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InvisibleZestyMycelium
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Stalled Jars...need advice!
    #6177223 - 10/17/06 12:51 AM (17 years, 4 months ago)

Ive progressed pretty well with the traditional PF-Tek/BRF cakes. I figured Id give whole grains a try and prepared some whole grain brown rice for inoculation using 1/2pt and 1 pt jars. At first colonization was ridiculously fast - the 1/2pt jars were done in a little less than 2 weeks and the pint jars were a good halfway through.

Then, everything stopped! About 3/4 way through, there was just no more mycelial growth. I tried all the methods I knew of to kick start things - shaking, exposing to dim light, etc. The Equadorians actually took to this and finished colonizing. My Tazmanians and South Americans are still barely over the 3/4 mark, now 6 weeks after inoculation. I probably didnt get the moisture content quite right or packed them a little too tight, etc.

I have enough mycelium to make some nice casings, but I used up the last of my prints for a couple of the strains and want to make sure I dont lose them in the event that fruiting is unsuccessful.

What should I do? Use the mycelium I have to spawn some bulk substrate? Would WBS do the trick?

Ive thought about creating some mycelium syringes as well, to further insure that I keep my strain collection well stocked:)

Thanks for any advice!!
ZM


Edited by ZestyMycelium (10/17/06 12:51 AM)


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OfflineBallsDeep
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Re: Stalled Jars...need advice! [Re: ZestyMycelium]
    #6177235 - 10/17/06 12:58 AM (17 years, 4 months ago)

I dont know about your stalled jars but you could try to make some Liquid Cultures. I only had 3 cc left and i made up 3 jars, all are looking great so far.



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InvisibleZestyMycelium
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Re: Stalled Jars...need advice! [Re: BallsDeep]
    #6177272 - 10/17/06 01:14 AM (17 years, 4 months ago)

Thanks for the tip, ballsdeep. Im looking at this tek:

http://www.shroomery.org/9145/Liquid-Culture-Basics

This calls for spores to be added to the liquid culture - could I do the same thing with a bit of colonized mycelium, working in a glovebox?

I see I can add H2O2 to the mix, perhaps I can even inject a syringe of liquid culture into my colonized jar, and then suck it back up again?

ZM


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OfflineBallsDeep
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Re: Stalled Jars...need advice! [Re: ZestyMycelium]
    #6177278 - 10/17/06 01:19 AM (17 years, 4 months ago)

yes those are both plausable methods. I have never done it though so I cant help you out there. I would suppose a light h2o2 mix would improve your odds or atleast add some safety.

Good luck and make sure to keep it sterile!


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InvisibleZestyMycelium
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Re: Stalled Jars...need advice! [Re: BallsDeep]
    #6177292 - 10/17/06 01:29 AM (17 years, 4 months ago)

Nice - this sounds like a plan then! I'll allow me to case what I have now and prepare some liquid culture for further experimentation:)


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InvisibleToTheExtreme, LOL
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Re: Stalled Jars...need advice! [Re: ZestyMycelium]
    #6177330 - 10/17/06 02:00 AM (17 years, 4 months ago)

I would boil some distilled water and shoot a cc or 2 into the cake with your spore syringe. I've hears this works. also check out my thread about stalled rye jars. There was some good info given there

http://www.shroomery.org/forums/showflat.php/Cat/0/Number/6176478/an/0/page/0


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OfflineBallsDeep
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Re: Stalled Jars...need advice! [Re: ToTheExtreme, LOL]
    #6177354 - 10/17/06 02:25 AM (17 years, 4 months ago)

Personally if you were going to go that route I would fill a syringe with distilled or R/O water, PC it, and then inject that.

Thats just me though!


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Offlinehyphae
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Re: Stalled Jars...need advice! [Re: ZestyMycelium]
    #6177498 - 10/17/06 05:23 AM (17 years, 4 months ago)

Do you have tape over the holes?


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InvisibleZestyMycelium
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Re: Stalled Jars...need advice! [Re: hyphae]
    #6177554 - 10/17/06 06:21 AM (17 years, 4 months ago)

I had the hole covered with electrical tape but removed it after I say the mycelium take hold.


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OfflineRogerRabbitM
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Re: Stalled Jars...need advice! [Re: ZestyMycelium]
    #6177726 - 10/17/06 08:06 AM (17 years, 4 months ago)

If something has caused the mycelium to stall in the jars, don't try to make liquid culture out of it.

If you have holes open for gas exchange and no visible water standing in the bottom of the jars, the biggest reason for stalling is bacteria. You can tell that by smelling through the holes in the lid. If it stinks, toss it out. What sort of filter are you using?
RR


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OfflineJorsher
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Re: Stalled Jars...need advice! [Re: RogerRabbit]
    #6177763 - 10/17/06 08:28 AM (17 years, 4 months ago)

I know it's been answered but I figured some extra comments would help.

I wanted to make an LC with the mexicube strain but didn't have spores, only myceleum. I sucked up alcohol with the syringe, then squeezed it out and let dry, then heated the needle until it was red hot, wiped with alcohol, sucked up some sterilized lc, burned the needle and wiped again, injected into a jar with some force to rip up some myceleum, sucked up what I could, sterilized the needle again, and injected into the lc. It worked great and so far there aren't contaminates, although I would suggest pc the syringe.

Good luck!


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InvisibleZestyMycelium
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Re: Stalled Jars...need advice! [Re: Jorsher]
    #6180843 - 10/17/06 10:56 PM (17 years, 4 months ago)

Quote:


If something has caused the mycelium to stall in the jars, don't try to make liquid culture out of it.

If you have holes open for gas exchange and no visible water standing in the bottom of the jars, the biggest reason for stalling is bacteria. You can tell that by smelling through the holes in the lid. If it stinks, toss it out. What sort of filter are you using?
RR





Standard PFTek Vermiculite filter...I dont think its bacteria. Ive dealt with that before, and it can really build up a stink in the incubation chamber after a couple of days. It almost seems as if the rice might be packed too tight in some spots? I know it wont break free even with vigourous shaking.

Ever since I started using a Pressure Cooker to sterilize and a glove box to inoculate, I really havent had a problem with contams. Even had an un-inoculated jar sit at room temps for nearly 2 1/2 months and when I cracked it open it still smelled fresh...

Not going to rule it out though!! I'll start a few more jars with BRF just cause it's my faithful standby.


Edited by ZestyMycelium (10/17/06 11:11 PM)


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InvisibleZestyMycelium
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Re: Stalled Jars...need advice! [Re: ZestyMycelium]
    #6263975 - 11/09/06 12:30 AM (17 years, 3 months ago)

Just an update here folks...

The stalled pint jars were due to lack of gas exchange.  When I made the jars, I covered the holes in the metal lids with electrical tape before sterilzing in the pressure cooker.  I'd always used masking tape before - and didnt realize the electrical tape was acting like a self healing injection site, obstructing airflow!

So I uncovered all the holes and the mycelium took off like a rocket -- and a week later all the uncolonized brown rice had turned slimy-nasty with bacteria.  This is the first time Id tried a whole grain tek - I made up a substrate of cooked whole grain brown rice with some dry ground flaxseed mixed in after it cooled to absorb and excess water (that worked really well actually...) The mycelium seemed to love the substrate - but I didnt realize that it doesnt stay 'caked' like BRF/verm cakes do.  The vermiculite filter I used ended up flowing down the sides of the jar - exposing the substrate to all manner of nasties once the holes were uncovered.

It seems like I need to fabricate some new lids using polyfill/tyvek if Im going to use whole grain rice with pint jars.  Half pint jars colonize extreemly fast with this substrate - like 10 days or so. Even with the holes left taped up, it seems like there's enough air to allow for full colonization with the half pint jars.  I had one that I inoculated 7/15, and just took it out and cased it a moment ago - it was completely fresh after removing from the jar.

Moral of the story - With whole grain BR, use nice filter-style lids with pint jars. Leave the half pints taped up.  I should have known better :smile:

ZM


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InvisibleBlutjager
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Re: Stalled Jars...need advice! [Re: ZestyMycelium]
    #6264632 - 11/09/06 09:02 AM (17 years, 3 months ago)

Tyvek is a million times better than polyfill


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