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Invisiblercraft
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Registered: 02/06/22
Posts: 37
Sanity Check - Agar Health
    #27785850 - 05/20/22 02:29 PM (2 years, 7 months ago)

Background:

Agar recipe is 10g LME 10g agar 500ml water. PC at 15 PSI for 15 minutes.


These plates are my second transfer. The original plate was started with a drop of MSS, and it grew from the center of the plate and looked very much like these.




I am not super concerned about getting strong rhizo growth from these today. I have a couple of clone plates going that have more rhizo growth, but they definitely need another transfer because there are pockets of bacteria.

But to some questions:

1. I am fairly sure this is just tomentose mycelium, but is it possible that I'm delusional and just growing mold? I have multiple plates that look like this (I took 4 transfers, all of them look like this), would opening one to give it a sniff rule out bacteria? My last grain jars were lost to some bacillus (strong sour smell, like rotting apples).

2. These plates are growing with good speed, they were transferred 5 days ago. Seems like a reasonable rate. I think that once the plate is fully occupied, I could take some slices to grain, and some slices to another transfer. Should I do more transfers before it makes sense to create spawn, though?

Thanks in advance!

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Invisiblercraft
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Registered: 02/06/22
Posts: 37
Re: Sanity Check - Agar Health [Re: rcraft]
    #27790999 - 05/24/22 06:49 PM (2 years, 7 months ago)

Update

I am now fairly sure that I have a bad case of Bacillus :smile: I guess it could be that there is mycelium there, but it's terribly contaminated with substantial bacillus based on the smell and the outcome of the last jars I had to toss.

Today I got to doing some transfers, and took the opportunity to smell test one of the plates since I had so many that looked the same. It smelled bad - a bit sour, like maybe fermenting apples.


---

So, I used my remaining 6 agar plates to take two transfers from a healthy piece of clone that had been attacked by bacteria on 3 sides, two transfers from my Bacillus plates (in case there is mycelium, I plan to do a fast transfer before it has the chance to sporulate and ruin it all), one with King Oyster spores, and one with some old B+ spores that I'm praying germinate because I'm out of tubs and almost out of spores :P

Fingers crossed that I'm on the path towards some healthy agar plates!

EDIT: Here is what the smelly plate looked like today.

I took transfers from two different places, probably should have stayed away from the fluffier parts but I am still learning.

Edited by rcraft (05/24/22 06:56 PM)

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InvisibleSlugWorthx1S
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Re: Sanity Check - Agar Health [Re: rcraft]
    #27791050 - 05/24/22 07:18 PM (2 years, 7 months ago)

Generally, bacterial plates would look wet, mold will produce fruiting bodies fairly quickly. Looks good from what i can see in first few images

Edit: if your plates omits any odor besides mycelium, id toss it.

Edited by SlugWorthx1 (05/24/22 07:28 PM)

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Invisiblercraft
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Registered: 02/06/22
Posts: 37
Re: Sanity Check - Agar Health [Re: SlugWorthx1]
    #27791092 - 05/24/22 07:45 PM (2 years, 7 months ago)

Quote:

SlugWorthx1 said:
Generally, bacterial plates would look wet, mold will produce fruiting bodies fairly quickly. Looks good from what i can see in first few images

Edit: if your plates omits any odor besides mycelium, id toss it.





Hm, interesting. That makes sense - I guess it's likely that there's bacteria lurking *under* that mycelium!

I think that I need to do faster transfers off clean edges more, before the bacteria can take control. Gotta get my next agar batch less wet, for sure - I struggle with condensation :smile: .

Thanks!

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InvisibleMTZ
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Re: Sanity Check - Agar Health [Re: rcraft]
    #27791110 - 05/24/22 08:06 PM (2 years, 7 months ago)

There's many molds that aren't wet. They'll grow white and fluffy looking. Just like mycelium to a point. That's what it looks to me. Maybe do some transfers like you should anyway and see if it cleans up. If not? It's no biggie. Just some time anyway. When I'm starting a new culture, on the LOW side I start no less than 5 plates. 10+ is the usual. That way I've got plenty to choose from and losing a plate isn't a big deal.


--------------------
Rehears your death every morning and night. Only when you constantly live as though already a corpse will you find freedom in the martial way, and fulfill your duties without fault throughout your life. ~ The Book Of Samurai
........
I live in the void. I can go to the in between anytime I desire. I visit the spirt world.

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Invisiblercraft
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Registered: 02/06/22
Posts: 37
Re: Sanity Check - Agar Health [Re: MTZ]
    #27805628 - 06/04/22 11:52 AM (2 years, 7 months ago)

Alright, I've been doing some ✨transfers✨! I also made a new set of agar plates very carefully and successfully ended up with 16 out of 20 not having any condensation. A combination of lower temp pour + putting a much thinner layer on each plate worked well for that (the last 4 are almost condensation free, too).

These are the 3rd transfer plates I sampled today (these had been growing for ~5 days):


- Really interesting "river" looking pattern, probably bacteria. The edges of the plates sometimes had a touch of visible water, probably a vector for that growth. I also wasn't super careful on the last transfer - the last plate was quite dirty.


- This one is pretty clean looking besides the one yellow bacterial colony. Except:

Quote:


There's many molds that aren't wet. They'll grow white and fluffy looking. Just like mycelium to a point. That's what it looks to me. Maybe do some transfers like you should anyway and see if it cleans up. If not?




It still kind of *smelled* a bit, faint - I couldn't tell if it was the agar fumes, or what - but it still makes me suspicious that I'm not actually transferring myc here. Hoping that in a plate or two it'll start showing more obvious rhizo growth.



And thankfully, I managed to rescue one of my clones! This plate is looking pretty exciting right now - growth rate is slower than the MSS ones that smell above, but it looks better:

- For 5 days of growth it seems a bit slow but not abnormal.

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InvisibleLadysKnight
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Re: Sanity Check - Agar Health [Re: rcraft]
    #27805861 - 06/04/22 03:33 PM (2 years, 7 months ago)

All are mold


--------------------
Don't follow leaders

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OfflinePBJ710
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Re: Sanity Check - Agar Health [Re: LadysKnight]
    #27805915 - 06/04/22 04:25 PM (2 years, 7 months ago)

:whathesaid:

Toss them and start again from spores.  Use a loop to streak the MS instead of just putting a drop on the plate.  This will spread the cultures out so you can easily choose healthy ones to transfer from.  When you take transfers, use tiny transfers so you'll be less likely to bring hidden contams over on it.

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OfflineSockadin
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Re: Sanity Check - Agar Health [Re: PBJ710]
    #27805937 - 06/04/22 04:52 PM (2 years, 7 months ago)

Yeah man they's all mold.

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Invisiblercraft
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Registered: 02/06/22
Posts: 37
Re: Sanity Check - Agar Health [Re: Sockadin]
    #27805947 - 06/04/22 04:59 PM (2 years, 7 months ago)

Ok, thanks guys. Are you also saying the clone in the last pic is just succumbed to mold? It seems a bit too thick and white to be something other than myc, but I am pretty new.

In any case, I took the last few drops of my syringe and looped it to 5 more plates. Hopefully something comes out of those!

Edited by rcraft (06/04/22 05:00 PM)

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OfflinePBJ710
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Re: Sanity Check - Agar Health [Re: rcraft]
    #27805994 - 06/04/22 05:54 PM (2 years, 7 months ago)

At best your initial transfers were meshed with the mold.  I would have tossed that T1 plate when I saw it grow out like that.  Unless there is something really special about a culture, it's normally not worth trying to seperate the two as you can just make new plates in much less time.

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Invisiblercraft
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Registered: 02/06/22
Posts: 37
Re: Sanity Check - Agar Health [Re: PBJ710]
    #27806163 - 06/04/22 08:34 PM (2 years, 7 months ago)

Quote:

PBJ710 said:
At best your initial transfers were meshed with the mold.  I would have tossed that T1 plate when I saw it grow out like that.  Unless there is something really special about a culture, it's normally not worth trying to seperate the two as you can just make new plates in much less time.




Thank you PBJ, I appreciate your patience and insights :smile:

Nothing special about this plate really - I'm mostly fascinated with figuring out what I am looking at! So like - yea, mold - I'm keeping them around to see how it plays out and get a better idea of what I'm seeing. At the same time, I have no shortage of ingredients, time, and stubbornness to "try again" - so here I am doing it all in parallel.

--

I am just getting past the "shit, I'm gonna run out before my next grow finishes" anxiety. It is now inevitable. With that, I can focus on improving my process and getting some security in my supply.

All of the assistance is greatly appreciated.

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Invisiblercraft
Stranger
Registered: 02/06/22
Posts: 37
Re: Sanity Check - Agar Health [Re: rcraft]
    #27810339 - 06/07/22 11:45 PM (2 years, 7 months ago)

Alright, so tonight I had some fun and tried out my new microscope :laugh:

I looked at a few plates, getting practice. I had one plate that had an old piece of mushroom on it from a botched clone attempt. So I'm pretty sure this is the mycelium I expect to see:



It was a bit clearer in the eypieces, but it looked pretty close to the first pic in this post https://www.shroomery.org/forums/showflat.php/Number/25975793#25975793

(ignore all of the dots, that was likely me being lazy and not using distilled water or doing really anything to keep it clean, whoops).

Then, I tried to figure out wtf was going on with the mold plates from earlier in the thread. it's definitely not mushroom myc - brushing it with a piece of tape lifted it ALL off!



Overall, it kind of looked more like... fiberglass than anything else. Couldn't even really make out a network - almost like it shattered when I picked it up. Or, I am just bad at doing microscope work. I have like 6 more mold plates to try out and see if I can't get a better picture.

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