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deathlyego
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CVG, Pasteurization, and Trichoderma
#20653228 - 10/03/14 12:35 PM (9 years, 3 months ago) |
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Hello,
The situation: I've never done monotubs before. I have two in the pipeline. The first I might have fucked by putting on the casing CVG right after pasteurizing when it was still 85-90 degrees or so. I figured pouring it would get it down to 84F which is good temp for growing mycelium, so I thought it wouldn't be a problem. I got trich though that started growing on top of the casing layer, and then worked its way down. I took a 1/6th sized chunk out of the monotub and salted the wounds, then changed tools and scraped off the casing layer, it seems to be fine.
In the mean time, I have 2 more jars of CVG from the same pasteurization batch. No trich apparent in the jars, but I'm really nervous about using it to case after last contamination, so I have a couple questions:
1. How resistant is properly pasteurized CVG to Trich? Is my first batch getting trich probably caused by not cooling it enough so that the beneficial bacteria didn't have time to grow? Or can it get Trich regardless?
2. Should I re-pasteurize the same CVG before using it on second monotub casing? Does pasteurization even work against trich?
3. I'd rather not leave the PE uncased as I read it causes mutant blobs, however is it worth the risk if I have a known casing / trich problem? I assume there are spores all over the place, but perhaps it isn't a problem depending on question #1.
Any advice would be helpful... I really bummed me out to see first monotub fall to trich, really want the second to go better but just clueless whether it was simply not letting the CVG cool enough or whether I should just consider casing too high risk to do again.
Thanks
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Pastywhyte
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Re: CVG, Pasteurization, and Trichoderma [Re: deathlyego]
#20653439 - 10/03/14 01:28 PM (9 years, 3 months ago) |
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Coir is contam resistent regardless of how you prep it. You can sterilize it, bucket it, or pasteurize it, won't really matter.
If your tub triched out before first flush your spawn was dirty. Work on making clean spawn.
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deathlyego
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Re: CVG, Pasteurization, and Trichoderma [Re: Pastywhyte]
#20653786 - 10/03/14 03:19 PM (9 years, 3 months ago) |
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Alright, so I smelled all my jars and they smelled completely fine, all completely white healthy mycelium. I then spawned them, and they fully colonized, again without any signs of contamination. Because it is PE, I let the tub consolidate for 5 days... again no signs of contamination.
After all this being completely free of contamination, I put on the recommended 1/2in of casing, and it contaminates almost right away from the TOP. It was very clearly not reaching all the way through the casing layer. It started in a small patch from the surface itself.
Given all this, how can you say it was definitely the spawn? I inoculated from agar and have taken further samples during g2g and all of them are completely clean.
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Psilicon
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Re: CVG, Pasteurization, and Trichoderma [Re: deathlyego] 1
#20653817 - 10/03/14 03:29 PM (9 years, 3 months ago) |
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Trichoderma is incredibly difficult to isolate from cubensis mycelium or vice versa, because of Trichoderma's parasitic nature.
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Pastywhyte
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Re: CVG, Pasteurization, and Trichoderma [Re: Psilicon]
#20653946 - 10/03/14 03:59 PM (9 years, 3 months ago) |
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Quote:
van der griegen said: Trichoderma is incredibly difficult to isolate from cubensis mycelium or vice versa, because of Trichoderma's parasitic nature.
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elasticaltiger
Like Tigers in Coitus




Registered: 06/24/13
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Re: CVG, Pasteurization, and Trichoderma [Re: Pastywhyte]
#20653979 - 10/03/14 04:08 PM (9 years, 3 months ago) |
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He said the contamination was only on the casing layer though. Why would spawn contaminate a layer that it hasn't even made contact with on top? Unless of course there was trich mycelium on top already and he rubbed the casing layer over it when he was spreading it.
-------------------- First time growing cakes? DON'T make a Shotgun Fruiting Chamber The Shmuvbox. - The Old TC's Like it Afraid to Start Growing From Your Own Prints? Drop it Like a Tiger! No Pouring. No Syringes. No Cutting. No flaming. No Contamination. No Bullshit. "The best thing to do while your waiting is to start more stuff. I usually got so much happening that I have tossed projects simply because I didn't have time for them. -Pastywhite QFT Pastywhite's Easy Agar Tek (PastyPlates) Tiger Drop Video Demos By munchauzen Van Gogh would’ve sold more than one painting if he’d put tigers in them.―Bill Watterson EZEKIEL 23:20
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Pastywhyte
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Re: CVG, Pasteurization, and Trichoderma [Re: elasticaltiger]
#20654045 - 10/03/14 04:19 PM (9 years, 3 months ago) |
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Plain coir will not contam. I have tried to make it. The trich was either meshed in with his myc or his spawn was contamed with something else that allowed the trich to move in. Usually bacteria will do it for the second one. Any sub I ever suspect of being bacterial is fruited without casing because if you case a bacterial sub, the weak or poorly colonized grains having a microclimate provided will often trich out very quickly. It can appear that the coir contamed in such an instance but I assure you it didn't.
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bodhisatta 
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Re: CVG, Pasteurization, and Trichoderma [Re: elasticaltiger]
#20654065 - 10/03/14 04:22 PM (9 years, 3 months ago) |
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what is your casing layer? how was that prepared? also you can't cut trich out and you can't salt it out either. you're not salvaging a trich grow
like pasty said it was likely bacterial which if it's not a complete failure can be hard to detect. you'll get full colonization but usually some contam will set in before/during/ or right after the first flush then.
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deathlyego
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Re: CVG, Pasteurization, and Trichoderma [Re: elasticaltiger]
#20654093 - 10/03/14 04:26 PM (9 years, 3 months ago) |
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van der griegen: Are you saying Trichoderma doesn't show up on agar? because I've seen trich on agar, and none of this PE has it. I took multiple random samples from the same batch in question as I was spawning (pre-dunk) and the agar is completely clean.
Pasty... if I have two tubs that colonized the same CVG over several days without contamination, it consolidates for 5 days without contamination, and then after casing it contams.... how is that explained? I dug out the casing, the trich hadn't even made it through the casing layer, and after removal + salting the area, its been a week into fruiting with no more trich.
I realize this might be an exception to the norm, however I think I have a lot of solid evidence including the clean agar to suggest my spawn is fine.
I also realize CVG is known to be resistant, however I'm wondering if the factor was skipping out on a lengthy cool to allow beneficial bacteria to grow, and instead exposing it to the spawn while it was still above 85F, which my understanding is helpful for trich growth.
Right now, my second monotub is from the same PE monoculture and it has been consolidating for 5 days and looks just perfect. I'm just wondering if its worth the risk to case, or whether the week the CVG has been left to sit after pasteurizing has made it resistant enough to trich to use it.
Also, **regardless** of the source of the contam, can anyone answer my questions regarding how important the cooling step is for pasteurization, whether pasteurization above 140F for 90 minutes kills trich, and how long it takes for the bacteria to populate enough to make it trich resistent?
I'm mostly worried about contaminating my perfect second tub with the casing layer. I'd rather risk blobs than contaminate it at this point.
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elasticaltiger
Like Tigers in Coitus




Registered: 06/24/13
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Re: CVG, Pasteurization, and Trichoderma [Re: deathlyego]
#20654131 - 10/03/14 04:30 PM (9 years, 3 months ago) |
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Yes pasteurization above 140 for 90 minutes will kill trich.
I'm not sure I understand what you're unsure of in the cooling process?
Stuff needs to cool down. I've spawned in 90 degree CVG before (don't recommend it though, just saying)
Never cased with warm coir though.
-------------------- First time growing cakes? DON'T make a Shotgun Fruiting Chamber The Shmuvbox. - The Old TC's Like it Afraid to Start Growing From Your Own Prints? Drop it Like a Tiger! No Pouring. No Syringes. No Cutting. No flaming. No Contamination. No Bullshit. "The best thing to do while your waiting is to start more stuff. I usually got so much happening that I have tossed projects simply because I didn't have time for them. -Pastywhite QFT Pastywhite's Easy Agar Tek (PastyPlates) Tiger Drop Video Demos By munchauzen Van Gogh would’ve sold more than one painting if he’d put tigers in them.―Bill Watterson EZEKIEL 23:20
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deathlyego
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Re: CVG, Pasteurization, and Trichoderma [Re: deathlyego]
#20654139 - 10/03/14 04:32 PM (9 years, 3 months ago) |
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bodhisatta: It is 1 coir brick, 2 quart verm, 1 handful of gypsum to field capacity. It is the same as my spawn, which colonized perfectly.
I scooped it into jars, pasteurized with thermometer for 60 minutes above 140F, peaking at 162F. The only shortcut I made was that I only waited for the jars to cool to 85F-90F before dumping them into a bowl and then casing. My worry is that the beneficial bact didn't have time to grow strong enough to fight of trich that I picked up from the air.
I realize the advice on trich grow, and I also realize there are reports of it working for people. I can say this, I scraped off the casing layer, and since that the rest of the remnants of the casing layer have fully colonized cleanly and the rest of the tub looks fine. There is no uncolonized area of the tub except for the salted ring around the hole I cut out. no growth trich or otherwise in that salt ring. Rest of tub is fine. Again, I should mention I caught the trich before it sporulated, and before it reached the spawn. It had not penetrated the casing layer. I cut out a hole in the sub just to be extra safe, however I don't believe it got through to the spawn.
I'll keep my eyes out for bacterial contam, but really all the spawn looks completely clean, no extra moisture or metabolites, and the agar coming out of the jars during spawning looks completely clear as well. I understand the skepticism, but I have a very strong scientific method and lot of evidence to suggest the trich started on the casing layer and the spawn is fine.
The second tub is a g2g from the first batch, should I just fruit it un-cased to show that it is fine? I'd rather go for some non-blob spore producing from casing, but I'll leave it uncased if you want more evidence that the spawn is clean.
Edited by deathlyego (10/03/14 04:35 PM)
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bodhisatta 
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Registered: 04/30/13
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Re: CVG, Pasteurization, and Trichoderma [Re: elasticaltiger]
#20654148 - 10/03/14 04:34 PM (9 years, 3 months ago) |
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you don't want a lengthy cool down you want it to cool down at room temperature. some people call that lengthy some call it fast. either way you don't want the jars to cool down in the hot water bath that's too slow and can even kill the thermophiles. but you don't want to put them in cool water either, you want to take them out and let them cool in room temperature air, should take a few hours to get back to room temp in the center of your jar.
trich shows up on agar but can grow along with cubensis mycelium leading even some of the veteran hands to accidentally spawn trich because it's acting almost like a "hidden contam"
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deathlyego
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Re: CVG, Pasteurization, and Trichoderma [Re: bodhisatta]
#20654163 - 10/03/14 04:37 PM (9 years, 3 months ago) |
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Yeah, I had just let it cool down room temp on the counter until it was 85F-90F. I was following Franks Proper Pasteurization 3.0 which calls for 12 hour cool down. I wasn't sure if that was just to be safe that it was room temp, or whether the extra 6-8 hours was to ensure proper bact growth.
So... whats the verdict on my second clean monotub. Case with the same CVG (that has no sign of any contam in the jars)? Leave the PE uncased?
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bodhisatta 
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Loc: Milky way
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Re: CVG, Pasteurization, and Trichoderma [Re: deathlyego]
#20654173 - 10/03/14 04:39 PM (9 years, 3 months ago) |
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Quote:
deathlyego said: bodhisatta: It is 1 coir brick, 2 quart verm, 1 handful of gypsum to field capacity. It is the same as my spawnsubstrate, which colonized perfectly.
I scooped it into jars, pasteurized with thermometer for 60 minutes above 140F, peaking at 162F. The only shortcut I made was that I only waited for the jars to cool to 85F-90F before dumping them into a bowl and then casing. My worry is that the beneficial bact didn't have time to grow strong enough to fight of trich that I picked up from the air. doesn't matter anyway, you can skip pasteurization all together with coir verm gypsum and it won't trich out unless you have trich in your spawn, a lot of us have tried, we have even just hydrated CVG with room temp tap water and let it sit out for days and no mold grows on it. people use coir in reptile terrariums just hydrated no pasteurization and the coir doesn't mold for months. trich simply won't spontaneously germinate on coir, it hitched a ride somewhere.
I realize the advice on trich grow, and I also realize there are reports of it working for people. I can say this, I scraped off the casing layer, and since that the rest of the remnants of the casing layer have fully colonized cleanly and the rest of the tub looks fine. There is no uncolonized area of the tub except for the salted ring around the hole I cut out. no growth trich or otherwise in that salt ring. Rest of tub is fine.
I'll keep my eyes out for bacterial contam, but really all the spawn looks completely clean, no extra moisture or metabolites, and the agar coming out of the jars during spawning looks completely clear as well. I understand the skepticism, but I have a very strong scientific method and lot of evidence to suggest the trich started on the casing layer and the spawn is fine. I've had bacterial grows that the spawn looked clean, smelled clean, and had no metabs but was bacterial, sometimes it's just really hard to tell when there's no indications such as thick mycelium or smell, metabs etc...
The second tub is a g2g from the first batch, should I just fruit it un-cased to show that it is fine?try it you don't get blobs just from not having a casing plenty of people grow PE without casings and without blobs I suspect it has more to do with consolidation I'd rather go for some non-blob spore producing from casing, but I'll leave it uncased if you want more evidence that the spawn is clean.
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Pastywhyte
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Re: CVG, Pasteurization, and Trichoderma [Re: bodhisatta]
#20654185 - 10/03/14 04:41 PM (9 years, 3 months ago) |
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Coir can be sterilized. It doesn't have any beneficial bacteria to survive and spread. We simply need to cook it. If a grain on top was weakly colonized due to an endospore trich will germinate on it and move upward. I believe that endospores are a lot more common than most people realize and that many of us would consider such spawn to be clean.
Your consolidation times actually reinforce this. Endospores can germinate on already colonized grains.
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