jee3apha said:

jee3apha said:

So I tried to transfer from these two plates (the middle denser spot of the latter), which in the meantime, they too started developing the halo as well:

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Some updates about the transfers.

[...]

Another was some mild but seemingly clean mycelium, while the last one was this very strong, white and tomentose mycelium:



This is the first time I see something like this, but looks legit, what do you guys think? I plan to transfer from this shortly.

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So I did, I transferred to some other plated which basically grown similarly, just a bit less strong, and then today I used them to inoculate some jars. Will keep you posted...

I kept the donor plate of those sent to grain just to see how it went (notice the holes from previous transfers), I also scratched the surface to better appreciate the smell, and it was good mushroom-like, also you can see that it started to bluify a little, which is a good thing, I guess, at least IME.

jee3apha said:

So I did, I transferred to some other plated which basically grown similarly, just a bit less strong, and then today I used them to inoculate some jars. Will keep you posted...

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And here's how the jars look like, will wait a bit more then shake. IMHO they look clean and nice, we'll see... do they look legit in your opinion? Any red flag?

jee3apha said:

jee3apha said:

So I did, I transferred to some other plated which basically grown similarly, just a bit less strong, and then today I used them to inoculate some jars. Will keep you posted...

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And here's how the jars look like, will wait a bit more then shake. IMHO they look clean and nice, we'll see... do they look legit in your opinion? Any red flag?

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So I started a monotub from these jars, and after a month there a are still no pins, and at this point I strongly doubt it will ever grow anything. The mycelium looks white and heavily tomentose, similar to the above jar.

Here's a picture after 5 days:



But eventually some yellow/brown droplets started to appears in a handful of spots, most of the mycelium is stile white and clean. I've never saw anything like this, I immediately thought this is what people call "mushroom piss", or maybe it's just some kind of contamination:



In any case, then I really wonder what's this if it's not cubensis mycelium, can't be some random mold I guess, after all it came from a print. I mean, one thing is some clearly contaminated mycelium, another is this thing. Also it looks awfully similar to my earlier monotub (different spore source), which likewise yielded no good fruits:



Which in turn, is quite different from a good one I had, by looking at the substrate surface:

langten69 said:

You can take a fork, and rake the surface to till it, and then mist the surface to make it have "perfect surface conditions" (ie. Countless water droplets)

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jee3apha said:

So I started over again, inoculating some more plates with the same print, using a sterile swab, and here's what I got:



I then transferred in some more plates, and I'm pretty sure I'll end up with the same halo as before, this time though I'll try to fruit it, if I can. Will post updates...

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Quote:

Some strains produce random monokaryotic filaments that eventually populate with the complementary nucleus. This is noticeable as a "halo" that is decidedly not bacterial (though, bacterial halo is definitely a thing). Certain environmental and chemical exposures can increase the occurrence.
https://www.shroomery.org/forums/showflat.php/Number/27927633#27927633

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Rasmodius said:

I bumped into this in one of my favorite threads over the weekend, and thought of you:

Quote:

Some strains produce random monokaryotic filaments that eventually populate with the complementary nucleus. This is noticeable as a "halo" that is decidedly not bacterial (though, bacterial halo is definitely a thing). Certain environmental and chemical exposures can increase the occurrence.
https://www.shroomery.org/forums/showflat.php/Number/27927633#27927633

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That thread is very dense with a lot of crazy shit that isn't relevant here, but this was interesting because it sounds like we've both seen it on our plates: a distinct halo that (sometimes) eventually fleshes out into stronger and more obviously correct myc.

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Will look into that, thanks!

Quote:

Your monotub originally looked very well colonized, before getting that spot of wet bubble. The wet bubble is localized to that one small area, and it's right on the wall, so I suspect it came in from water drippings off the walls. If it was from the plate and rode along on the spawn, you'd have seen it across the tub, and earlier. I have one semp plate with wet bubble and the center transfer threw droplets like that before the myc reached the walls. Certainly your older plates would have gone that way by now. What's more interesting is how fluffy everything else is in that pic, that seems wrong/unusual.

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I honestly don't think it came from the walls, that was not the only spot, there were like 5 others, yes near the border, but never always too close to it. I'm almost positive it came from the plate, still the vast majority of the surface was unaffected, so that wasn't (solely) a brown bubble issue. IMHO it came from below, from some kind of blob, I'm not sure.

Yes the monotub was very well colonized, but way too fluffy, and IME that kind of fluffiness always led to trouble. If I compare that surface to the one of my most successful monotub I notice that the latter is not just a fluffy bed of mycelium. Here's what I mean:

Quote:

I gotta ask, just out of curiosity: why this culture? It seems like you've fruited other tubs with success. Unless this is a very sentimental lineage or something, if it were me, I'd be looking for a different starting point and probably walk away from a spore source that was giving me hell like this.

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Eh, because after my latest successful monotub I couldn't replicate, you can take a look at my past threads if interested, but basically this: https://www.shroomery.org/forums/showflat.php/Number/29312028 (where you can also see the difference in terms of monotub surface: fluffy surface = badly dying fruits).

I kept using the original spore print I had, and some others that I did myself, and couldn't reach proper fruiting anymore, so I bought another print and this thread is documenting my struggle with that. So basically I have like: the original print, a couple others I created, this new one I bought. I guess I could try again with the older ones, but I mean, I tried already.

Despite what some members say, I'm pretty confident my sterile procedure is decent, every issue I had always came from the spore source in some way or another, meaning that pretty much never happen that contamination pops up in the middle of the agar for no reason, rather it's always tied to transfers of inoculation material.

Quote:

Your germination plates are very dense, which is expected if you're swabbing them directly. If you wanna keep going with this spore source, one next step could be serial dilution: tweezer a few spore-carrying swab strands into a small amount of sterile water, shake the everloving shit out of it, and streak a plate with that. There will be much less germinating material per unit of volume, which means that both spores and contams are farther apart on the plate, and can be transferred or avoided in isolation. If it's still too dense, use a sterile syringe to move a sample of that liquid into another volume of sterile water, diluting it further (that's the "serial" part), until you achieve a density that lets you get the distances you need.

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That's good to know, thanks for that!

Yes, indeed they are very dense, but I was experimenting with streaking, I'm still learning the proper technique so I'm playing around a bit, the above plates were expected to be that dense, hopefully won't happen in the next attempt. (I guess the mistake was to keep smearing with the side of the swab filled with spores, hence effectively not diluting the concentration at each segment, but rather keeping drawing a spore line again; sorry my English is lacking here, not sure if I made myself clear.)

DV8DUG said:

Any reason you don't take pix with cover off? Be nice to see the plate better.

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Rasmodius said:

I bumped into this in one of my favorite threads over the weekend, and thought of you:

Quote:

Some strains produce random monokaryotic filaments that eventually populate with the complementary nucleus. This is noticeable as a "halo" that is decidedly not bacterial (though, bacterial halo is definitely a thing). Certain environmental and chemical exposures can increase the occurrence.
https://www.shroomery.org/forums/showflat.php/Number/27927633#27927633

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That thread is very dense with a lot of crazy shit that isn't relevant here, but this was interesting because it sounds like we've both seen it on our plates: a distinct halo that (sometimes) eventually fleshes out into stronger and more obviously correct myc.

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About the halo, today I noticed that it appeared even after one of my previous attempts (another spore print) with Cabin-Sequestering (see the left side):



Not sure what to make of it, maybe it's somehow reassuring. Will read your thread...

Rasmodius said:

I wish I had more to offer here. I am not competent enough at myc ID in the scope to speak with the level of certainty I like to for advice posts.

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Quote:

But this definitely biases me towards "this kind of halo is normal myc, some varieties just do that". Going forward I won't consider this kind of thing a contam indicator unless another piece of evidence points toward it, I think.

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You're probably right, let me add that the above monotub was started with some mycelium that apparently didn't have that halo (I tossed those plates, and inoculated again IIRC), this time instead I will try to see, and possibly send it to fruiting.

But as I probably already said, it wasn't really the halo to worry me, rather is the fact that seemingly good mycelium, in presence of the halo appears to behave funnily, see for example this latest picture:



I mean, this sounds like a red flag to me.

DV8DUG said:

Any reason you don't take pix with cover off? Be nice to see the plate better.

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OK so since I tossed some of the plates, here's a possibly better picture of the uneven one. You can even see a small yellowish droplet (I'm pretty sure it wasn't so yellow before), which to me looks awfully similar to the brown bubbles in the above monotub.