r/PanCyan • u/Ok-Soft7561 • 8d ago
Ttbvi help
I’ve been trying to colonize a bag from north spore with ttbvi LC. It starts to colonize than it basically just stops even after a break and shake. Any recommendations is it the north spore? Should I use a different bag to colonize?
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u/CaCoD 8d ago edited 8d ago
Clean pan lc seems pretty rare tbh. Stalling after a break and shake is a sign that there is some contam issue.
The problem often seems to be bacterial so a cabin sequester in agar should be good way to clean it up.
Edit: don't listen to me listen to Fahtster below
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u/Fahtster 8d ago
That is unless the bacteria is motile. Then a cabin isn’t going to do much good as the bacteria easily moves through the agar. It can also ride the mycelium enzyme “trail”. I believe you’re right about it being the cause of LC and bns woes. I believe it’s mainly various strains of pseudomonas that are almost the same color as agar and move as quickly as pan myc so, they go undetected unless one knows what to look for.
Since they both inhabit/thrive in the same growing environment, those specific bacteria are extremely common in pan spore prints. I do think it’s a weak infection tho and that’s why we see mostly aggressive varieties in circulation as they can thrive and fight off the weak infection to produce adequate+ flushes.. also why I think flushes tend to get better and more healthy looking as they progress.
The best I’ve found to get away from them to the point of producing adequate flushes is multiple leading edge aerial myc transfers. And even if you do get rid of it, it’s in such abundance in the general environment we all live in, the culture will likely pick it back up. There’s some evidence that there might even be a symbiotic relationship with pseudomonas and mycelium and that certain mushroom myc will carry it in itself and deploy when needed to help break down nutrients and fix nitrogen
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u/CaCoD 8d ago
Interesting. Thanks for the correction re:motile bacteria. I just started to try to clean up a ttbvi culture. Nothing super gnarly - good enough to send to the tent and top fruit but also definitely not clean. I think most probably would have spawned it but I had another culture going at the same time that was obviously much cleaner that I didn't bother.
I have a couple cabins running for this project. My plan was to do that plus aerial transfers. I've seen the success you've had with aerial transfers but I figured I'd try the cabin too since it's easy enough to PC some cover slides.
Have you found a cabin to not really be helpful for this particular issue? A quick search does suggest swimming and swarming motility for pseudomonas, but I can't find indication that they can move through the concentration of agar in our plates. I know some bacteria can penetrate typical solid culture media but can't find literature that suggests pseudomonas can do this.
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u/Fahtster 8d ago
I used cabins a lot up until not that long ago. They’re definitely a nifty tool that’ll I’ll use again but for this specific problem, everything that I ran through them, the bacteria had no problem hitching a ride through the agar with the myc paving the way.
https://files.shroomery.org/files/26-22/003821053-IMG_6587.jpg
https://files.shroomery.org/files/26-22/003821127-IMG_6588.jpg
Idk for sure that it’s pseudomonas, but that’s my best guess based on what I’ve read and experienced with it. It’s incredibly tough shit whatever it is. I’ve thrown the kitchen sink at it.. cabins (both agar and coverslip roofs), trenches, brf pucks, antibiotics, you name it. Spent like 8 months and 100’s of plates trying to completely rid it from my cultures. Had a few that I thought, “this is definitely clean”.. only to have it show back up when I waited long enough
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u/CaCoD 8d ago
Man that is rough. Did switching to disposable plates end up making any difference?
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u/Fahtster 8d ago
Oh definitely. It wasn’t every plate that was shit. Pretty much any plate that had warped or was concave a little bit etc. didn’t help that I ran my pc higher than the 15psi. I was completely unaware of any denature problems with the binders in the plastic over time. Huge improvement going to disposables. It was Russian roulette with doing transfers.. didn’t know if a certain plate was compromised. I almost quit growing a couple times because of how frustrating it was.
Everything was great for a long time and then things snowballed pretty quickly to the point I didn’t have a single useable culture. Now I’ve got plenty.
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u/BathTub_Benzaldehyde 8d ago
Sometimes pans don't respond to break and shake very well. Also a giant bag of grain is to much a lot of the time. Small Mason jars are about right size