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OfflineBlake_Shroom
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Re: On endospores [Re: Novanity1]
    #18810075 - 09/07/13 12:17 AM (10 years, 4 months ago)

I feel like i was debating your room mate instead of you :rasta:

Anyways, let him call you a fool. Then munch on your fungus all by your self and laugh at him :africaface:


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Offlinekrypto2000
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Re: On endospores [Re: Blake_Shroom]
    #18810095 - 09/07/13 12:33 AM (10 years, 4 months ago)

Bacterial contaminants usually will show up on agar as wet spots. If your agar is too dry then they're not even going to germinate so it's not an issue.


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OfflineHobart Cutter
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Re: On endospores [Re: krypto2000]
    #18810424 - 09/07/13 03:46 AM (10 years, 4 months ago)

I was under the impression that endospores in grains form because of the lack of humidity. They are in other words there already, in abundance even, in some grains.
This is why a 12 hr wait between soaking and cooking improves the results.

Secondly, many of us use PCs that do not make it above 15psi. Many hover around 13 or 14, even if advertized as doing 15. (I believe that crossing the 15psi line is even illegal in consumer devices.)

And many of us also use PCs with a declared lower pressure.

All in all, in practical terms some slight traces of endospores is a reality that is hard to get away from, at least for the small scale hobbyist.


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OfflineHobart Cutter
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Re: On endospores [Re: Novanity1]
    #18810433 - 09/07/13 03:52 AM (10 years, 4 months ago)

Quote:

Novanity1 said:
I don't think the "race" is due to the sterilization procedures as much as it is the inoculation. Think about it. Spores are obtained from mushrooms that were exposed to open air. That's why we use agar to provide a clean culture. But this is where human error comes into play I suppose. There could very well be bacteria latent in a visually clean culture.




The problem with this line of thinking is that bacteria can't outpace mycelium when it comes to spreading. If you have bacteria in the syringe, it will either take off right away and prevent the mycelium from establishing, or be over run.

Endospore problems usually manifest later on in the process, in places far from the inoculation points, where the mycelium has yet to reach.


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InvisibleSpitballJedi
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Re: On endospores [Re: elasticaltiger]
    #18837969 - 09/13/13 06:00 PM (10 years, 4 months ago)

This conversation has come up a few times lately.

My understanding so far:

Although autoclaving kills endospores, it is unlikely that it kills absolutely 100% in our grain jars.

But, if we reactivate them to there normal state, then they are no longer an issue once we start sterilizing.

If only a minimal number of endospores remain after the reactivation process, then that leaves an even smaller likelihood that any will survive the PC. Some endospores take longer to reactivate in the conditions we generally provide, ie soaking in water.

So in retrospect, "racing against contams" might be a bit of a misnomer. It would be like Jesse Owens racing against my 2 month old niece.

On another note...I'm no biologist or anything, but I would think endospore producers are more common than your friend let's on to be. Bacillus comes to mind. Isn't that fairly common? Isn't it a common grain spawn contam?

In my own experience and some testing, I have concluded that if you follow common and tested and proven methods for sterilizing grains and you get a contam in your jar, it will be either your inoculate or technique.

Over the weekend, I will PC some grains and water without soaking. I will let them sit on the shelf and see if they grow anything on their own.


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OfflineLTSwoomz
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Re: On endospores [Re: Blake_Shroom]
    #18838031 - 09/13/13 06:14 PM (10 years, 4 months ago)

Quote:

Blake_Shroom said:
I feel like i was debating your room mate instead of you :rasta:

Anyways, let him call you a fool. Then munch on your fungus all by your self and laugh at him :africaface:




>.<

I feel like a buncha you are high as fuck and forgot to read the first and last bits of his post which give it some context.

He didn't call him a fool. The point of the post is that his room mate, who is a biochemist, explained why you shouldn't be worrying about anything in your jars after PCing because PCing effectively murders 99% of the things that could be present there under almost all normal circumstances. He also explains that endospores, which are a cornerstone of one side of the topic, are not given reasonable time to be created, in the rare but possible chance that your jars are exposed to a bacteria that can create endospores (read: most bacteria can't).


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OfflineStromriderM
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Re: On endospores [Re: SpitballJedi]
    #18838039 - 09/13/13 06:15 PM (10 years, 4 months ago)

Quote:

Spitballjedi Said:

Over the weekend, I will PC some grains and water without soaking. I will let them sit on the shelf and see if they grow anything on their own.




I am curious to see what becomes of this. Please do a write up for us.  :thanx:


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InvisibleSpitballJedi
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Re: On endospores [Re: Stromrider]
    #18838099 - 09/13/13 06:31 PM (10 years, 4 months ago)

Quote:

Hobart Cutter said:
Quote:

Novanity1 said:
I don't think the "race" is due to the sterilization procedures as much as it is the inoculation. Think about it. Spores are obtained from mushrooms that were exposed to open air. That's why we use agar to provide a clean culture. But this is where human error comes into play I suppose. There could very well be bacteria latent in a visually clean culture.




The problem with this line of thinking is that bacteria can't outpace mycelium when it comes to spreading. If you have bacteria in the syringe, it will either take off right away and prevent the mycelium from establishing, or be over run.

Endospore problems usually manifest later on in the process, in places far from the inoculation points, where the mycelium has yet to reach.




The problem with that line of thinking is I see pics all the time where bacteria has taken over a jar, cake, or tub of healthy mycelium. I personally have experienced it as well.

Endospores are formed by bacteria. When the endospore reactivates, it is simply reverting back to its original bacterial state and can overtake your grow.

Poor sterilization procedure can leave bacteria or their endospores alive and can come back to bite you later.


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InvisibleSpitballJedi
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Re: On endospores [Re: Stromrider]
    #18838103 - 09/13/13 06:31 PM (10 years, 4 months ago)

Quote:

psillyshroomer said:
Quote:

Spitballjedi Said:

Over the weekend, I will PC some grains and water without soaking. I will let them sit on the shelf and see if they grow anything on their own.




I am curious to see what becomes of this. Please do a write up for us.  :thanx:




I will document every detail I can think of.


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OfflineStromriderM
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Re: On endospores [Re: SpitballJedi]
    #18838108 - 09/13/13 06:32 PM (10 years, 4 months ago)

:thumbup:


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OfflineTrippy_Penguin
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Re: On endospores [Re: LTSwoomz]
    #18838122 - 09/13/13 06:37 PM (10 years, 4 months ago)

Quote:

LTSwoomz said:
He also explains that endospores, which are a cornerstone of one side of the topic, are not given reasonable time to be created, in the rare but possible chance that your jars are exposed to a bacteria that can create endospores (read: most bacteria can't).




Endospores are not going to be created during the cultivation process. Endospores are created to survive in conditions that would otherwise not allow for the species to survive.


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InvisibleSpitballJedi
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Re: On endospores [Re: elasticaltiger]
    #18838167 - 09/13/13 06:47 PM (10 years, 4 months ago)

Quote:

elasticaltiger said:
5)  In the off chance that your substrate IS contaminated with one of the FEW kinds of bacteria that can 'hibernate' as an endospore and  you are heating your pressure cooker on the stoves 'high' setting bringing the temperature and pressure up quickly the bacteria will most likely never even have the time necessary to form an endospore before the whole cell is destroyed out-right by the heat.




If, as I implied earlier, endospore producing bateria and/or their endospores are indeed more common than your friend infers, then it might mean it's very likely endospores are already on my grains when I get them.

Rye grains containing endospores is something commonly said on these forums and is part of the reason for the soak.

If my claim is true, then "5)" is a mute point.

I'm just playing Devil's Advocate here. I can't claim to know for sure.


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OfflineTheStormsEye
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Re: On endospores [Re: SpitballJedi]
    #18838280 - 09/13/13 07:12 PM (10 years, 4 months ago)

:popcorn:


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Invisiblecynical bastad
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Re: On endospores [Re: TheStormsEye]
    #18838407 - 09/13/13 07:41 PM (10 years, 4 months ago)

I knew/know very little, but i thought i must have gotten some really clean grains a while back or i did an awesome job at washing my grains...like i wash muh sticky rice.  I had washed them for a couple hours untill the water ran clear, but a friend told me i had messed up by not soaking them to open up the endospores and make them vulnerable to the heat in the pc.  I had made up 36 jars for G2G and only PC'd them, out of those only 2 contaminated... I may have gotten really lucky still, i dunno.  I didn't know you were supposed to soak.


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OfflineRogerRabbitM
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Re: On endospores [Re: elasticaltiger]
    #18838544 - 09/13/13 08:13 PM (10 years, 4 months ago)

Quote:

elasticaltiger said:
My roommate is a bio chemist. . .

After hearing him laugh his ass off at me and then proceeding to lecture me this is what I have walked away with. . .



5)  In the off chance that your substrate IS contaminated with one of the FEW kinds of bacteria that can 'hibernate' as an endospore and  you are heating your pressure cooker on the stoves 'high' setting bringing the temperature and pressure up quickly the bacteria will most likely never even have the time necessary to form an endospore before the whole cell is destroyed out-right by the heat.

So there is no 'race' because you jars PROBABLY are going to be sterile assuming you pressure cooked it for a while and brought the heat up quickly.

Any other contamination is due to human error.




That laughing sound was actually me rofl at any so-called bio chemist who understands so little about bio chemistry as to utter that line of total bullshit about bacterial endospores needing to form in the pressure cooker.

Endospore forming bacteria are not rare at all, but common on all cereal grains, such as the millet, rye, wheat berries, etc., used by mushroom growers.  Furthermore, sterilizing a jar of grains is far different than sterilizing a scalpel or other tool.  The steam doesn't penetrate the jar, but rather it heats up by conduction through the glass, which is an insulator.  Combine that with grains with spaces between them and they don't heat up very fast. 

This is why we recommend 90 minutes for quart jars of grains.  This ensures the grains in the very center of the jar are exposed to at least 20 minutes at 121C.  This will kill pre-moistened bacterial endospores, whether they germinated in the soak water or not.

Oh, and for the record, tell your friend endospores form when the grains are still on the stalk in the field.
RR


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InvisibleFrankHorrigan
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Re: On endospores [Re: RogerRabbit]
    #18838953 - 09/13/13 09:29 PM (10 years, 4 months ago)

:smilingpuppy:

That was fantastic.

Quote:

RogerRabbit said:
This is why we recommend 90 minutes for quart jars of grains.  This ensures the grains in the very center of the jar are exposed to at least 20 minutes at 121C.  This will kill pre-moistened bacterial endospores, whether they germinated in the soak water or not.





This explains a lot of things I was unsure of :thumbup:


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Invisibleelasticaltiger
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Re: On endospores [Re: RogerRabbit]
    #18839420 - 09/13/13 11:40 PM (10 years, 4 months ago)

Quote:

RogerRabbit said:
Quote:

elasticaltiger said:
My roommate is a bio chemist. . .

After hearing him laugh his ass off at me and then proceeding to lecture me this is what I have walked away with. . .



5)  In the off chance that your substrate IS contaminated with one of the FEW kinds of bacteria that can 'hibernate' as an endospore and  you are heating your pressure cooker on the stoves 'high' setting bringing the temperature and pressure up quickly the bacteria will most likely never even have the time necessary to form an endospore before the whole cell is destroyed out-right by the heat.

So there is no 'race' because you jars PROBABLY are going to be sterile assuming you pressure cooked it for a while and brought the heat up quickly.

Any other contamination is due to human error.




That laughing sound was actually me rofl at any so-called bio chemist who understands so little about bio chemistry as to utter that line of total bullshit about bacterial endospores needing to form in the pressure cooker.

Endospore forming bacteria are not rare at all, but common on all cereal grains, such as the millet, rye, wheat berries, etc., used by mushroom growers.  Furthermore, sterilizing a jar of grains is far different than sterilizing a scalpel or other tool.  The steam doesn't penetrate the jar, but rather it heats up by conduction through the glass, which is an insulator.  Combine that with grains with spaces between them and they don't heat up very fast. 

This is why we recommend 90 minutes for quart jars of grains.  This ensures the grains in the very center of the jar are exposed to at least 20 minutes at 121C.  This will kill pre-moistened bacterial endospores, whether they germinated in the soak water or not.

Oh, and for the record, tell your friend endospores form when the grains are still on the stalk in the field.
RR




I showed him this and he just said 'yeah well that's just your jar's environment with grains and stuff' then went to his room to blaze.

Is it that really different for other environments RR?  I mean, outside of pressure cooking grains? Is there like, a 'mean / mode amount' of endospores all around us on anything exposed to open air or is it particular to grains / plant matter?

Thank you for setting the record straight.  I'm kinda sorry I ever even related what he had to say now.  Request for mods to delete thread so people don't get the wrong info in the future. /Embarrassment. /Humiliation.


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Re: On endospores [Re: elasticaltiger]
    #18839916 - 09/14/13 02:47 AM (10 years, 4 months ago)

Deleting this thread is a terrible idea tiger! This discussion had produced some valuable information


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OfflineHobart Cutter
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Re: On endospores [Re: SpitballJedi]
    #18840008 - 09/14/13 03:36 AM (10 years, 4 months ago)

Quote:

SpitballJedi said:
Quote:

Hobart Cutter said:
Quote:

Novanity1 said:
I don't think the "race" is due to the sterilization procedures as much as it is the inoculation. Think about it. Spores are obtained from mushrooms that were exposed to open air. That's why we use agar to provide a clean culture. But this is where human error comes into play I suppose. There could very well be bacteria latent in a visually clean culture.




The problem with this line of thinking is that bacteria can't outpace mycelium when it comes to spreading. If you have bacteria in the syringe, it will either take off right away and prevent the mycelium from establishing, or be over run.

Endospore problems usually manifest later on in the process, in places far from the inoculation points, where the mycelium has yet to reach.




The problem with that line of thinking is I see pics all the time where bacteria has taken over a jar, cake, or tub of healthy mycelium. I personally have experienced it as well.




And therefor it must be a bad syringe? A does not follow B here. And cakes and tubs are open to air and not really relevant to whether bad inoculant is the primary source of bacterial contams.

Quote:


Endospores are formed by bacteria. When the endospore reactivates, it is simply reverting back to its original bacterial state and can overtake your grow.




Sure, but spore solution is liquid. There should not be a lot of endospores in there even if it's contaminated. Bacterial growth should therefore start quickly, and also be vulnerable to being digested by the mycelium.


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Re: On endospores [Re: RogerRabbit]
    #18842257 - 09/14/13 07:59 PM (10 years, 4 months ago)

Quote:

RogerRabbit said:
This is why we recommend 90 minutes for quart jars of grains.  This ensures the grains in the very center of the jar are exposed to at least 20 minutes at 121C.  This will kill pre-moistened bacterial endospores, whether they germinated in the soak water or not.
RR




http://www.shroomery.org/forums/showflat.php/Number/15294337#15294337
Quote:

RogerRabbit said:
The idea that soaking causes the endospores to germinate so we can 'kill' them in the PC is outdated, and more recent research is proving that theory to be flawed.

The fact is, if you soak too long, you end up with more endospores than you started with, since many endospore forming bacteria begin to produce new endospores within hours of germination.
RR




http://www.shroomery.org/forums/showflat.php/Number/17034012#17034012
Quote:

RogerRabbit said:
Don't get hung up on the endospore problem.  Soak in hot water at least 4 hours, which will hydrate/soften the grains enough that they won't burst when you boil.  If the grains are hydrated, so are the bacterial endospores which will then be nuked in the pressure cooker.  As foo said, long soaks can be counterproductive because many endospores will germinate and then make more endospores right in the soak water.  You could easily end up with more than you started with.  Consider 24 hours maximum.
RR




These 2 quotes seem to suggest forming more endospores from soaking too long is a bad thing. But I could also interpret it as saying it don't matter because they will be killed too.

Are you telling us there should not be any contamination to "race against" as long as we properly sterilize? That this "race against contams" is an outdated belief?

The reason I ask is so I can be better informed when trying to help others with contam issues.


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