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OfflineShadow of magus
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Moisture accumulation in jars
    #23798395 - 11/03/16 05:15 PM (7 years, 2 months ago)

Hello again to the great community of shroomery,

Ive been searching this site for quite sometime now prior to me even becoming a member and I just want to say I'm honored be here with this great community.

I've been encountering a problem for quite sometime now and through all my research, I have been left with more questions than answers. I've been having a problem for quite sometime now with a lot of runs on grain spawn, the accumulation of moisture.

I used a grain spawn tek by Mark Kieth who's video is right on you tube which displayed the grain in a 24 hour soak with two tablespoons of gypsum, followed by a 20 minute boil. Drain, let dry, place in jars. Now before I placed them in jars I make sure they are completely dry by letting them sit on some tissue paper to see the paper absorbs any water, they were completely dry.

I created injection ports on the lids by drilling a hole for inoculation, and one for air exchange using polyfyl filters.

All seemed to be going well first few weeks, they were colonizing at a decent rate, then they stopped and I actually saw water in jars. This definitely brought them to a hault, I don't see any contamination though. I feel like the grain are absorbing too much water and some how release it after a few weeks but I will provide pics to show what I mean. I'm thinking I'm boiling for too long.
Jar1
Jar 1

Jar 2


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OfflineGreg
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Re: Moisture accumulation in jars [Re: Shadow of magus]
    #23798398 - 11/03/16 05:17 PM (7 years, 2 months ago)

Damn those are some nasty jars.
Grain prep isn't your problem here.
That's bacteria halting your colonization and causing all of the water/metabolites...

What did you inoculate them with?
Did you inoculate them in open air?


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Re: Moisture accumulation in jars [Re: Greg]
    #23798499 - 11/03/16 05:47 PM (7 years, 2 months ago)

that shit yo lip has some shit on its lip son. :niggawhy:

something else is at play here. usually if i have water in the jars
its my prep that got fucked up.

honestly some people are great with grains. ive done maybe 100+ quarts
in the last three months and ive had to ditch a few cause i fucked up.
granted you said you checked if they were dry but unless you inoculated
while taking a dump.

or did you use a dirty LC or something..??


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OfflineShadow of magus
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Re: Moisture accumulation in jars [Re: Greg]
    #23798551 - 11/03/16 06:06 PM (7 years, 2 months ago)

I don't have a flow hood. I inoculated in a small room where I sprayed anti bacterial stuff and even isopropyl alcohol and I let it sit.


Damn, do you think they're lost?


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InvisibleMad Season
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Re: Moisture accumulation in jars [Re: Shadow of magus]
    #23798560 - 11/03/16 06:08 PM (7 years, 2 months ago)

Still air box FTW


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OfflineGreg
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Re: Moisture accumulation in jars [Re: Mad Season]
    #23798566 - 11/03/16 06:11 PM (7 years, 2 months ago)

I sure wouldn't use those grains for anything other than a trash can filler.
As Mad said, use a SAB: https://www.shroomery.org/forums/showflat.php/Number/20048771#20048771


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Re: Moisture accumulation in jars [Re: Greg] * 1
    #23798571 - 11/03/16 06:11 PM (7 years, 2 months ago)

Quote:

Shadow of magus said:
Hello again to the great community of shroomery,

Ive been searching this site for quite sometime now prior to me even becoming a member and I just want to say I'm honored be here with this great community.

I've been encountering a problem for quite sometime now and through all my research, I have been left with more questions than answers. I've been having a problem for quite sometime now with a lot of runs on grain spawn, the accumulation of moisture.

I used a grain spawn tek by Mark Kieth who's video is right on you tube which displayed the grain in a 24 hour soak with two tablespoons of gypsum, followed by a 20 minute boil. Drain, let dry, place in jars. Now before I placed them in jars I make sure they are completely dry by letting them sit on some tissue paper to see the paper absorbs any water, they were completely dry.

I created injection ports on the lids by drilling a hole for inoculation, and one for air exchange using polyfyl filters.

All seemed to be going well first few weeks, they were colonizing at a decent rate, then they stopped and I actually saw water in jars. This definitely brought them to a hault, I don't see any contamination though. I feel like the grain are absorbing too much water and some how release it after a few weeks but I will provide pics to show what I mean. I'm thinking I'm boiling for too long.
Jar1
Jar 1

Jar 2





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OfflineGreg
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Re: Moisture accumulation in jars [Re: azur]
    #23798575 - 11/03/16 06:12 PM (7 years, 2 months ago)

Azur you've sure been putting that gif to good use lately :lol:


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Re: Moisture accumulation in jars [Re: Greg]
    #23798585 - 11/03/16 06:16 PM (7 years, 2 months ago)

Just twice so far


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OfflineGreg
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Re: Moisture accumulation in jars [Re: azur]
    #23798601 - 11/03/16 06:22 PM (7 years, 2 months ago)

Keep at it then :hehehe:


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Re: Moisture accumulation in jars [Re: azur]
    #23798604 - 11/03/16 06:22 PM (7 years, 2 months ago)

OP, did you sterilize them?  I'm guessing you did or they'd be much worse off...was curious as you didn't note that step.  Also, how old are they?  Again, curious as you mention weeks... seems long even for ms straight to grain...or not.


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Re: Moisture accumulation in jars [Re: Shadow of magus]
    #23798615 - 11/03/16 06:26 PM (7 years, 2 months ago)

Quote:

Shadow of magus said:
I don't have a flow hood. I inoculated in a small room where I sprayed anti bacterial stuff and even isopropyl alcohol and I let it sit.


Damn, do you think they're lost?




no flaming of needle? weak bruh

toss and redo. no biggie. live and learn

build a still air box. best thing you can own..besides a pc or dehydrator.


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OfflineShadow of magus
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Re: Moisture accumulation in jars [Re: mushboy]
    #23798783 - 11/03/16 07:17 PM (7 years, 2 months ago)

Quote:

mushboy said:
Quote:

Shadow of magus said:
I don't have a flow hood. I inoculated in a small room where I sprayed anti bacterial stuff and even isopropyl alcohol and I let it sit.


Damn, do you think they're lost?




no flaming of needle? weak bruh

toss and redo. no biggie. live and learn

build a still air box. best thing you can own..besides a pc or dehydrator.



Quote:

mushboy said:
Quote:

Shadow of magus said:
I don't have a flow hood. I inoculated in a small room where I sprayed anti bacterial stuff and even isopropyl alcohol and I let it sit.


Damn, do you think they're lost?




no flaming of needle? weak bruh

toss and redo. no biggie. live and learn

build a still air box. best thing you can own..besides a pc or dehydrator.




I definitely sterilized for 15 psi for one hour and I torched the needle upon inoculation, it's been about 4 weeks.

So I should toss these? Is it ok if I re use jars after boiling them for 1.5 hours?


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Re: Moisture accumulation in jars [Re: Shadow of magus]
    #23799149 - 11/03/16 09:25 PM (7 years, 2 months ago)

Just wash them and reuse.


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Re: Moisture accumulation in jars [Re: Mycolorado]
    #23799183 - 11/03/16 09:42 PM (7 years, 2 months ago)

Eh, if you're PC-ing again anyway, I usually don't even bother to wash my jars. Had a bad batch of oysters contam and I left them alone for six months just to see what would happen. Was nasty (fruit loops+poop smell) when I finally tossed the grains. Soaked new grains in the jars for 48 hours without washing, PC'ed, and now I have happy and clean mycelium munching on some WBS.

If anything, I figure the dead bacterial leftovers will be more nutrients.


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Re: Moisture accumulation in jars [Re: Kryptos]
    #23799225 - 11/03/16 09:56 PM (7 years, 2 months ago)

I hate this kind of shitty advice. 
And I know most the guys on here are lazy hippies and do the same, but that doesn't make it right.
When we sterilize grain, we give ourselves a window of opportunity because we cannot kill everything. Therefore, we should wipe our asses as clean as possible.


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FALL IN LOVE WITH LC
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OfflineShadow of magus
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Re: Moisture accumulation in jars [Re: azur]
    #23799771 - 11/04/16 05:23 AM (7 years, 2 months ago)

Thanks guys, hopefully I can redeem myself to you guys and myself with a good successful batch.


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OfflineKryptos
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Re: Moisture accumulation in jars [Re: azur]
    #23801260 - 11/04/16 05:21 PM (7 years, 2 months ago)

Quote:

azur said:
I hate this kind of shitty advice. 
And I know most the guys on here are lazy hippies and do the same, but that doesn't make it right.
When we sterilize grain, we give ourselves a window of opportunity because we cannot kill everything. Therefore, we should wipe our asses as clean as possible.




I guess this depends on sterilization procedures. I'm fairly certain you're more skilled than I am, since I've followed your advice in the past, but at the same time, my tester jar (First thing I PCed when I got my presto ~18 months back) is still completely free of any kind of growth or contamination. It was allowed to ferment for 72 hours, and I added some yard dirt to it before PC cycle to make it extra contaminated. I'm pretty confident that my PC cycle kills *everything* as there has been absolutely no change in that jar over a period longer than a year.

My PC cycle, for reference: 15 psi, one and a half hours, timer starts when PC reaches pressure and heat is turned off when timer ends. PC is left to cool on the same stovetop where it was heated, so it ends up staying at 15 psi for 1:45-2 hours.


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Re: Moisture accumulation in jars [Re: Kryptos]
    #23801305 - 11/04/16 05:35 PM (7 years, 2 months ago)

If there's any endospores in your grains, they're not truly sterile with an 18 psi and 1.45-2 hr cycle. I wonder what your 1 year old jar smells like.. if it's all good, thank your source for not using endospore pesticides, not your PC cycle


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Re: Moisture accumulation in jars [Re: Mad Season]
    #23801549 - 11/04/16 06:58 PM (7 years, 2 months ago)

Endospores are killed in 15m at 15 psi in aqueous media. But in grains or bulk matter you almost never achieve complete sterilization. Contamination may not germinate right away or ever or until some conditions are met. But don't think that 15 psi 90 minutes sterilized grains completely


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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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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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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.


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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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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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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
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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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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.


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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
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OfflineShadow of magus
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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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