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OfflineVibe_Enthusiast
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Re: PC'd Grain - Life Span? [Re: Vibe_Enthusiast]
    #25844677 - 02/28/19 09:09 PM (5 years, 30 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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OfflineMechaLegend
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Re: PC'd Grain - Life Span? [Re: Vibe_Enthusiast]
    #25844776 - 02/28/19 09:56 PM (5 years, 30 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 - 02/28/19 10:11 PM (5 years, 30 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 12:08 AM (5 years, 30 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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Invisiblestareatclouds
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Re: PC'd Grain - Life Span? [Re: Pinback] * 1
    #25844927 - 03/01/19 12:27 AM (5 years, 30 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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Invisiblestareatclouds
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Re: PC'd Grain - Life Span? [Re: rm1024]
    #25844952 - 03/01/19 01:16 AM (5 years, 29 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 04:26 AM (5 years, 29 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: Mateja] * 1
    #25845078 - 03/01/19 04:58 AM (5 years, 29 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: Mateja]
    #25845093 - 03/01/19 05:15 AM (5 years, 29 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 05:22 AM)

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OfflineVibe_Enthusiast
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Re: PC'd Grain - Life Span? [Re: stareatclouds]
    #25845130 - 03/01/19 05:50 AM (5 years, 29 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.


--------------------
:greyalien:



Edited by Vibe_Enthusiast (03/01/19 05:51 AM)

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Invisiblestareatclouds
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Re: PC'd Grain - Life Span? [Re: Vibe_Enthusiast]
    #25845137 - 03/01/19 05:54 AM (5 years, 29 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 06:47 AM (5 years, 29 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: Mateja]
    #25845286 - 03/01/19 07:54 AM (5 years, 29 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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OfflineVibe_Enthusiast
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Re: PC'd Grain - Life Span? [Re: sh4d0ws]
    #25845313 - 03/01/19 08:19 AM (5 years, 29 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 09:34 AM (5 years, 29 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 09:53 AM (5 years, 29 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 10:16 AM (5 years, 29 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 10:31 AM (5 years, 29 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 Mateja (03/01/19 10:35 AM)

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Re: PC'd Grain - Life Span? [Re: Mateja]
    #25845558 - 03/01/19 10:46 AM (5 years, 29 days ago)

Not for ALL grains.


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Re: PC'd Grain - Life Span? [Re: JHOVA]
    #25845583 - 03/01/19 10:59 AM (5 years, 29 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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