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OfflineKryptos
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Re: Moisture accumulation in jars [Re: bodhisatta]
    #23803469 - 11/05/16 01:52 PM (7 years, 2 months ago)

I'm just adding my experience with sterilization. I've used both PC and a lab autoclave, and I've let jars sit for weeks before inoculation with both methods and no contaminants. I have personally seen no evidence that my sterilization procedure results in anything other than sterile grains. Heck, I've gone over sterilization procedures used in the microbiology labs on campus (for both liquid and dry media), and mine are more rigorous.

You mention that contamination may not germinate until some conditions are met, but these conditions are ideal for mycelium growth. I guess, sure, there may be some (for example) methanogen bacterial endospores sitting around waiting for some methane to appear, but since that doesn't really apply at all, who cares?

It's hard to convince me that I am wrong, when all the available evidence points to me being right. As soon as it fails, and the failure isn't a result of improper procedure, I'll change my tune. Scientific method and all. This has yet to happen over the last year and a half.


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OfflineShadow of magus
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Registered: 10/31/16
Posts: 78
Last seen: 3 years, 9 months
Re: Moisture accumulation in jars [Re: Kryptos]
    #23809556 - 11/07/16 12:21 PM (7 years, 2 months ago)

I know what the problem was also...

I used Panaleous Cay, and they love cow dung and casing.

Just started new batches with golden teachers purple Mystics and AA Albinos, keeping my fingers crossed


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InvisibleMad Season
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Registered: 09/16/12
Posts: 12,666
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Re: Moisture accumulation in jars [Re: Kryptos]
    #23809749 - 11/07/16 01:28 PM (7 years, 2 months ago)

Quote:

Kryptos said:
I'm just adding my experience with sterilization. I've used both PC and a lab autoclave, and I've let jars sit for weeks before inoculation with both methods and no contaminants. I have personally seen no evidence that my sterilization procedure results in anything other than sterile grains. Heck, I've gone over sterilization procedures used in the microbiology labs on campus (for both liquid and dry media), and mine are more rigorous.

You mention that contamination may not germinate until some conditions are met, but these conditions are ideal for mycelium growth. I guess, sure, there may be some (for example) methanogen bacterial endospores sitting around waiting for some methane to appear, but since that doesn't really apply at all, who cares?

It's hard to convince me that I am wrong, when all the available evidence points to me being right. As soon as it fails, and the failure isn't a result of improper procedure, I'll change my tune. Scientific method and all. This has yet to happen over the last year and a half.



Microbiology labs aren't holding onto media for long periods of time. Look at grain spawn suppliers if you want to see what sterilization looks like with grains. A double cycle of 4+ hours at over 60 psi is common to actually achieve sterile grains that can be shipped and stored for a long time, and even then there's still the odd CFU that gets through. This has been extensively researched on this site and grain suppliers for YEARS. Just do some searching. The way we use grains is always a race against the CFUs. Increasing PC times and pressure will lower the amount that gets through, but if there are endospores in the beginning, some will undoubtedly get through with our cycles.

Quote:

Shadow of magus said:
I know what the problem was also...

I used Panaleous Cay, and they love cow dung and casing.

Just started new batches with golden teachers purple Mystics and AA Albinos, keeping my fingers crossed



Yeah the CLEAN grain jars of pan cyan like this:



Get spawned to manure and a casing. They grow just fine in grains so that wasn't the problem.


--------------------
contam and car window art
How to shroomery like a pro! (Seriously, everyone read this!)
Improve your sterile techniques! (A comprehensive guide to agar)
Links upon links of literally EVERYTHING UP TO DATE

AMU Q&A
No trees were killed in the sending of this message. However, a large number of electrons were terribly inconvenienced.


Edited by Mad Season (11/07/16 01:36 PM)


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OfflineShadow of magus
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Registered: 10/31/16
Posts: 78
Last seen: 3 years, 9 months
Re: Moisture accumulation in jars [Re: Mad Season]
    #23810777 - 11/07/16 07:21 PM (7 years, 2 months ago)

Quote:

Mad Season said:
Quote:

Kryptos said:
I'm just adding my experience with sterilization. I've used both PC and a lab autoclave, and I've let jars sit for weeks before inoculation with both methods and no contaminants. I have personally seen no evidence that my sterilization procedure results in anything other than sterile grains. Heck, I've gone over sterilization procedures used in the microbiology labs on campus (for both liquid and dry media), and mine are more rigorous.

You mention that contamination may not germinate until some conditions are met, but these conditions are ideal for mycelium growth. I guess, sure, there may be some (for example) methanogen bacterial endospores sitting around waiting for some methane to appear, but since that doesn't really apply at all, who cares?

It's hard to convince me that I am wrong, when all the available evidence points to me being right. As soon as it fails, and the failure isn't a result of improper procedure, I'll change my tune. Scientific method and all. This has yet to happen over the last year and a half.



Microbiology labs aren't holding onto media for long periods of time. Look at grain spawn suppliers if you want to see what sterilization looks like with grains. A double cycle of 4+ hours at over 60 psi is common to actually achieve sterile grains that can be shipped and stored for a long time, and even then there's still the odd CFU that gets through. This has been extensively researched on this site and grain suppliers for YEARS. Just do some searching. The way we use grains is always a race against the CFUs. Increasing PC times and pressure will lower the amount that gets through, but if there are endospores in the beginning, some will undoubtedly get through with our cycles.

Quote:

Shadow of magus said:
I know what the problem was also...

I used Panaleous Cay, and they love cow dung and casing.

Just started new batches with golden teachers purple Mystics and AA Albinos, keeping my fingers crossed



Yeah the CLEAN grain jars of pan cyan like this:



Get spawned to manure and a casing. They grow just fine in grains so that wasn't the problem.



Quote:

Mad Season said:
Quote:

Kryptos said:
I'm just adding my experience with sterilization. I've used both PC and a lab autoclave, and I've let jars sit for weeks before inoculation with both methods and no contaminants. I have personally seen no evidence that my sterilization procedure results in anything other than sterile grains. Heck, I've gone over sterilization procedures used in the microbiology labs on campus (for both liquid and dry media), and mine are more rigorous.

You mention that contamination may not germinate until some conditions are met, but these conditions are ideal for mycelium growth. I guess, sure, there may be some (for example) methanogen bacterial endospores sitting around waiting for some methane to appear, but since that doesn't really apply at all, who cares?


Quote:

Shadow of magus said:
I know what the problem was also...

I used Panaleous Cay, and they love cow dung and casing.

Just started new batches with golden teachers purple Mystics and AA Albinos, keeping my fingers crossed



Yeah the CLEAN grain jars of pan cyan like this:



Get spawned to manure and a casing. They grow just fine in grains so that wasn't the problem.




Thanks for bitch slapping the noob, that's actually really good news because I still have some Pan Cay syringes. Would rye grain work for Azurescens?


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OfflineShadow of magus
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Registered: 10/31/16
Posts: 78
Last seen: 3 years, 9 months
Re: Moisture accumulation in jars [Re: Shadow of magus]
    #23810790 - 11/07/16 07:24 PM (7 years, 2 months ago)

I like how clean was in capital letters. I sprayed disinfectant, alcohol and bleach, wore gloves bathed them in alcohol....torched the syringe...sprayed right before each inoculation. Don't know where I went wrong but hopefully will be corrected with the new batches


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Invisiblemorty422
Wuba-luba-dub-dub!
I'm a teapot

Registered: 07/06/16
Posts: 988
Re: Moisture accumulation in jars [Re: Shadow of magus]
    #23810916 - 11/07/16 08:00 PM (7 years, 2 months ago)

Quote:

Shadow of magus said:
I like how clean was in capital letters. I sprayed disinfectant, alcohol and bleach, wore gloves bathed them in alcohol....torched the syringe...sprayed right before each inoculation. Don't know where I went wrong but hopefully will be corrected with the new batches




The term CLEAN grain spawn = jars of grain that have no bacteria or other contaminates present. You could have done all your work in an operating room, with a flow hood and as long as you're using a spore syringe-there will be a high chance of there being bacteria in your grain spawn.

MS syringes harbor bacteria... that's just the name of the game. If you want clean grain spawn - get into agar.
It's the only way to do it.

I've been there before - I was lucky and my syringe didn't harbor a lOT of bacteria so I was able to pull off a few tubs with it... as soon as i got clean grain spawn, my yields tripled.

:cheers:


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InvisibleMad Season
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Registered: 09/16/12
Posts: 12,666
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Re: Moisture accumulation in jars [Re: morty422]
    #23810989 - 11/07/16 08:23 PM (7 years, 2 months ago)

:whathesaid:

The jars I posted are healthy panaeolus mycelium with no wet spots of bacteria. That's why you failed because there's bacteria. Why you have bacteria.. idk.. endospores, bad techniques, contaminated syringe (which is highly likely with panaeolus prints) are all quite plausible.


--------------------
contam and car window art
How to shroomery like a pro! (Seriously, everyone read this!)
Improve your sterile techniques! (A comprehensive guide to agar)
Links upon links of literally EVERYTHING UP TO DATE

AMU Q&A
No trees were killed in the sending of this message. However, a large number of electrons were terribly inconvenienced.


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OfflineShadow of magus
Stranger
Registered: 10/31/16
Posts: 78
Last seen: 3 years, 9 months
Re: Moisture accumulation in jars [Re: Mad Season]
    #23813510 - 11/08/16 05:44 PM (7 years, 2 months ago)

Agar.... Great idea.

I want to thank everyone for replying, I'm so glad to be a part of this site with this amazing community. Thank you for all of your help.


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