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OfflineYossarian22
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Registered: 09/12/07
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contam problems
    #7955651 - 01/30/08 01:42 PM (16 years, 2 days ago)

I'm incubating at room temperature currently several quart jars filled with rye and inoculated with spores(PESA). I followed the rye grain teks here. I prepared the rye by rinsing it until it left the water clean, then let it stew in warm water overnight, then boiled and placed in jars. Since my PC can only fit two jars at a time, I put the prepared and closed(with the lid, no tyvek until I'm ready to put them in the PC) jars in the fridge until I could PC them. I have an old PC that doesn't rock at optimal pressure. I PCed them for 3 hours, let cool overnight, then inoculated. However, since the entire batch took 2 days, I kept some of the earlier sterilized specimens in the fridge for a day. Everything was going great until I noticed some contamination in one specimen. I removed it from the rest, and dumped it yesterday(outside, in the building's dumpster. It smelled very sour and garbage-like. I know it's usually a bad idea to smell contams but I did the wafting thing they teach you in high school science and it wasn't in an enclosed space, so I hope it's not too serious).

Anyway, it seems like the same kind of problem's appearing on a couple of my other jars. It always seems to start on the very bottom(even though I could've sworn I was golden with water content). Keep in mind that in most of these jars the spots are very small, especially compared to the mycelium, which has colonized about 1/3 of the jar so far. I've included some pics of one of the jars.

I'm planning on segregating the affected jars, but I really hope I can save these ones. It's fucking freezing outside, so planting them's out of the question. I'm guessing it's bacillus. Is there any chance of recovery especially if the mycelium's so much bigger and presumably stronger? Would it help to shake the grains up so the mycelium can begin fighting? Can you think of any steps in my procedure which might have caused this? Also, one of my friends studies mycology and works in a lab- I know this may sound like a long shot, but would there be any anti-bacterial agent someone in a lab might have easy access to that could help? Fucking bacteria.

So here are the pictures(You may have to zoom in to notice them):






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Offlinexeallos
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Re: contam problems [Re: Yossarian22]
    #7955690 - 01/30/08 01:50 PM (16 years, 2 days ago)

Quote:

Is there any chance of recovery especially if the mycelium's so much bigger and presumably stronger? Would it help to shake the grains up so the mycelium can begin fighting?




Not in my experience, no. A heavy bacillus infection is very slimy and spreads around a lot more readily than the mycelium will be able to recolonize after shaking. Once I see bac I consider the jar stalled and a lost cause, unless it is 99% and ready to be spawned and there is one tiny wet spot.


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OfflineYossarian22
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Re: contam problems [Re: xeallos]
    #7955764 - 01/30/08 02:03 PM (16 years, 2 days ago)

Fuck. There goes all but one of my jars. I put the contaminated jars across the room in a paper bag; would that be a problem? I'm holding out hope despite the odds, but if it would decrease the prospects of my healthy jar, I'll just toss 'em.


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Invisibleim_on_a_boat
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Registered: 04/06/06
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Re: contam problems [Re: Yossarian22]
    #7955779 - 01/30/08 02:06 PM (16 years, 2 days ago)

i haven't had luck with rye yet either..

wbs on the other hand.. :laugh:


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OfflineYossarian22
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Re: contam problems [Re: im_on_a_boat]
    #7956109 - 01/30/08 03:24 PM (16 years, 2 days ago)

Yeah, and I chose rye partially because it was supposed to be more contam-proof. Damn.

Another desperate question: since the infection is limited to the bottom of the jars, can I salvage the mycelium at the top and use that to spawn to bulk or is the entire jar crawling with the little bastards?


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Offlinexeallos
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Re: contam problems [Re: Yossarian22]
    #7956471 - 01/30/08 04:44 PM (16 years, 2 days ago)

Quote:

can I salvage the mycelium at the top and use that to spawn to bulk or is the entire jar crawling with the little bastards?




I have experimented with this and produced nothing but failure after failure. I have utilized the colonized sections of 15 bacillus infected qt jars in one monotub and the result is a waste of substrate materials, prep and layering time.


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OfflineRogerRabbitM
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Re: contam problems [Re: xeallos]
    #7957309 - 01/30/08 07:12 PM (16 years, 2 days ago)

After boiling the grains, you must put them in a filtered jar and pressure cook right away. Bacterial endospores, once germinated, form new endospores within hours. By putting them in the refrigerator for a day or two, you got a whole new bacterial bloom, some of which appear to have survived the pressure cooker.

Toss them and start over. Make sure your filter material is the right stuff too.
RR


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OfflineYossarian22
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Re: contam problems [Re: RogerRabbit]
    #7964855 - 02/01/08 01:01 PM (16 years, 13 hours ago)

All right. Thanks everyone for their help. As I mentioned, a friend works in a mycology lab so we might have a chance of using some antibiotics to fight them off, if only for the sake of science. Would anyone have any suggestions for how to apply them? I imagine too much liquid solution would throw off the moisture content; would a small amount shaken heavily do anything? I have one jar that's remaining. I didn't see any of the telltale white stuff around the grains, but I did notice that the grains on the bottom were becoming a little more shiny like in the other jars before the infection broke out. I decided to shake the jar up since maybe the water had accumulated at the bottom and was making that part a breeding ground for the bacteria. Mycelia seems strong and after the shake are appearing all over the jar. I assume that if the mycelium manages to outrace the bacillus that it shouldn't cause any further problems?

Also, my other batch, using a different syringe, has green mold in every jar right where the spore solution fell at the top. I used a Tyvek covering and micropore tape(on both sides of the injection holes, and then I applied an alcohol-swabbed piece of micropore tape following inoculation). Would this mean the syringe was at fault? I got it from a very trusted Shroomery sponsor, but I find it weird that none of the jars from the other syringe had that problem.


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