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Amazon Shop: ½ Pint Jars, Agar, Rye Grain, Wild Bird Seed

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Offlinelemunhed
The hustler'shustler

Registered: 05/12/03
Posts: 775
Loc: Ur moms house
Last seen: 12 years, 3 months
Rye jars keep contaminating, whats wrong?!
    #4272548 - 06/08/05 06:02 PM (12 years, 5 months ago)

Hey guys. My buddy has recently been doing alot of work with rye, he loves it because of its versatility (spawn or casings) its rapid colonization, high tryptophan content and ease of preparation, but his damn jars keep contaminating. Let me tell you what he is doing so that you can give him some pointers.


To start with, my friend uses 1qt ball jars, with the metal lid band and tyvek filters which he makes from tyvek postal envelopes. He is using spore syringes from the hawks eye so it most likely isn't that. The first batch of jars, my buddy added 250g rye and 280ml water (as recomended at mushmush.nl), he didn't soak the grain because it says for rye you don't need to (also at mushmush). He PC'ed them for about 1 and 1/2 hours then let them cool. Inoculation is done on a sterilie surface, duct tape is used to cover the inoculation hole. The needle is sterilized with 70% isopropyl alcohol between every new jar. Out of this batch he lost 2 to contamination out of 5.


Thinking it might be endospores that caused the problem because he didn't soak the grains, on the next batch my friend used the same technique but added the water a day in advance and allowed to soak. All 5 of these jars were lost to contamination. Do you guys have any idea what's going on because my friend sure as hell doesn't.


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You're pompeius, aren't you? I'll make a pompeian of you unless you hold your toungue!

-Tiberius Claudius Nero


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Offlineselar_doir
boner sponge

Registered: 04/27/05
Posts: 40
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Re: Rye jars keep contaminating, whats wrong?! [Re: lemunhed]
    #4272554 - 06/08/05 06:03 PM (12 years, 5 months ago)

duct tape might be too permeable- try electrical tape.


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OfflineMitchnast
Trial by Madness
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Registered: 10/28/99
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Re: Rye jars keep contaminating, whats wrong?! [Re: selar_doir]
    #4272583 - 06/08/05 06:14 PM (12 years, 5 months ago)

use liquid culture. spore syringes take too long to germinate.


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Invisibleagar
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Registered: 11/21/04
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Re: Rye jars keep contaminating, whats wrong?! [Re: lemunhed]
    #4272652 - 06/08/05 06:32 PM (12 years, 5 months ago)

Try soaking the rye 24 hours, then steep/simmer in hot water until the rye is fully expanded (without many bursting). That will allow endospores to germinate.

Rinse, drain & load jars 2/3rds to 3/4 full.

(above is WBS - but same procedure)

PC @ 15 psi 90 minutes.
Shake jars when removed from PC.
Allow to cool to room temp's.

Wear hair covering, face mask & alc swab your hands, when you inoculate. Swab jar tops with 70% alc, just before inoculation.

When you inoculate, remove needle sheath & keep needle in alc soaked - wrap swab, for minimal exposure to open air.

Hold swab wrapped needle over inoculation hole, push needle through swab, through tyvek & inoculate. As you pull needle out, keep it wrapped in the swab.

Use alc swabbed scotch tape to cover inoculate hole.

Might help? Might not.
But, it is a very sterile procedure for inoculation.


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OfflinekronnyQ
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Re: Rye jars keep contaminating, whats wrong?! [Re: lemunhed]
    #4272678 - 06/08/05 06:37 PM (12 years, 5 months ago)

I've ditched Tyvek for polyfill puff lids, because of contams.

If you are using Tyvek without the metal lids (actual lid not the band), my understanding is that when you inoculate the tyvek gets pushed down then pops back up after syringe removal, creating a small amount of suction, outside air going into the hole.

i would try using metal lids next time or just switch to polyfill.


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OfflineChemical_Bliss
Officer of thelaw...
Registered: 03/18/05
Posts: 279
Last seen: 11 years, 7 months
Re: Rye jars keep contaminating, whats wrong?! [Re: agar]
    #4272679 - 06/08/05 06:37 PM (12 years, 5 months ago)

I had this problem with rye too, 24 hour soak and a simmer stopped it. I wish I could offer more advice. Rye, in my opinion, seems much harder to seterilize then most other substrates.


--------------------
'divine moments of truth, total and utter cosmic stuff...'
'be here now... i love everybody'


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OfflineMych
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Registered: 03/25/05
Posts: 415
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Re: Rye jars keep contaminating, whats wrong?! [Re: agar]
    #4272687 - 06/08/05 06:39 PM (12 years, 5 months ago)

Quote:

agar said:
Try soaking the rye 24 hours, then steep/simmer in hot water until the rye is fully expanded (without many bursting). That will allow endospores to germinate.

Rinse, drain & load jars 2/3rds to 3/4 full.

(above is WBS - but same procedure)

PC @ 15 psi 90 minutes.
Shake jars when removed from PC.
Allow to cool to room temp's.

Wear hair covering, face mask & alc swab your hands, when you inoculate. Swab jar tops with 70% alc, just before inoculation.

When you inoculate, remove needle sheath & keep needle in alc soaked - wrap swab, for minimal exposure to open air.

Hold swab wrapped needle over inoculation hole, push needle through swab, through tyvek & inoculate. As you pull needle out, keep it wrapped in the swab.

Use alc swabbed scotch tape to cover inoculate hole.

Might help? Might not.
But, it is a very sterile procedure for inoculation.




i followed a similar sterile procedure for my LC jars.. but some are saying that that much alcohol is killing my spores??? http://www.shroomery.org/forums/showflat.php/Cat/0/Number/4271176/an/0/page/0


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OfflineSnarp
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Registered: 01/28/05
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Re: Rye jars keep contaminating, whats wrong?! [Re: lemunhed]
    #4272740 - 06/08/05 06:55 PM (12 years, 5 months ago)

I've done like Agar with the alcohol swab, but without any sterile environment at all. No problems. I just heated the needle from the syringe to the tip with a lighter flame and then put the tip in the swab, over the injection site, while still very hot. Everything in the needle is killed by heat, and nothing can get to it through the alcohol. Injection was through a self-sealing silicone bead over a small hole. There is no exposure of the inside to the outside except by the sterilized needle, so you can do it in a sewer without worry.

Um, don't set yourself on fire and don't dream of doing alcohol + flame in a closed space like a glove box. Fumes, whoof, owie.

I doubt it's endospores. You can PC a jar and it keeps for many months.


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OfflineIGnosticAbhorI
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Registered: 11/06/04
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Re: Rye jars keep contaminating, whats wrong?! [Re: Mych]
    #4272774 - 06/08/05 07:04 PM (12 years, 5 months ago)

Need to let those endospores germ.

Soak for 24 atleast.

-Gnostic


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Offlinelemunhed
The hustler'shustler

Registered: 05/12/03
Posts: 775
Loc: Ur moms house
Last seen: 12 years, 3 months
Re: Rye jars keep contaminating, whats wrong?! [Re: IGnosticAbhorI]
    #4273638 - 06/08/05 11:00 PM (12 years, 5 months ago)

I think its probably a combination of the rye being hard to sterilize and the method I cook it with. My bet is that not all the endospores are germinating. I'll try boiling. Agar thank you for your ultra sterile procedure, that's similar to what I do but without pushing the needle through an alcohol swab, I figure it might not be good for the spores... but thats just a feeling, no proof. As far as tyvek goes, I love the stuff but i think i'll switch to plastic lids with it, as it looks like some of the tyvek was getting sliced by the metal bands when screwing them on.


--------------------
You're pompeius, aren't you? I'll make a pompeian of you unless you hold your toungue!

-Tiberius Claudius Nero


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Amazon Shop: ½ Pint Jars, Agar, Rye Grain, Wild Bird Seed

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