it's clear that it won't work. Companies know grains won't last like that. They know that for long term storage, like in the span of years, drying them, or canning liquid form like oat meal is the way to do it.


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Re: PC'd Grain - Life Span? [Re: Mad Season]
    #25871374 - 03/13/19 11:32 AM (2 months, 11 days ago)

I had one jar leftover that I couldn't fit into the PC.. I just had to throw it out a few days ago because of green mold. The others seem and look fine.. also look hydrated still.

My agar will be finished by this weekend. I was going to drop my wedges to these grains... after all of this reading I'm not sure if that's a good idea or not lol. But they're already all prepped and do look fine. So I don't fucking know. Maybe I'm just better off prepping all new jars because I've read it could mean lower yields... though that's not completely backed up.

It will be about 15 days since PC'd. I don't think thats extremely long.. but two weeks vs 1 or 2 days...


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Re: PC'd Grain - Life Span? [Re: Mad Season] * 1
    #25871375 - 03/13/19 11:32 AM (2 months, 11 days ago)

They make canned corn. Canned oats or barley might not be a hot seller


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Re: PC'd Grain - Life Span? [Re: bodhisatta]
    #25871383 - 03/13/19 11:38 AM (2 months, 11 days ago)

True that. It's hard to say for sure, just a thought I was having the other day.


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Re: PC'd Grain - Life Span? [Re: Mad Season] * 1
    #25871463 - 03/13/19 12:22 PM (2 months, 11 days ago)

Canned grains are in liquid tho. Never loose hydrated grain like ours. No places to trap air if you've got it all in liquid


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Re: PC'd Grain - Life Span? *DELETED* [Re: stareatclouds]
    #25871592 - 03/13/19 01:40 PM (2 months, 11 days ago)

Post deleted by bodhisatta

Reason for deletion: If you're going to call people ignorant you can have your entire rebuttal deleted.


Edited by bodhisatta (03/13/19 02:44 PM)


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Re: PC'd Grain - Life Span? [Re: Bobbit]
    #25871669 - 03/13/19 02:27 PM (2 months, 10 days ago)

I spent quite a bit of time trying to recap relevant points to my understanding to catch you specifically up to speed. And you're being an ass. What are any of us going to have that is beyond circumstantial evidence?

We put plates and slants in fridges to slow down growth, yes, but that's for storing clean cultures. The point of these experiments is to figure out if you have bacteria/living things in your jars, not mask it by slowing life down. That's why it's recommended to let stuff grow out a bit before using from fridge, because in a few days, all the bacteria and pr mold spores and shit that wasn't showing will bloom.


Quote:

"what was I trying to prove?" Was more a verification process that being busy (read: absent minded and lazy) can still yield results, and at least some learning for myself, if not the community.




Great. I was asking for this exact clarification because people confuse sterility with viability and your goals would change my answer to you.

You're the only one being smug. You taking a single grain to an agar plate doesn't prove anything and isn't good science. How does taking 3 change anything? Did you skip over the previous discussion AND my bullet points?

If you're trying to test viability, why would sampling a few grains to agar do more than just using the jars? I suggested what I would do if not using them normally: dump sterile water in the grains and see what happens. Even if a single grain contaminated, that doesn't mean the whole jar would've, or even that the single grain would've had it been in the jar noc'd with mycelium and not your agar plate.


Quote:

I hope this encourages staresatignorance to get over their 'we'll never be able to tell so lets make it into a religion' stance.




Yeah, not like I'm the main dude campaigning for a large-scale documented test where we all record results with old grain.

The main point of my post is summed up in the first bullet point. And yet again, you completely miss it. go be lazy and absent minded when you're not involving others in it.


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Re: PC'd Grain - Life Span? [Re: stareatclouds]
    #25871694 - 03/13/19 02:40 PM (2 months, 10 days ago)

Quote:

stareatclouds said:
The currently accepted idea of our "sterilization" is just killing enough vectors for a "window" where mycelium can get a head start. Thus, we believe that even WITH proper "sterilization" for mycology, we're still dealing with SOME bad guys (and obviously there are people debating that concept now).



I have only ever seen that notion here. It is definitely not accepted in microbiology, food science, or pharmaceutical science. It is not a debate in those fields.


Quote:

If endospores survive your PC cycle, it's not likely they'll immediately germinate once it cools down to room temperature. As I said to Mateah, you don't open a bag of dry grains and puke from nasty bacteria stench. But if you soak your shit for 24 hours and leave it out a day or so, it reeks. Because those are conditions that catalyze the vegetative state (active bacteria), as well as endospore formation. So pointing to a dried out jar afterwards like, "See? Nothing!" is not indicative of everything being fine.



The jar can have enough moisture to support mycelial growth after several months. The drying (or non-drying) depends on relative humidity and mass transport of gas. And, if you kill vegetatively growing bacteria by heating to 100 C, you will see that endospores germinate quite quickly afterwards.


Quote:

Dropping a single grain on agar doesn't speak for the entire jar. Why would it? Why can't another grain be harboring bacteria that wasn't killed like others? No idea if this is optimal, but I suggested repeatedly to mix in water to cover the grains and leave it within activation/germination temperatures. I think this would be a better test than a grain on agar. Google for endospore activation/germination conditions.



So, you accept that individual grains can be sterilized? But not a bulk mass?


Quote:

No matter how many times someone repeats any success with this, we aren't closer to knowing at which point in any given jar is truly sterile (or even if they WERE sterile). Original endospore count, grain type, prep method, sterilization time, PSI used, steam vented, and 3753 other things all make my situation different than yours. Maybe your grains are sterile and your inoculant isn't? Maybe my sterile technique sucks and yours is good? Perhaps I'm using shitty SFDs? Both things muddy our results, but aren't the fault of the grain. And again, even if it works, viability is different from sterility.



Very good point, since apparently some people have contamination problems with (supposedly) sterilized jars. Clearly there are different opinions here. I believe most of the problems are on the operator side of the pressure cooker :smirk:


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Re: PC'd Grain - Life Span? [Re: Pinback]
    #25871718 - 03/13/19 02:49 PM (2 months, 10 days ago)

Just a reminder to everyone in general. if you need to flame your argument is probably shit and even if it isn't it's still getting dumped


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Re: PC'd Grain - Life Span? [Re: Pinback]
    #25871725 - 03/13/19 02:54 PM (2 months, 10 days ago)

Quote:

Pinback said:
Quote:

stareatclouds said:
The currently accepted idea of our "sterilization" is just killing enough vectors for a "window" where mycelium can get a head start. Thus, we believe that even WITH proper "sterilization" for mycology, we're still dealing with SOME bad guys (and obviously there are people debating that concept now).



I have only ever seen that notion here. It is definitely not accepted in microbiology, food science, or pharmaceutical science. It is not a debate in those fields.






It is definitely accepted in food science. Sterilization is a game of probabilities, only in food sciences you have the added constraint of maintaining taste.  This is why every relevant government agency suggests you limit how long you store canned goods for (e.g. https://www.cdc.gov/botulism/consumer.html), because no can is truly completely sterile.

And also in microbiology, you program your autoclave cycle against the most common expected contaminants. Every now and then sterilization fails 'cause you have something weird creep in, and you have to throw out your entire experiment, or you sequence something completely different.

For every time period, at temperature you experience a log reduction in the number of CFUs in the thing you are sterilizing.  Basic math tells you that you are unlikely to ever reach zero with a typical CFU load, but rather you take it down to an acceptable level without overly deteriorating the substrate.

It's also why you'll never ever find any sterilization method - beit filters or whatever - promising 100% sterilization rate. It's just not doable.

Something I've always wondered, but never tried or read: What actually happens when you sterilize grains for too long? Do they turn to literal mush?


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Re: PC'd Grain - Life Span? [Re: teladi]
    #25871728 - 03/13/19 02:55 PM (2 months, 10 days ago)

They dry out and go black


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Re: PC'd Grain - Life Span? [Re: Mad Season]
    #25871739 - 03/13/19 02:59 PM (2 months, 10 days ago)

Even though it's all steam?


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Re: PC'd Grain - Life Span? [Re: teladi] * 1
    #25871740 - 03/13/19 03:01 PM (2 months, 10 days ago)

:shrug: idk I've definitely left them in pc for way too long, and although it didnt run out of water, the grains were pretty much blackened af. Not like pure black, but pretty fuckin dark. Also yeah, they were much drier than they were going in.


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Re: PC'd Grain - Life Span? [Re: Mad Season]
    #25871776 - 03/13/19 03:23 PM (2 months, 10 days ago)

Quote:

Pinback said:
I have only ever seen that notion here. It is definitely not accepted in microbiology, food science, or pharmaceutical science. It is not a debate in those fields.




Do you have links to sterilizing grains with 100% certainty in those fields or any other?


Quote:

So, you accept that individual grains can be sterilized? But not a bulk mass?




I am assuming that not all parts of the jar/bag are always accessed equally by the forces needed to kill all endospores. And not all grains are created equally when it comes to endospore count. From my research, there's clear differences in thermal death times for different species of endospores. Is it not possible certain bits of grain have one and not the other? Is it not definite that grains on the outer wall of the bag get "sterilized" faster than those in the middle? So pulling one of those grains does not equal every single grain. And we don't know what the minimum time is to kill everything, so what's the point?

I have never said a bulk mass can't be sterilized, by the way.


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Re: PC'd Grain - Life Span? [Re: Mad Season]
    #25871785 - 03/13/19 03:28 PM (2 months, 10 days ago)

The way we prep our grain makes it (impossible?) to know wether everything inside has been killed or not because even if it survived it doesn't mean it will germinate and even if it germinates that doesn't mean that it will be able to multiply and spread into threatening numbers. So...

(imo)
To be able to produce any kind of valuable info from testing 'sterility' inside grain would be
to create some kind of grain test jar that has both nutrient rich water and grain inside of it.
That way we can be sure that spores that are still viable will be in 'optimal' environment to germinate
and multiply in.


Any thoughts on what's faulty about this kind of test jar? :takingnotes:


Edited by Mateah (03/13/19 03:29 PM)


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Re: PC'd Grain - Life Span? [Re: teladi]
    #25871806 - 03/13/19 03:48 PM (2 months, 10 days ago)

Quote:

teladi said:
It is definitely accepted in food science. Sterilization is a game of probabilities, only in food sciences you have the added constraint of maintaining taste.  This is why every relevant government agency suggests you limit how long you store canned goods for (e.g. https://www.cdc.gov/botulism/consumer.html), because no can is truly completely sterile.



That page is for home canning. "These outbreaks often occurred because home canners did not follow canning instructions, did not use pressure canners, ignored signs of food spoilage, or didn’t know they could get botulism from improperly preserving vegetables." Food dating is for food quality, not safety (see here )

Quote:

And also in microbiology, you program your autoclave cycle against the most common expected contaminants. Every now and then sterilization fails 'cause you have something weird creep in, and you have to throw out your entire experiment, or you sequence something completely different.

For every time period, at temperature you experience a log reduction in the number of CFUs in the thing you are sterilizing.  Basic math tells you that you are unlikely to ever reach zero with a typical CFU load, but rather you take it down to an acceptable level without overly deteriorating the substrate.

It's also why you'll never ever find any sterilization method - beit filters or whatever - promising 100% sterilization rate. It's just not doable.




No, sterilization at certain times will reduce the risk of contamination of a given item to some acceptable low number.

Please show me the basic math that says otherwise.


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Re: PC'd Grain - Life Span? [Re: Pinback]
    #25871913 - 03/13/19 04:45 PM (2 months, 10 days ago)

How is that sterilization?


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Re: PC'd Grain - Life Span? [Re: stareatclouds]
    #25871971 - 03/13/19 05:19 PM (2 months, 10 days ago)

I am not sure I understand your question. An acceptable number of contaminations due to incomplete sterilization for a hobby cultivator could be 1 jar per 100 (as an example). For food or pharmaceuticals, the acceptable contamination risk would be much lower.

Do we agree that, regardless of sterilization status, there is no short "inoculation window" due to microbial growth after heat treatment?


Edited by Pinback (03/13/19 05:33 PM)


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Re: PC'd Grain - Life Span? [Re: Pinback]
    #25872153 - 03/13/19 06:56 PM (2 months, 10 days ago)

Lol no I don't agree with that. If you had proof that said otherwise I would, but alas you've got 1 bag from february which apparently still smells fine. Big woop.

Here's a grain jar I pc'd for 50 mins that still smelled fine 2 months later AFTER inoculating it with a random bacteria on a plate.

It was shaken every 2 weeks.

Every person worth their salt who can actually identify bacteria based off of morphology of myc, and not the smell test which is useful like only half the time, has proper sterile tek with agar, yet seems to unarguably see more bacteria in spawn seems to agree that there's undoubtedly still bacteria germinating and thriving in their pc'd spawn. We all agree that the heat reached inside a PC should kill it. The thing which hasn't been proven, and bod said he wanted to try to prove is do the entire contents of the jars reach said temperature? Jars are filled with air pockets. Air, which is one of the greatest insulators.. I'd like to see an experiment done to actually put this to rest, but my experience, as well as countless other growers who've pushed thousands of spawn jars seem to unanimously agree something fishy is afoot.


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Re: PC'd Grain - Life Span? [Re: Mad Season]
    #25872430 - 03/13/19 08:55 PM (2 months, 10 days ago)

For instance gravity autoclaves will fail the bowie-dick test. Hence the Advent of vacuum pumped autoclaves


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Re: PC'd Grain - Life Span? [Re: bodhisatta]
    #25872450 - 03/13/19 09:05 PM (2 months, 10 days ago)

Can bacteria survive 90 minutes at 15 psi on grain? Yes
Does it always? No
Not all grain will be the same
That said grain is fucking cheap prep it when u need it


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Re: PC'd Grain - Life Span? [Re: Mad Season]
    #25872654 - 03/13/19 10:26 PM (2 months, 10 days ago)

:solved:


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Re: PC'd Grain - Life Span? [Re: Mad Season]
    #25873049 - 03/14/19 03:27 AM (2 months, 10 days ago)

Quote:

Mad Season said:
Lol no I don't agree with that. If you had proof that said otherwise I would, but alas you've got 1 bag from february which apparently still smells fine. Big woop.




No, I have done this many times over 15+ years. Sometimes inoculating directly after cooling, sometimes waiting weeks or months. Never a problem with bacterial growth. See an earlier post in this thread for my procedure.

What experiment (and outcome) would change your mind?


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Re: PC'd Grain - Life Span? [Re: Pinback]
    #25873151 - 03/14/19 07:00 AM (2 months, 10 days ago)

Something more compelling than anecdotes for starters

Soon ill test thermometers buried in grain. Air is heavier than steam so i really doubt our sterilization attempts and venting ever remove all the air.


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Re: PC'd Grain - Life Span? [Re: Pinback]
    #25873154 - 03/14/19 07:01 AM (2 months, 10 days ago)

Quote:

Pinback said:
What experiment (and outcome) would change your mind?



Idk who you were asking specifically but I'll chime in with my 2 cents..


After reading up some more on Bacillus propagation I can confirm that a liquid media is necessary indeed.
And needs to be incubated at 30-37C.


So.. Hydrated grain is NOT a liquid media and it is not kept anywhere near 37C.
So how is Bacillus propagation even supposed to occur in such test jars/environment? :shrug:



As I mentioned before it seems that my idea for test jars is what makes most sense to use in an experiment.
For example test jars can contain 500ml grain water + 300ml hydrated grain each.
And final test can be for example to use the Biopsy Method to inoculate
the test jars with a clean culture and wait for it to colonize the broth fully.


Does this make sense?


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25873156 - 03/14/19 07:04 AM (2 months, 10 days ago)

Additional water in a grain jar would make it far easier to sterilize. Especially if grain is submerged


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Re: PC'd Grain - Life Span? [Re: bodhisatta]
    #25873209 - 03/14/19 07:42 AM (2 months, 10 days ago)

I'm going to put these grains back into some water tomorrow and PC them again. That won't effect the grain in anyway, right? I just don't feel like wasting all thos grain I have in these jars.

I'm not going to chance wasting all these wedges on grain thats questionable.


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Re: PC'd Grain - Life Span? [Re: bodhisatta]
    #25873222 - 03/14/19 08:01 AM (2 months, 10 days ago)

Too bad we can just pour lcs on coir.

So I just woke up probably doesn't make much sense but what if we slightly hydrated grains during a short PC cycle 10-15 minutes drained access water then re pced.

The grains being submerged would make sterilization easier plus even with opening the jar to drain after that first run would probably be less shitty bacteria we can actually PC well.

Especially if inside of the grain the bacteria is dead. Each grain becomes it's own "container" which inside contains more sterile grains.

You following or am I stupid.

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Re: PC'd Grain - Life Span? [Re: bodhisatta]
    #25873233 - 03/14/19 08:15 AM (2 months, 10 days ago)

Quote:

bodhisatta said:
Additional water in a grain jar would make it far easier to sterilize. Especially if grain is submerged



GOOOOSHHHH!!!! I'll get back to this point later I've already thought about it..


But anyway.


So what are 'dirty grains' now again? To my understanding it is grains that contain a much higher endospore count.
These endospores>live bacteria require a liquid media to propagate in.
But hydrated grain is solid and NOT liquid, so how are these organisms supposed to propagate on grain and outcompete our live cultures? :takingnotes:


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Re: PC'd Grain - Life Span? [Re: bodhisatta]
    #25873473 - 03/14/19 11:28 AM (2 months, 10 days ago)

Sorry to interrupt this, ya'll are smarter than me on this, I'm just taking notes... however this quote struck me:

Quote:

bodhisatta said:
Additional water in a grain jar would make it far easier to sterilize. Especially if grain is submerged




Is there a reason we don't do this? I'm guessing we'll end up with oatmeal (if using oats) if we PC grains in water, even if we dump them right away after.

Interesting thought nonetheless.


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25873626 - 03/14/19 01:01 PM (2 months, 10 days ago)

Quote:

Mateah said:
Quote:

bodhisatta said:
Additional water in a grain jar would make it far easier to sterilize. Especially if grain is submerged



GOOOOSHHHH!!!! I'll get back to this point later I've already thought about it..


But anyway.


So what are 'dirty grains' now again? To my understanding it is grains that contain a much higher endospore count.
These endospores>live bacteria require a liquid media to propagate in.
But hydrated grain is solid and NOT liquid, so how are these organisms supposed to propagate on grain and outcompete our live cultures? :takingnotes:



The same way they do on agar:shrug:


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Re: PC'd Grain - Life Span? [Re: cronicr]
    #25873644 - 03/14/19 01:10 PM (2 months, 10 days ago)

My finding - I have experimented with this for quite sometime. I tried to use grain jars past the 48he mark that everyone goes by. I have had some luck with it. I find that if u are opening the jar after 2-3 day mark it normally molds BUT if u are NOT opening the lid to knock them up like using a syringe thru a self healing I section port  with either LI or LC then it seem to work ok. :thumbup:


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Re: PC'd Grain - Life Span? [Re: Apc123]
    #25873750 - 03/14/19 01:53 PM (2 months, 9 days ago)

So say I had cooked up a batch of rye jars last weekend but didn't get around to inoculating all of them right away, and put them in the fridge (sealed jars with plastic lids with SFDs, still foiled) the same day. If I were going to knock up more jars this weekend---so 1 week later---would it be better to (a) use them as-is or (b) PC them again, even if that risks over-cooking or throwing off water content?

(And yes, obviously fresh grain would be best, but I'm open to a slightly higher failure risk in exchange for not wasting the grain. So just want to know which option you all think would be more likely to succeed. FWIW, these will be going into shoeboxes / other small containers, so one bad jar won't ruin the whole grow - hence why I'm ok with a bit of risk. Thanks!)


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Re: PC'd Grain - Life Span? [Re: Durgin]
    #25873780 - 03/14/19 02:05 PM (2 months, 9 days ago)

Quote:

Mateah said:
Quote:

bodhisatta said:
Additional water in a grain jar would make it far easier to sterilize. Especially if grain is submerged



GOOOOSHHHH!!!! I'll get back to this point later I've already thought about it..


But anyway.


So what are 'dirty grains' now again? To my understanding it is grains that contain a much higher endospore count.
These endospores>live bacteria require a liquid media to propagate in.
But hydrated grain is solid and NOT liquid, so how are these organisms supposed to propagate on grain and outcompete our live cultures? :takingnotes:



By this theory we would be fine to pasturize grain
Quote:

Durgin said:
So say I had cooked up a batch of rye jars last weekend but didn't get around to inoculating all of them right away, and put them in the fridge (sealed jars with plastic lids with SFDs, still foiled) the same day. If I were going to knock up more jars this weekend---so 1 week later---would it be better to (a) use them as-is or (b) PC them again, even if that risks over-cooking or throwing off water content?

(And yes, obviously fresh grain would be best, but I'm open to a slightly higher failure risk in exchange for not wasting the grain. So just want to know which option you all think would be more likely to succeed. FWIW, these will be going into shoeboxes / other small containers, so one bad jar won't ruin the whole grow - hence why I'm ok with a bit of risk. Thanks!)



For a whole week i wouldnt even use the fridge


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Re: PC'd Grain - Life Span? [Re: cronicr]
    #25873810 - 03/14/19 02:18 PM (2 months, 9 days ago)

Quote:

cronicr said:
For a whole week i wouldnt even use the fridge




Thanks for reassurance. I thought that was probably the case, but figured the fridge wouldn't hurt anything except maybe moisture, and unlike contamination you can at least see if moisture is off right away.


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Re: PC'd Grain - Life Span? [Re: Pinback]
    #25873939 - 03/14/19 03:42 PM (2 months, 9 days ago)

Quote:

Pinback said:
Food dating is for food quality, not safety (see here )





I don't refer to food dating of best before dates, but expiration dates found on many, many canned goods. Expiration dates are a thing, but don't seem to be a thing in your country.

Quote:

Pinback said:

No, sterilization at certain times will reduce the risk of contamination of a given item to some acceptable low number.

Please show me the basic math that says otherwise.




I don't think you are understand what I am saying. Please re-read the sentence:

Quote:

It's also why you'll never ever find any sterilization method promising 100% sterilization rate. It's just not doable.




But you seem to be contradicting yourself. You first say, "I have only ever seen that notion here" referring to stareatclouds saying you can't sterilize completely, and then say, "will reduce the risk of contamination of a given item to some acceptable low number.". Not sure if you realize this.

Eh, missed an entire conversation in between, sorry guys for throwing the flow out.


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Re: PC'd Grain - Life Span? [Re: teladi]
    #25873941 - 03/14/19 03:44 PM (2 months, 9 days ago)

Quote:

teladi said:
Quote:

Pinback said:
Food dating is for food quality, not safety (see here )





I don't refer to food dating of best before dates, but expiration dates found on many, many canned goods. Expiration dates are a thing, but don't seem to be a thing in your country.

Quote:

Pinback said:

No, sterilization at certain times will reduce the risk of contamination of a given item to some acceptable low number.

Please show me the basic math that says otherwise.




I don't think you are understand what I am saying. Please re-read the sentence:

Quote:

It's also why you'll never ever find any sterilization method promising 100% sterilization rate. It's just not doable.




But you seem to be contradicting yourself. You first say, "I have only ever seen that notion here" referring to stareatclouds saying you can't sterilize completely, and then say, "will reduce the risk of contamination of a given item to some acceptable low number.". Not sure if you realize this.

Eh, missed an entire conversation in between, sorry guys for throwing the flow out.



Expiration dates and best by dates are guidelines put out by the company. They really have nothing to do with actual expiration.


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Re: PC'd Grain - Life Span? [Re: bodhisatta]
    #25873952 - 03/14/19 03:54 PM (2 months, 9 days ago)

Best by, for sure, but expiration dates are less than guidelines and have very specific definitions, in many regions. It's still dictated by the manufacturer, and based on when the manufacturer expects the goods to be unsafe for consumption.


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Re: PC'd Grain - Life Span? [Re: teladi]
    #25873968 - 03/14/19 04:12 PM (2 months, 9 days ago)

Most expiration dates are more for the packaging then the food or contents


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Re: PC'd Grain - Life Span? [Re: teladi]
    #25873982 - 03/14/19 04:19 PM (2 months, 9 days ago)

Quote:

teladi said:
Best by, for sure, but expiration dates are less than guidelines and have very specific definitions, in many regions. It's still dictated by the manufacturer, and based on when the manufacturer expects the goods to be unsafe for consumption.



Not really. Like cron said usually its packaging material or quality of food. Not safety.


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Re: PC'd Grain - Life Span? [Re: ShaperDreaming]
    #25874039 - 03/14/19 04:46 PM (2 months, 9 days ago)

[disclaimer]Just wanna say I really enjoy piecing together this puzzle together with you fellow mushroom people.
I hope my multitude of questions and statements don't cause irritation or nothing I'm just inquiring about this subject because it's interesting.[/disclaimer] :eatingout:


Alright, lets get inflammatory then..  :doggull:


It feels like I'm getting (or interpreting), somewhat inconsistent theories on the process of heat sterilizing endospores. I'm just trying to separate what we KNOW from what we 'have faith in' or 'most likely is so'.


Consensus/authorities around here hint to us that endospores are not just resilient as fuck, but you'd almost be silly to assume that 15min on 15PSI (effective heat cycle) will kill them completely. But then again I don't see anyone stressing about GWA or GWLC that's been sterilized at 15PSI for 15min :shrug: This makes me wonder why are these endospore containing nutrient rich LIQUIDS considered sterilized in 15min but the same endospores in grain (that sustain the same 'effective heat cycle') are only considered 'sufficiently incapacitated'. I have previously asked about if one can not simply just increase the pressure(heat) and the duration of the cycle to be able to effectively sterilize grain, and I was left with the impression that endospores are quite simply too tricky and too unpredictable that we'll ever be able to say that a grain jar has been effectively sterilized. And yet grain is FAR from optimal for propagating these organisms while nutrient rich liquid media's are pretty CLOSE to optimal environments for these organisms.



I get it that a grain jar possibly contains much higher spore count than GWA or GWLC and thus increases the chances of spores surviving. But this then begs the question; are there any statistics on these 'REALLY really thug life' endospores that somehow survive prolonged high temps while 99.99% of their buddies die with their mouths open already even before 15 min? (I understand that conditions can be environmental as well, like dry pockets between/inside grain) but I also know that there is no reason one shouldn't be able to always reach 'desired tepms/duration' within each grain if properly prepared and properly executed sterilization cycle.

(meaning: if your goal is to reach effective heat cycle of 15min/121C when sterilizing liquid media, then all you need to do is boil the liquid media under pressure for 15min. 15PSI makes the boiling point of water will rise to 121C. So you have permiated to every cell with 'desired temps'. But if you're sterilizing hydrated solids like grain inside qt jars then you're also dealing with partial insulation. From what I can compute so far the only practical solution would be to raise the temperature and prolong the heat cycle to compensate for the insulation. Let's say 15PSI allows for 114C to reach into the center of every grain within an hour. So you raise the pressure for example to 20-30PSI and add 30min more to the cycle and you're guaranteed that more than 114C will permeate every grain.


(I don't mean to sound so sure or cocky, I'm neither (I hope) everyone should read  my statements as mere observations/opinions/speculations, I'm just grateful for other takes on it and for clearing up facts for me if I happened to misunderstand something :takingnotes:


Okej here's another question I can't find the answer to online, is Bacillus subtilis fairly homogenous across individual organisms when it comes to heat resistence or can the differences vary greatly? Obviously weaker than average spores are no concern, but how stronger are the 'stronger than average' if they even exist? Yeah this is my question for now :bongload:


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25874051 - 03/14/19 04:53 PM (2 months, 9 days ago)

Like ive said everywhere and numerous times liquids are extremely easy to sterilize. Gravity autoclaves have a hard time getting air out they fail the bowie-dick test unlike vacuum pumped autoclaves.

They make spore indicators. They're loaded with 10^7 or more endospores. They get put in the middle of a load in the middle of an autoclave. 15m at 15psi kills them to a sterility assurance level of 10^-7 chance of a single survival. But we are sterilizing grain which is completely different.


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Re: PC'd Grain - Life Span? [Re: bodhisatta]
    #25874149 - 03/14/19 05:36 PM (2 months, 9 days ago)

Thanks for the response and so you know you're not wasting your time either lol, I have a private journal notebook where I save useful posts like your last post here, Ive read numerous of your posts about the strips since way back, very appreciative of your patience!

I understand there are different types of Bacillus organisms and different types of grain. I'm mostly asking around if anyone knows about specific numbers and shit  when it comes to the science of killing and incapacitating endospores with heat sterilization?


I've been looking online for quite a bit now, downloaded some PDF's and read also about endospore sterilization and propagation on a forum this week but I can't seem to find this specific information anywhere :shrug:


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Re: PC'd Grain - Life Span? [Re: teladi]
    #25874266 - 03/14/19 06:38 PM (2 months, 9 days ago)

Quote:

teladi said:
But you seem to be contradicting yourself. You first say, "I have only ever seen that notion here" referring to stareatclouds saying you can't sterilize completely, and then say, "will reduce the risk of contamination of a given item to some acceptable low number.". Not sure if you realize this.





Apologies if I was not clear, let me try again. I have only ever seen here the notion that an item cannot be completely sterilized (or that it would take more than 8 or 24 hours at 121 C) to do so. Yes, there are bacteria that will survive long times at 121 C, but they are typically found in hydrothermal vents and are not relevant here.

For practical purposes, an item is sterilized for a time that is calculated or estimated to reduce the risk of contamination to an acceptable level. That level will vary depending on what is sterilized.

Suppose, as an example, that you have a 1 log reduction of some bacterium every minute at 121 C, and that there are 100 viable bacteria in every jar you want to sterilize. The bacterial count is unrealistic, but will show my point.

After 1 minute at 121 C, there are 10 viable bacteria, per jar, on average (1 log reduction).
After 2 minutes, there is 1 viable bacterium.
After 3 minutes, there is 1 bacterium in every 10th jar.
After 4 minutes, there is 1 bacterium in every 100th jar.
After 5 minutes, there is 1 bacterium in every 1000th jar. And so on. This is what the model statistically predicts, and will naturally differ in reality. For example, heating times and heat transfer has to be taken into account.

So, sterilizing 1000 jars for 5 minutes according to the example, 999 of them are sterile (again, statistically and according to the model) and 1 of them has 1 viable bacterium. This is not contradicting what I wrote earlier.


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Re: PC'd Grain - Life Span? [Re: Pinback]
    #25874799 - 03/14/19 11:48 PM (2 months, 9 days ago)

Quote:

Pinback said:
Quote:

And also in microbiology, you program your autoclave cycle against the most common expected contaminants. Every now and then sterilization fails 'cause you have something weird creep in, and you have to throw out your entire experiment, or you sequence something completely different.

For every time period, at temperature you experience a log reduction in the number of CFUs in the thing you are sterilizing.  Basic math tells you that you are unlikely to ever reach zero with a typical CFU load, but rather you take it down to an acceptable level without overly deteriorating the substrate.

It's also why you'll never ever find any sterilization method - beit filters or whatever - promising 100% sterilization rate. It's just not doable.




No, sterilization at certain times will reduce the risk of contamination of a given item to some acceptable low number.

Please show me the basic math that says otherwise.




This is why I said, "How is that sterilization?"

Sterilization in microbiology is killing ALL forms of life, including currently active, as well as dormant endospores. Something is either considered sterile or not. Nobody would say 95% sterile. My syringes and needles I aspirate LC into come sterile. You're saying they still have bacteria and/or mold spores on them, just at a low enough amount to be called "sterile?" Or are you just splitting hairs that we'll never truly know 100%?

Why would a filter announce a sterilization % in the first place? They don't sterilize anything, they prevent things from passing through.

I don't really understand what your argument is and seems to flip-flop back in forth, but maybe I'm just tired. I DO 2 HOURS 16-20PSI almost every time. I don't know the starting bioburden, but this works for me. :smile:


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25874804 - 03/14/19 11:51 PM (2 months, 9 days ago)

Quote:

Mateah said:
I understand there are different types of Bacillus organisms and different types of grain. I'm mostly asking around if anyone knows about specific numbers and shit  when it comes to the science of killing and incapacitating endospores with heat sterilization?


I've been looking online for quite a bit now, downloaded some PDF's and read also about endospore sterilization and propagation on a forum this week but I can't seem to find this specific information anywhere :shrug:




Whoa! If only a handful of people had repeatedly said exactly this repeatedly throughout the thread? Come on Mateah, if modern science can't document this conclusively, your coir-water-grain-vessel contraption is needed now more than ever.


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Re: PC'd Grain - Life Span? [Re: Pinback]
    #25874857 - 03/15/19 12:19 AM (2 months, 9 days ago)

Quote:

Pinback said:
Sure. Typically, I weigh out 180 g of rye in a glass and add 180-210 g (depends on the rye) of water at 70-100 C. I put on a lid having a polyfill plug, and aluminium foil. Then, I pressure cook at 15 psi for 60-90 min, counting from when 15 psi is reached. After the pressure has gone down, I take out the jars and shake them while still hot, then allow them to cool in a clean place, which can be the pressure cooker.

I have also done it with pre-hydrated grains, with additives like gypsum, coffee waste and vegetable oil, and in filter bags, though most of my experience is like the paragraph above. Also bigger bags with sawdust, dung and straw, but then I have kept them longer at 15 psi (like 2-4 h) to allow for heat transfer. They last at least for several weeks.

The time I have waited with grain jars is at least several months (as written earlier in the thread). Right now, I have some filter bags with rye and coffee waste sitting that I sterilized February 12th. It is not that long, but they look great, and smell great through the filter.

My usual method of inoculation is with agar wedges.




How are your rye bags?


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Re: PC'd Grain - Life Span? [Re: stareatclouds]
    #25874920 - 03/15/19 01:01 AM (2 months, 9 days ago)

Maybe you think my test jar 'contraption' is ridiculous but I bet that I can get still get away with attaining at least some useful data from experimenting with sterilizing liquids, grain, and other contraptions at different temperatures and durations to later inoculate the test jars with sterile water or LC or agar cultures, depending on which hypothesis I'm trying to confirm or deny..


For example bod mentioned repeatedly that liquids sterilize completely different from grain, I get that.
Of course overhydrated grains submerged in water will conduct heat more like a liquid and less like insulated dry solid.



That's why my idea for another 'contraption' test jar is to sterilize grain in one jar and GW in another one and then add the sterilized GW to the grain jar. That way I won't make the sterilization of the grain more effective but I will still give the surviving endospores on that grain a chance to colonize a nutritious liquid broth if it wants to. See? I worked around that problem cause I'm constantly thinking about this experiment and I'm motivated by curiosity.



I'm way past noob and I have been for a while, but that doest mean that I won't be asking the same question again maybe in two years If I have a two year break from growing. Life throws curve balls you know :bongload: I'm not trying to prove anyone wrong or prove anything about anyone. I can also get frustrated when my long time JJ students ask me to show them basic techniques that I already showed them a bunch of times, but then again it's not my place to question their personal dedication to the art, I'm there just as a friend/guide showing them how it's done (how I do it). But if students ask me to prepare them for a tournament then I'll be more of a general and less of a friendly guide. But out of competition It's always very relaxed setting :bongload:


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25874954 - 03/15/19 01:18 AM (2 months, 9 days ago)

posting to read through later


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Everything i say is completely hypothetical...


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Re: PC'd Grain - Life Span? [Re: stareatclouds]
    #25875071 - 03/15/19 03:09 AM (2 months, 9 days ago)

Quote:

stareatclouds said:
Sterilization in microbiology is killing ALL forms of life, including currently active, as well as dormant endospores. Something is either considered sterile or not. Nobody would say 95% sterile. My syringes and needles I aspirate LC into come sterile. You're saying they still have bacteria and/or mold spores on them, just at a low enough amount to be called "sterile?" Or are you just splitting hairs that we'll never truly know 100%?




No, I completely agree that what we call sterile has no viable organisms. My point is that, for practical purposes, there will always be a small but acceptable risk of something being non-sterile after a sterilization procedure. Not that a sterile item itself has some small but acceptable number of bacteria. Do you see the difference?

My rye bags were subsequently inoculated, with no sign of contamination as far as I can tell.


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25880450 - 03/17/19 06:01 PM (2 months, 6 days ago)

Quote:

Mateah said:
Maybe you think my test jar 'contraption' is ridiculous but I bet that I can get still get away with attaining at least some useful data from experimenting with sterilizing liquids, grain, and other contraptions at different temperatures and durations to later inoculate the test jars with sterile water or LC or agar cultures, depending on which hypothesis I'm trying to confirm or deny..


For example bod mentioned repeatedly that liquids sterilize completely different from grain, I get that.
Of course overhydrated grains submerged in water will conduct heat more like a liquid and less like insulated dry solid.



That's why my idea for another 'contraption' test jar is to sterilize grain in one jar and GW in another one and then add the sterilized GW to the grain jar. That way I won't make the sterilization of the grain more effective but I will still give the surviving endospores on that grain a chance to colonize a nutritious liquid broth if it wants to. See? I worked around that problem cause I'm constantly thinking about this experiment and I'm motivated by curiosity.



I'm way past noob and I have been for a while, but that doest mean that I won't be asking the same question again maybe in two years If I have a two year break from growing. Life throws curve balls you know :bongload: I'm not trying to prove anyone wrong or prove anything about anyone. I can also get frustrated when my long time JJ students ask me to show them basic techniques that I already showed them a bunch of times, but then again it's not my place to question their personal dedication to the art, I'm there just as a friend/guide showing them how it's done (how I do it). But if students ask me to prepare them for a tournament then I'll be more of a general and less of a friendly guide. But out of competition It's always very relaxed setting :bongload:



Have you looked into any papers from doc holiday or aloha?
They have done by far the most research and these days do a hydrate and light sterilizing then 24 hours latwr do 4 hours at 60 psi....yes 60


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Re: PC'd Grain - Life Span? [Re: cronicr]
    #25880491 - 03/17/19 06:25 PM (2 months, 6 days ago)

:takingnotes: cheers!

Edit: OK I tried searching 3 different doc holiday's, nothing and found no Aloha either.. But both sound familiar tho..


Edited by Mateah (03/17/19 07:09 PM)


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Re: PC'd Grain - Life Span? [Re: Mateah]
    #25881257 - 03/18/19 03:17 AM (2 months, 6 days ago)

You can read about him here, here or in comments here :smirk:


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Re: PC'd Grain - Life Span? [Re: Pinback]
    #25881352 - 03/18/19 06:27 AM (2 months, 6 days ago)

Quote:

Pinback said:
No, I completely agree that what we call sterile has no viable organisms. My point is that, for practical purposes, there will always be a small but acceptable risk of something being non-sterile after a sterilization procedure. Not that a sterile item itself has some small but acceptable number of bacteria. Do you see the difference?

My rye bags were subsequently inoculated, with no sign of contamination as far as I can tell.




Right, and the acceptable risk is that window we have to introduce mycelium to overtake the substrate in case anything is leftover. Either way, everyone's situation is different. You should document these month old grows, for sure. Check the link in my signature.


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Re: PC'd Grain - Life Span? [Re: stareatclouds]
    #25881366 - 03/18/19 06:55 AM (2 months, 6 days ago)

John Holliday not doc Holliday. His doctorate is honorary. I wouldn't call him a doctor.

Aloha does some weird stuff like double sterilization.


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Re: PC'd Grain - Life Span? [Re: bodhisatta] * 1
    #25890889 - 03/22/19 09:19 PM (2 months, 1 day ago)

"listen you fuckers. you screwheads. here is a man who would not take it anymore. a man who stood up against the scum, the cunts, the dogs, the filth, the shit. here is someone who stood up." - travis bickle

who's in charge here? has this community project yet begun? also where do i send cash? i'm almost at my foto upload limit.

intent: an n=1 experiment to investgate the envelope of prepared grain jar shelf life.

six jars total. racehorse oats. bod prep. jars cooked no later than 2/1, probably the week before. can't recall the exact date.

sterilizer cycle: automated control, varying between 17 and 22 psi for two hours following a ten minute vent. allowed to fully cool before opening the sterilizer and checking lids for stiction.

stored, uncovered and unrefrigerated, on a rack in the lab since removing from the sterilizer.

inoculated 3/12 with a half plate each of a T2 culture from a wild cube print. tar shaken all the way out of them.

2/1-3/12 is about 5.5 weeks. fotos taken 3/22. ten days post inoculation.





anticipate mid colonization shake in three to five days.

just decided to move this to a journal. i get annoyed having to cruise long threads to piece together particular projects.


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Re: PC'd Grain - Life Span? [Re: J. Jack Flash]
    #25890932 - 03/22/19 10:02 PM (2 months, 1 day ago)

:creepylurker:


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Re: PC'd Grain - Life Span? [Re: ttching8475]
    #25903432 - 03/29/19 09:19 AM (1 month, 26 days ago)

20190327
all but one jar seem just fine. a couple are probably as good as i've ever done. that one just didn't colonize very much. still some, and it looked fine, just not quick.
shook them all tonight.



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Re: PC'd Grain - Life Span? [Re: J. Jack Flash]
    #25903467 - 03/29/19 09:49 AM (1 month, 26 days ago)

Thank you for your service, sir.


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Re: PC'd Grain - Life Span? [Re: Durgin]
    #25903483 - 03/29/19 10:04 AM (1 month, 26 days ago)

balls to that. ;] skeptics are going to say it means nothing, no matter what the outcome.

:highfive:

just trying to keep this thought alive. people been talking about doing this. stareatclouds is talking about formalizing this kind of experimentation. decided instead of asking how/if this is going, i'm doing my own thing, and hopefully others do the same. or at least the leaderishtypepeople see there's interest and consider lending their expertise and constructive criticism. meanwhile, i'm bascially threadjacking.

:classictroll:


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OfflineDurgin
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Re: PC'd Grain - Life Span? [Re: J. Jack Flash]
    #25903503 - 03/29/19 10:34 AM (1 month, 26 days ago)

I'm eager to see it play out. I knocked up about 15 jars a couple weeks ago that had been sitting in the fridge for 2-3 weeks, and so far I've lost one to mold but the rest are going strong. That seems like a relatively normal result (and if anything, I'd blame my noob skills for the one failure), but time will tell. Most of them are being used for sclerotia, so they're going to be hanging out for a while, and if the waiting hurt them it'll come out eventually.


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InvisibleJ. Jack Flash
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Re: PC'd Grain - Life Span? [Re: Durgin] * 2
    #25904840 - 03/29/19 10:48 PM (1 month, 25 days ago)

good on ya, man. all in this together.

here's about 48 hours after the shake.



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OfflineDurgin
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Re: PC'd Grain - Life Span? [Re: J. Jack Flash]
    #25905030 - 03/30/19 01:29 AM (1 month, 25 days ago)

Someday I hope you'll share how you get your grains to look so same damned perfect. They're like the Abercrombie models of grains.


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InvisibleJ. Jack Flash
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Re: PC'd Grain - Life Span? [Re: Durgin]
    #25905411 - 03/30/19 09:58 AM (1 month, 25 days ago)

easy. straight bodhisatta oats method. no rinse. dried overnight-ish on a giant pet screen elevated a few inches above a giant table. 4.5lbs dry oats for ten 1-liter mason jars. would use less for g2g receiving jars, but i don't do g2g. who needs that much mush?


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InvisibleJ. Jack Flash
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Re: PC'd Grain - Life Span? [Re: J. Jack Flash]
    #25908384 - 03/31/19 08:45 PM (1 month, 23 days ago)



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Re: PC'd Grain - Life Span? [Re: J. Jack Flash]
    #25908728 - 04/01/19 01:02 AM (1 month, 23 days ago)

Awesome recovery!


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InvisibleJ. Jack Flash
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Re: PC'd Grain - Life Span? [Re: kratom_redmomd]
    #25909069 - 04/01/19 09:04 AM (1 month, 23 days ago)

yeah it's looking pretty ok. culture A is faster than the other two, and one of these jars was noticably behind the rest when i shook them. since i'm make-pretending to be a scientist about this, and obviously don't really know what i'm doing, i decided to shake them and spawn them all together, rather than at some subjectively determined %colonization. :shrug: they're 19 days since inoculation in the last foto.


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InvisibleJ. Jack Flash
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Re: PC'd Grain - Life Span? [Re: J. Jack Flash]
    #25912432 - 04/03/19 12:13 AM (1 month, 21 days ago)



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OfflineDurgin
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Re: PC'd Grain - Life Span? [Re: J. Jack Flash]
    #25912443 - 04/03/19 12:17 AM (1 month, 21 days ago)

Looking nice and clean. Love the super bright white myc.


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Re: PC'd Grain - Life Span? [Re: J. Jack Flash]
    #25912960 - 04/03/19 09:03 AM (1 month, 21 days ago)

What variety is that?


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InvisibleJ. Jack Flash
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Re: PC'd Grain - Life Span? [Re: Vibe_Enthusiast]
    #25913005 - 04/03/19 09:29 AM (1 month, 21 days ago)

it's from a wild cubensis print sent to me from a member. i'm told it was collected on a family ranch somewhere in south carolina.

i'm trying to stifle any hope or emotional engagement in this project. i'm not a scientist, but i play one on the internet.


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InvisibleJ. Jack Flash
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Re: PC'd Grain - Life Span? [Re: J. Jack Flash]
    #25918484 - 04/05/19 11:17 PM (1 month, 18 days ago)

3/29


3/31


4/2


4/5



spawning soon. planning to put each jar in its own shoebox at about 1:2 straight coir, bodhi prep. probably in the tent to fruit.


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Re: PC'd Grain - Life Span? [Re: J. Jack Flash]
    #25918640 - 04/06/19 01:30 AM (1 month, 18 days ago)

That really is straight-up grain porn.


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InvisibleJ. Jack Flash
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Re: PC'd Grain - Life Span? [Re: Durgin]
    #25919256 - 04/06/19 01:03 PM (1 month, 18 days ago)

hey thanks man. they do look better than any i've done before. i guess it's safe to conclude that the age of those jars didn't impede colonization in any meaningful way. even the straggler is near ready to spawn.


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Re: PC'd Grain - Life Span? [Re: Mad Season]
    #25919523 - 04/06/19 04:20 PM (1 month, 17 days ago)

Quote:

Mad Season said:
Another thought to consider is if grains could get truly sterile, why aren't there any grains freshly harvested, then canned? It would be nice af to get fresh, hydrated grains. It would then mean we wouldn't need to hydrate, and potentially if people did things right, wouldn't need to PC. However, it's clear that it won't work. Companies know grains won't last like that.




how much would you be willing to pay? probably not enough to cover the costs of the lengthy processing time they would need compared to the usual tins of fully submerged food, large tins of food need significantly longer times. There is not much of a market, and as you know some companies do sell PF jars or bags of hydrated grain, for high prices. I had a thread asking about treating dry grains, they would be cheaper to ship as they are not hydrated or in heavy tins. I wondered if they could be irradiated dry, and then a simply hydration and steaming might be enough for us to use. I also wondered if roasted grains might be feasible to just steam and use, I have grown on highly roasted grains before. I was recommending "easy cook" dry rice before as it has already usually gone through a heat treatment so I reckon it would have a better chance.

We already can easily get suitable hydrated grains which are not fully submerged -pouches of cooked rice. I have seen numerous threads of grows with them with some success. I have colonised tinned peas, corn and kidney beans before, I emptied them out and rinsed and microwaved them in jars to both heat them as they were exposed to contams -but also to drive off moisture. Hippie3 grew on tinned hominy. I see other grains in pouches these days, but they would be too expensive for anybody doing a big grow.

I have done quite a few tests of double heat treating, like cooking whole grains fully submerged in water at 15-28psi, when while still hot taking them out and loading jars and reheating them, either steaming but usually microwaving. If you weigh your grains you can calculate what water content you are at. By driving off moisture after the initial heat treating it means they can have higher hydration to begin with so I think it should help.

Some grains are more suitable, like rye grass seed, which is very thin and does not really explode when overhydrated and/or exposed to high pressure. You can cook up a load of it in a PC, then strain it off and load into jars. Moisture can be squeezed out but I find it still stays too hydrated. These could be steamed or microwaved. If you need to drive off moisture they could be placed on heating pads. This is far too much hassle for many people but might suit some people, like myself. It could be used as a test jar as discussed, leave it highly hydrated for a long time and see what happens, then dry it out months later on a heat pad or oven. I have found fully sealed jars which must be over a decade old which look OK.

somebody asked what happens to grain cooked for a long time, or high psi.

this is popcorn in a sealed jar with no filter


after 19hours sitting in water at 97C


and after 115hours at 97C


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InvisibleJ. Jack Flash
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