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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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Tomatose myc on agar, safe to transfer to grain?
    #26913144 -

So, started again from spores, right now at 4th transfer, still no rhizo myc, have had problems with contamination before, so would like to be sure if they look okay to transfer to grain or are they off, is something maybe riding along? Transfers were always made from the transparent seeking part of mycelium, they look like little filaments. But as they grow they always turn out this fluffy.



Edited by otsilla (09/02/20 04:26 AM)

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Re: Tomatose myc on agar, safe to transfer to grain? [Re: otsilla]
    #26913186 -

Have you tried limiting your sugar / dextrose / nutritional source in your agar recipe? Sometimes mycchorizae does not go rhizo unless it is starved.

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Re: Tomatose myc on agar, safe to transfer to grain? [Re: otsilla]
    #26913205 -

I used 10 agar 10g potato flakes 7g dextrose, last time I got rhizo myc with the same recipe, idk, havent made another batch of agar yet, problem isn't so much in rhizo than to spot contaminants. I'm worried that something might be hitching a ride. Can't be sure before I move to grain? I mean its 4th generation and they look uniform and okay, + I always took transfer from the transparent looking edge, so should be fairly fine?

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Re: Tomatose myc on agar, safe to transfer to grain? [Re: otsilla]
    #26913254 -

Sure you can do.


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looking for a sclerotia producer print

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Re: Tomatose myc on agar, safe to transfer to grain? [Re: Martinsapin]
    #26913332 -

Tomentose is fine, but I don’t like those plates. Something just looks wrong to me.

This is the most tomentose culture I’ve ever had.


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LAGM2020

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Re: Tomatose myc on agar, safe to transfer to grain? [Re: otsilla]
    #26913337 -

weird, what could it be tho? they've always been this fluffy for me, they start out transparent, you can barely see againts light tiny filaments that seem to be myc, so at first i always transfered those so called transparent parts, 4 transfers in they look the same. also almost out of spores so i really need to clean this up somehow.

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Re: Tomatose myc on agar, safe to transfer to grain? [Re: otsilla]
    #26913340 -

Mold, or mold meshed in maybe. Or bacteria.

There’s some decent looking myc on the early plates, I’d grab some of that.

You could try hot pouring the plates you’ve got now that look funny. Just make more agar and pour a thin layer right on top of the myc. Hopefully it’ll grow through to the surface and leave whatever else is in there behind.


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LAGM2020

Edited by A.k.a (09/02/20 08:09 AM)

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Re: Tomatose myc on agar, safe to transfer to grain? [Re: otsilla]
    #26913350 -

from which plate and place exactly?

haven't heard about the hot pour, that looks promising tho, i might try that also, how thing of a layer should it be and how do i do transfer from there, just scrape?

Edited by otsilla (09/02/20 08:15 AM)

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Re: Tomatose myc on agar, safe to transfer to grain? [Re: otsilla]
    #26913369 -

The hot agar pour is a way. Just read that in Peter Mccoys book yesterday.
Also you could change you recipe, if youre eager to do a fifth transfer

All my LGT platesT2 are fluffy asf at the moment, and as Im using grain water agar for the first time wondering if it could be that.

Aka what makes you believe there might be mold or bacteria riding along? Apart of the fluffy look?

LGT on GWA




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looking for a sclerotia producer print

Edited by Martinsapin (09/02/20 08:47 AM)

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Re: Tomatose myc on agar, safe to transfer to grain? [Re: otsilla]
    #26913381 -

I might put one plate straight to grains, then do hot pour on some/rest of them to have some backup in case it fails.

Well not much more besides that they are fluffy asf, when searching fluffy/tomatose myc, there are really few images on this forum that are that fluffy. Also because my second attempt from agar to grain also failed because of bacteria. Although my sterile procedures have gotten way better as there were no bacterial colonies on my last batch of agar plates.

But id like to be 100% sure that it's clean before moving to next step just to not to have to start all over again and also because the lack of spores.

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Re: Tomatose myc on agar, safe to transfer to grain? [Re: otsilla]
    #26913390 -

Those plates seems to be bacterial to me.
I wouldn't inoculate them; they will be more prone to contamination.
You could either hot pour as suggested or use antibiotic agar. I've never had much success with the former though.
Chances are that If you inoculated from syringe, it would be that. Or maybe another source of bad hygiene, as maybe badly sterilizes scalpel, touching the plates with the gloves, and so on. There are infinite possibilities.

Edited by gabbk (09/02/20 09:09 AM)

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Re: Tomatose myc on agar, safe to transfer to grain? [Re: gabbk]
    #26913434 -

Dont we spot, most of the times, bacteria by the wet spots they produce and the way they inhibit myc progression on the plate.
Also they appear in odd areas of the plate usually and near the edge

Still not convinced its bacterial...


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looking for a sclerotia producer print

Edited by Martinsapin (09/02/20 09:27 AM)

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Re: Tomatose myc on agar, safe to transfer to grain? [Re: Martinsapin]
    #26913544 -

Just try spawning it... It will be more prone to contam, but if it doesn't contam (which is generally by the mean green Trichoderma) you most surely will get delayed performance, both speed growth and final size, resulting in a notoriously lower final yield compared to if it was a 100% healthy and clean spawn from the get go.

I struggled with the same issue also for a long time when I began growing, until I could visually tell. Some of these tubs birthed with the same looking bacterial agar took up to 21 days from birth to pin.
Had to search in my gallery; these agars looked were very similar to the ones shown by the op:


Bacteria can manifest in lots of ways, because there exists tons of different types. Some of them can form colored stains or goo, some others just wet spots as you indicate... And some of them just 'molest' the mycelium by invading them and not letting it grow normally, acting like a competitor. You could see the bacteria IN the mycelium under a microscope, but macroscopically you can see it like really really cottonish, like a very poorly structured mycelium, particularly with fuzzy edges. Just compare otsilla pictures, mine, and then watch that clean tormentose growth A.k.a shared.
Sometimes it can be tricky to spot the difference, specially when you begin to confront this issue.

Another tip that goes with this kind of bacteria is that sometimes it can form a halo underneath the mycelium leading it; also sometimes difficult to spot, but easier to recognize against a light, just like in the following picture:

That's bacteria overpowering the myc.


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Edited by gabbk (09/02/20 11:17 AM)

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Re: Tomatose myc on agar, safe to transfer to grain? [Re: otsilla]
    #26914810 -

So last night I hot poured 3 of them,left only one to put to grain. The fluffyness was quite tall so had to pour a lot to cover it fully, in the center it got only a thin coating tho. But it leaved quite a little space between the petri the cover and top, but should be sufficient. I guess we'll see in the following days.

Also gabbk pictures are on point. It started exactly like on the second picture, although I always made transfers before they got to that phase with swirling ridges from that really transparent edge.

I think that I have just dirty spores, hope that the hot pour tek works.

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Re: Tomatose myc on agar, safe to transfer to grain? [Re: Martinsapin]
    #26914839 -

Martinsapin said:
Aka what makes you believe there might be mold or bacteria riding along? Apart of the fluffy look?



I agree with others on the contam theory.  When there's bumps and blips and lumps in there it's often because the mycelium ran into something.  Unless you're agar is extra chunky.


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Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here.
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Have some medicinal mushrooms and want to get the most out of them?  Try this double extraction method.

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Re: Tomatose myc on agar, safe to transfer to grain? [Re: Forrester]
    #26915032 -

Great!

Now you gotta pay close attention to the visual aspect of the grain. Here it is another commentary I made on another user's post that seemed to me he was encountering bacterial contam also, originated/carried by the inoculum (agar) and it may help you!

gabbk said:
I'm pretty sure the spawn was bacterial. All my bets on it.

Last week I had a tub that also was 2 weeks into birth and wasn't giving pins in spite of being 100% colonized for a good week, until it gave up on me turning green (I had a pretty big Trich contam 3 weeks ago that blew up my whole grow room and didn't know until then, when that tub turnt green and had to think why it was, as I've never encountered green mold :sad::mad2:)
Maybe your tub won't turn green because your grow room maybe has little Trichoderma spores floating around, and if it's the case, chances are you will get at least some fruits; just when I began growing, my very first took 21 days from birth to pin. Worth to mention again that that clearly was allowed by the fact there wasn't the same quantity of Trich spores in my grow room like now, and the mycelium could grow. It really took a blue moon but gave something anyways.

By then I was also struggling with bacteria in the spawn but didn't know because I couldn't recognize the visual difference between the bacterial mycelium and the healthy mycelium. And I'm not talking about those black or colored nasty stuff that anyone can tell, but the type of bacteria that 'invades' or hitch-hikes into the mycelium and the only way to clean it is with antibiotic agar.
Let me illustrate what I'm talking about... Here it is a really good pic I took telling the difference. I took the nastiest jar for the photo. You can see some white 'branches' between the mycelium or even against the glass.  That's the bacteria I'm talking about and gotta pay close attention because it can go undercover, specially if you are managing lots of jars at the same time :rofl:. Sometimes it won't be as easy to spot, so you gotta keep close attention to it.
Hope it helps for the next runs!




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Edited by gabbk (09/03/20 06:46 AM)

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Re: Tomatose myc on agar, safe to transfer to grain? [Re: gabbk]
    #26915303 -

wow ok. So the second jar is bacterial... I couldnt have told myself.


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looking for a sclerotia producer print

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Re: Tomatose myc on agar, safe to transfer to grain? [Re: A.k.a]
    #26915359 -

A.k.a said:
Tomentose is fine, but I don’t like those plates. Something just looks wrong to me.

This is the most tomentose culture I’ve ever had.



Good to know; my melmac looks identical.

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Re: Tomatose myc on agar, safe to transfer to grain? [Re: otsilla]
    #26916923 -

Well, the myc hasn't grown through the agar yet, but I can see it growing between the sandwitch. Hope that one of them peeks through soon.

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Re: Tomatose myc on agar, safe to transfer to grain? [Re: otsilla]
    #26924289 -

So, yesterday it finally grew through, I transferred it immediately. Still appearing transparent at first, with some white stuff going on, as before, so I transferred  the leading part, scraped off a bit with inoculation loop so we'll see how it goes.

Here you can see the part where I scraped off.


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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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