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Invisibletrauma47645
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Sterilzation of grain by soaking?
    #5497022 - 04/09/06 03:51 PM (17 years, 9 months ago)

Would it be possible to soak grain type substrates (WBS, rye berries) in a bleach or H2O2 solution to kill all the endospores that would otherwise survive PC'ing? Maybe doing a soak then rinsing the sterilization agent off the subtrate. Then putting it in a PC for the called amount of time. (Maybe this is a more reliable way than fractional sterilization to make it possible to steam sterilize grains for those that dont have PC's)


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Invisiblepong
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Re: Sterilzation of grain by soaking? [Re: trauma47645]
    #5497029 - 04/09/06 03:53 PM (17 years, 9 months ago)

if h2o2 worked i will be extremely happy.

has anyone tried it?


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Invisibletrauma47645
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Re: Sterilzation of grain by soaking? [Re: pong]
    #5497039 - 04/09/06 03:55 PM (17 years, 9 months ago)

I was hoping someone would be able to tell me if it would work


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Offlineskeletor
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Re: Sterilzation of grain by soaking? [Re: trauma47645]
    #5497118 - 04/09/06 04:30 PM (17 years, 9 months ago)

i really do not think it will but you could try a small batch and make sure and kill the myth. yaya im currently watching mythbusters ahah


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Offlinemogur
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Re: Sterilzation of grain by soaking? [Re: skeletor]
    #5497233 - 04/09/06 05:22 PM (17 years, 9 months ago)

The reason H2O2 won't work is that complex carbs and enzymes inactivate it. The only way that I know to make grains compatible with peroxide is to break down the enzymes in grain by heat sterilization. If you just throw peroxide in with untreated grains, you will get a bunch of fizzing and no peroxide left. In theory there should be an amount of H2O2 that would break down the enzymes and have just enough left over to attack the contams and thereby sterilize the grain, but there's no known practical way to determine that precise amount, and any left over would attack the mushroom spores. Mycelium is self-protecting from peroxide, because it will produce enzymes that breakdown the peroxide in the vicinity of the mycelium, but spores don't have that protective mechanism available.

I don't know about bleach, but I assume it would not rinse off, and even if it did, the rinse would have to be sterile, so what is gained? Also, both H2O2 and bleach are not all that effective against endospores, so other problems aside, heat is both more effective and easier.


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Invisibletrauma47645
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Re: Sterilzation of grain by soaking? [Re: mogur]
    #5497265 - 04/09/06 05:35 PM (17 years, 9 months ago)

bleach isnt effective against endo spores? what about microwaving the grain.. wouldnt microwave radiation kill them or ateast render them unable to reproduce?


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Offlinemogur
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Re: Sterilzation of grain by soaking? [Re: trauma47645]
    #5497312 - 04/09/06 05:56 PM (17 years, 9 months ago)

Quote:

From this link:
Hypochlorite has been used as a disinfectant for more than 100 years. It has many of the properties of an ideal disinfectant, including a broad antimicrobial activity, rapid bactericidal action, reasonable persistence in treated potable water, ease of use, solubility in water, relative stability, relative nontoxicity at use concentrations, no poisonous residuals, no color, no staining, and low cost. The active species is undissociated hypochlorous acid (HOCl). Hypochlorites are lethal to most microbes, although viruses and vegetative bacteria are more susceptible than endospore-forming bacteria, fungi, and protozoa.




Microwaving is a viable option, and blackout is the guru in that area. It's not the microwaves themselves that are destructive to contams, however, since once again, the effective agent is the heat produced by the microwaves.


Edited by mogur (04/09/06 06:00 PM)


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Invisibletrauma47645
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Re: Sterilzation of grain by soaking? [Re: mogur]
    #5497323 - 04/09/06 05:58 PM (17 years, 9 months ago)

so it would still kill endospores... im just trying to develope a non PC method for sterilizing grain for those who dont have one. (Even though I do have a PC)


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InvisibleFooMan
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Re: Sterilzation of grain by soaking? [Re: trauma47645]
    #5497347 - 04/09/06 06:11 PM (17 years, 9 months ago)

Quote:

trauma47645 said:
so it would still kill endospores... im just trying to develope a non PC method for sterilizing grain for those who dont have one. (Even though I do have a PC)




You can sterilize whole grains without a PC if you grind them down like BRF in the PF tek. If you can't PC, go PF method. If you are trying to grow alot of mushrooms, spawn the cakes to straw, compost or manure.

Like mogur said, even if you did find some sort of solution to soak the grains in that actually killed all contaminants including endospores, you'd still have to find a sterile way to rinse the same solution off of the grains before inoculating.

I'm not trying to knock the idea though. It would be great to not have to PC grains  :grin:


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Offlinemogur
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Re: Sterilzation of grain by soaking? [Re: trauma47645]
    #5497356 - 04/09/06 06:16 PM (17 years, 9 months ago)

Microwaving and fractional sterilization are definite possibilities, as many have reported positive results. Blackout has had many successes with Microwaves and I have been playing around with fractional sterilization with encouraging, but mixed results so far. I think if someone put enough effort into objective experiments, that a reliable method could be developed. Less likely are methods that involve simple treatment with disinfecting solutions, but then, who knows for sure without trying it.


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Invisiblemusher_420
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Re: Sterilzation of grain by soaking? [Re: mogur]
    #5497368 - 04/09/06 06:21 PM (17 years, 9 months ago)

Quote:

mogur said:
The reason H2O2 won't work is that complex carbs and enzymes inactivate it. The only way that I know to make grains compatible with peroxide is to break down the enzymes in grain by heat sterilization. If you just throw peroxide in with untreated grains, you will get a bunch of fizzing and no peroxide left. In theory there should be an amount of H2O2 that would break down the enzymes and have just enough left over to attack the contams and thereby sterilize the grain, but there's no known practical way to determine that precise amount, and any left over would attack the mushroom spores. Mycelium is self-protecting from peroxide, because it will produce enzymes that breakdown the peroxide in the vicinity of the mycelium, but spores don't have that protective mechanism available.

I don't know about bleach, but I assume it would not rinse off, and even if it did, the rinse would have to be sterile, so what is gained? Also, both H2O2 and bleach are not all that effective against endospores, so other problems aside, heat is both more effective and easier.




What about inoculating with LC?

I was considering trying a method where I would make a few half pint BRF jars (like 5, with multi spore inoculation). Then try soaking the grains 24+ hours to allow endospore germination. Then rinse very throughly and soak in H202. Then I would load into jars and microwave. Once cooled inside of a glove box (Or glove bag, seeing as I wanted this to be the economy tek.) I would crumble and transfer half of the fully colonized cake to each quart jar of grain. Leaving 10 quart jars fully colonized....Now that's plenty to start a large double or mono tub.

Basically my game plan was to out run the bacteria, anyone with G2G transfer experiance knows how fast they can be.

Maybe it would be easier to just make 20 BRF jars tho LOL.


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Edited by musher_420 (04/09/06 06:28 PM)


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Offlinemogur
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Re: Sterilzation of grain by soaking? [Re: FooMan]
    #5497373 - 04/09/06 06:23 PM (17 years, 9 months ago)

Oh yeah, FooManShroom reminded me that the grain makes a huge difference. I wouldn't even mess with non-PC methods with grains like rye, barley, or hard red wheat. They're great grains for colonization, but usually rife with endospores, so they're the toughest of all to sterilize. Popcorn, WBS, and soft white wheat are much more suitable for steaming or microwaving. And as FooMan says, BRF in the PF tek is the best of all for simple steaming.


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Invisibletrauma47645
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Re: Sterilzation of grain by soaking? [Re: musher_420]
    #5497385 - 04/09/06 06:27 PM (17 years, 9 months ago)

but spawning BRF to grain kinda defeats the purpose you might as while spawn the BRF directly to poo for a bulk grow if thats the case. Im trying to develope a non PC method for multispore inoculation of grain.. Right now the bleach or microwave option seem the most viable.. i think i am gonna PC 2 jars of WBS tonight as controls then do two WBS jars in the microwave and two WBS jars with bleach to see which works better if at all.. anyone else want to do the same thing would be a great help the more people involved in developing this tek will help to bring any error to light and would recieve equal due credit


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Invisiblemusher_420
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Re: Sterilzation of grain by soaking? [Re: trauma47645]
    #5497398 - 04/09/06 06:31 PM (17 years, 9 months ago)

I see where your coming from. And yes making BRF jars first and then spawning them does seem like a lengthy process. Basically I just wanted to find a way to expand the amount of spawn your working with. Because steaming 20 jars would be a bitch. Just doing like 5 and then microwaving 10 grain jars seems easier to me.... but I'm skeptical if it would work.


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Invisibletrauma47645
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Re: Sterilzation of grain by soaking? [Re: trauma47645]
    #5497402 - 04/09/06 06:31 PM (17 years, 9 months ago)

I really would like to get some really respected veteran growers in on this experiment if not i would really like some more of thier input (you reading this RR or roadkill) thanks foomanshroom if you have anymore idea it would really help.. but i dont think grinding the grain down like BRF would be the solution.. I am looking at this so closely because of the multiple inoculation points with grain.


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OfflineIGnosticAbhorI
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Re: Sterilzation of grain by soaking? [Re: trauma47645]
    #5497404 - 04/09/06 06:31 PM (17 years, 9 months ago)

PM Blackout, or look at his main threads.

He tried all the things you have mentioned, and more I'm sure.

-Gnostic


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Re: Sterilzation of grain by soaking? [Re: IGnosticAbhorI]
    #5499267 - 04/10/06 09:38 AM (17 years, 9 months ago)

Well worth experimenting with. I plan on doing more myself very soon.
http://www.shroomery.org/forums/showflat.php?Cat=0&Board=Forum2&Number=4323776

I have sterilized grains in single heatings in a microwave and am fairly certain it is possible to repeat with normal steaming. I would soak the grains for a few days, now the endospores have germinated so you can now add bleach, this means the bleach is not degraded by the time the endospores germinate. Leave them a few days and you could add more bleach in case the first lot has degraded. Now you could rinse off the bleach and salts and steam the grains.

The ulimate goal would be soaking, then adding bleach and not heating at all, just allow the bleach to degrade in the sunlight.


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InvisibleFooMan
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Re: Sterilzation of grain by soaking? [Re: blackout]
    #5499368 - 04/10/06 10:32 AM (17 years, 9 months ago)

Quote:

blackout said:
Well worth experimenting with. I plan on doing more myself very soon.
http://www.shroomery.org/forums/showflat.php?Cat=0&Board=Forum2&Number=4323776

I have sterilized grains in single heatings in a microwave and am fairly certain it is possible to repeat with normal steaming. I would soak the grains for a few days, now the endospores have germinated so you can now add bleach, this means the bleach is not degraded by the time the endospores germinate. Leave them a few days and you could add more bleach in case the first lot has degraded. Now you could rinse off the bleach and salts and steam the grains.

The ulimate goal would be soaking, then adding bleach and not heating at all, just allow the bleach to degrade in the sunlight.




Now that's smart thinking! :thumbup:


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