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Offlineshellzenone
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repetative contaminations
    #21831091 - 06/20/15 02:02 AM (8 years, 7 months ago)

I have been working with a single master culture grown on wbs. All jars look and smell contam free, yet I haven't had a successful grow in over 4 months. At first I was using a coir,verm,coffee and gypsum substrate and I began to attribute the contaminations to the coffee, so I dropped it from the mix. Still my last attempts have resulted in a trich contamination... Is it possible that my master culture is contaminated? Again, the jars never show signs of contamination. I usually pasteurize using the bucket tech, only I use my PC in place of the bucket. I've read that a properly pasteurized substrate will not contaminate unless the spawn is contaminated. Anyone ever have a properly pasteurized coir and verm substrate get contaminated? I can't figure it out. I've had trich in spawn jars before, and it always shows. These jars never show any signs!!! I plan on pasteurizing some coir verm and gypsum mix and leaving it out to see if anything grows on it. Maybe its my pasteurization method I need to fix? Anyway, I'll post results on my experiment. Just want to know what you guys think.


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Invisibleoontribe
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Re: repetative contaminations [Re: shellzenone]
    #21831383 - 06/20/15 06:00 AM (8 years, 7 months ago)

iam not experienced, and it could be too much water in the sub or room temp.


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OfflineGreenRabbit
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Re: repetative contaminations [Re: shellzenone]
    #21831625 - 06/20/15 08:16 AM (8 years, 7 months ago)

Quote:

shellzenone said:
Is it possible that my master culture is contaminated?





It's more than likely..


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Invisiblemaddchef
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Re: repetative contaminations [Re: GreenRabbit]
    #21831790 - 06/20/15 09:31 AM (8 years, 7 months ago)

Are you seeing any bacteria? I recently had an issue with wbs where no matter how long I sterilized I would have endospores germinate and damage the mycelium enough to allow mold to set in. Try prepping your grain extra dry next run and see if it helps any of switch grains.


--------------------
In the land of the blind, the one eyed man is king.

All mushrooms are edible, but some only once.....                     

                                        Easier than cakes

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Offlineshellzenone
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Re: repetative contaminations [Re: maddchef]
    #21832881 - 06/20/15 03:59 PM (8 years, 7 months ago)

No, I don't think so. Usually bacteria gives off a smell. I see no signs of contamination.


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OfflineFunnyLight
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Re: repetative contaminations [Re: shellzenone]
    #21832923 - 06/20/15 04:14 PM (8 years, 7 months ago)

Maybe do a 50/50 run of pasteurized grains vs. sterilized grains.

I always sterilized my grains. Specially WBS, they've got so much stuff tucked up in there.

Time to get into some Agar and grow out your master culture to see if any baddies pop up


--------------------
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Offlineshellzenone
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Re: repetative contaminations [Re: FunnyLight]
    #21833004 - 06/20/15 04:38 PM (8 years, 7 months ago)

I've never heard of pasteurizing grain?! And yes, I'll have to test my master culture.


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OfflineMachiavelliavore
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Re: repetative contaminations [Re: shellzenone]
    #21833074 - 06/20/15 05:01 PM (8 years, 7 months ago)

I had some sort of milky mold contaminate one of my master cultures.  It was hard to see and not very prolific at the beginning, but it took over the whole jars eventually.  The jars smelled somewhat tangy.

Btw I've only sterilized my WBS for 65 minutes, well bellow what's usually suggested, and had jars sitting for 4 weeks with no signs of contamination.


--------------------


I spawned some popcorn casings and had double-overlay cause I didn't put enough hydrogen peroxide in my automated aquarium mister.  I only got one mushroom so I cut off the head part where the seeds fall from and put it in a jar of LC and sprayed it all over a tin of PF cakes I made with gravel, cardboard, and bisquick in my microwave.  I think it will be good cause B+ is so potent.
Triggered yet?

Only a square would say "a cube is a cube."


No, this does not look right...


Edited by Machiavelliavore (06/20/15 05:02 PM)


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Offlineshroomyaxn
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Re: repetative contaminations [Re: shellzenone]
    #21833103 - 06/20/15 05:08 PM (8 years, 7 months ago)

Quote:

shellzenone said:
I've read that a properly pasteurized substrate will not contaminate unless the spawn is contaminated. Anyone ever have a properly pasteurized coir and verm substrate get contaminated?




You've read correctly.

How do you pasteurize your substrate mix?


Edited by shroomyaxn (06/20/15 06:45 PM)


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OfflineRyeJar
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Re: repetative contaminations [Re: shroomyaxn]
    #21833180 - 06/20/15 05:33 PM (8 years, 7 months ago)

This is a thread for mad season to chime in with his expertise with bac on grain


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Offlineshellzenone
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Re: repetative contaminations [Re: shroomyaxn]
    #21833194 - 06/20/15 05:38 PM (8 years, 7 months ago)

My pasteurization: I put my pc(23 qt) half filled with tap water on the stove for until it reaches about 130 Fahrenheit. Throw in a brick of coir, verm, gypsum, and mix it. Then I let the temp rise to 150 (I use a digital therm). Then I cover with the PC lid, turn off the heat and let it cool over night.

I've had success in the past using this method.


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Offlineshellzenone
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Re: repetative contaminations [Re: RyeJar]
    #21833202 - 06/20/15 05:39 PM (8 years, 7 months ago)

My recent contams have been trich, cobweb, and a black mold :sad:


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OfflineMachiavelliavore
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Re: repetative contaminations [Re: shellzenone]
    #21833246 - 06/20/15 05:52 PM (8 years, 7 months ago)

Not related to contamination, but having tried it in my PC and in a 30qt tote wrapped in foil and towels, I can say that it works much better to pour water ontop.

I think this is because the coir brick floats, so the bottoms swells immediately as you drop it in, and then the top doesn't hydrate without a good bit of mixing..  Maybe you don't have this issue since you heat it back up.


--------------------


I spawned some popcorn casings and had double-overlay cause I didn't put enough hydrogen peroxide in my automated aquarium mister.  I only got one mushroom so I cut off the head part where the seeds fall from and put it in a jar of LC and sprayed it all over a tin of PF cakes I made with gravel, cardboard, and bisquick in my microwave.  I think it will be good cause B+ is so potent.
Triggered yet?

Only a square would say "a cube is a cube."


No, this does not look right...


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Offlineshroomyaxn
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Re: repetative contaminations [Re: Machiavelliavore]
    #21833482 - 06/20/15 06:53 PM (8 years, 7 months ago)

This is how I go about pasteurizing.  I recently started using the autoclavable bags instead of the quart jars.  Primarily because the clean up is easier. 
Step by step:
•Load the substrate mix into the spawn bag.  Create a tightly packed rectangular cube containing the substrate or casing.  Secure the bag using autoclavable bag clamps, while squeezing out any trapped air. 
•Probe the bag with a thermometer so it reads the center of the substrate. 
•Place the bag in a pressure cooker.  Ensure there is a heat barrier between the spawn bag and the bottom of the pot.  The PC canning rack is best suited for a heat barrier. 
•Fill the PC with tap water so that the bags are almost entirely submerged.  A canning rack and a weight may be needed to keep the bag(s) from floating in the PC. 
•Turn on the stove bring the water temperature to a near boil, around 200+ degrees. 
•Turn off the stove once the water begins to steam then quickly seal the pressure cooker. 
•Usually within 45 minutes the interior bag temperature will have reached 140 degrees. 
    oThis can take up to an hour depending upon the components in the bag
    oPasteurization begins at 140 degrees and becomes unproductive after 160-170 degrees. 
•Ensure the internal bag temperature has maintained 140-160 °range for at least 60 minutes. 
•The water can be removed from the pressure cooker after pasteurization has been completed to speed up the cooling process.


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Invisiblemaddchef
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Re: repetative contaminations [Re: shroomyaxn]
    #21833651 - 06/20/15 07:39 PM (8 years, 7 months ago)

If you're seeing trich, cobweb, and black mold its either your sterilization is insufficient or your technique is flawed.


--------------------
In the land of the blind, the one eyed man is king.

All mushrooms are edible, but some only once.....                     

                                        Easier than cakes

I do science and shit.

:havesomescience:


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InvisibleMad Season
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Re: repetative contaminations [Re: maddchef]
    #21833667 - 06/20/15 07:43 PM (8 years, 7 months ago)

Are you sure you aren't seeing bacteria? It doesn't have to smell for there to be some. Mushrooms have a pretty pungent odor. You're looking for zero wet grains against the side of the jar


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Edited by Mad Season (06/20/15 07:43 PM)


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OfflineRyeJar
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Re: repetative contaminations [Re: Mad Season]
    #21833706 - 06/20/15 07:55 PM (8 years, 7 months ago)

Quote:

RyeJar said:
This is a thread for mad season to chime in with his expertise with bac on grain



Quote:

Mad Season said:
Are you sure you aren't seeing bacteria? It doesn't have to smell for there to be some. Mushrooms have a pretty pungent odor. You're looking for zero wet grains against the side of the jar




:nicesmile:


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InvisibleMad Season
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Re: repetative contaminations [Re: RyeJar]
    #21833727 - 06/20/15 08:04 PM (8 years, 7 months ago)

:wink: oh rye! I'm not sure if we should be bragging I have bacterial experience LOL!

Also don't pasteurize grains... you'll get way more bacteria. If anything you should up your pc times by half an hour to reduce it.


--------------------
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 (06/20/15 08:07 PM)


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Offlineshellzenone
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Re: repetative contaminations [Re: Mad Season]
    #21833795 - 06/20/15 08:22 PM (8 years, 7 months ago)

Well I'm not gonna give up guys, I'm sure this is a problem that can be fixed. I appreciate all your input. I'm 80% sure the pasteurization is not the issue... But I'll try pasteurizing for longer. I plan to leave the stove on at medium low heat and stiring every 20 mins or so to ensure the Sub is evenly heated. Also I plan on leaving my jars to colonize for a few more days after colonization.

lets say my home was infested with mold spores. Is it possible to contaminate the grain while constructing the mono?


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InvisibleMad Season
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Re: repetative contaminations [Re: shellzenone]
    #21833834 - 06/20/15 08:31 PM (8 years, 7 months ago)

I find pasteurizing in jars and getting internal temps between 140-160f for 1 hour is best, anything more is more detrimental. Most mold spores, trich and cobby included die at 130f for 30 minutes. Increasing time will just kill more beneficial microbes.

Up your pc times on grains to 2 hours minimum, or increase pressure if you have a good pc. Also if you don't do agar, get into it, to ensure you have a clean inoculant. Like I said you want no wet grains on the side of the jar.

Quote:

Mad Season said:
Well there's a difference between colonized and cleanly colonized.

This is colonized:

Lots of members have noticed cobweb and trich from jars that look like this or better. Wet spots are bacteria. Not wet grains or wet spots. To colonize cleanly is the hardest. I had to change my pc times to 2 hours to reduce it.

This is (ALMOST) cleanly colonized, that's how hard it is to have it completely clean. This isn't even completely clean:


This is completely clean spawn:



Anyone who's done this for awhile will testify this is the hardest part of the hobby and is what will dictate the success or failure of bulk.




Notice there's no wet grains. Hope this helps.


--------------------
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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