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Can't clean up cultures on agar
    #26082778 -

I've been having a hard time getting a clean culture. These cultures were given to me and I used them for about 2 years without many problems, and I could always get clean organized flat rhizomorphic growth on my plates. It seems that there is a mold enmeshed into the cube mycelium that I can't transfer away from. I've tried several well timed transfers (about 10), hot pour, scraping little bits of the rhizo growth that's furthest from the center and appears to be clean, and none of it has led to a nice looking plate. I'm feeling a bit frustrated about this fuzzy white death on all my plates  :mad2:

My agar recipe is 7.5g LME, 10 g agar, and 500mL H20. These plates had a little less LME in them as I was trying to encourage fast rhizo growth. It didn't seem to help. I use a flow hood and hardly ever see contams outside of the weirdness infused into the culture on all my plates, and I hardly ever see a grain jar contam. Although I know often you don't see jars contam until its in a tub. All the fuzzy shit grew right up through the hot pour on every plate so it leads me to believe this is mold. I'd like to know what is this? Are there any other techniques for cleaning this up on agar? And if I clone a piece of the cap will contams like this often follow the myc all the way up into the mushrooms? I'm planning on trying to clone a stem and a cap soon anyway but any tips or feedback would be helpful.






This is how it began a few months ago. My plates went into the fridge looking clean and came out looking like this



I understand that cultures can go back and forth from rhizo to tomnetose based on nutrition and whatever but these are weird and most are disorganized and they all used to be rhizo on the same agar recipe I'm still using. Besides I keep hitting contams at the end of the 1st flush or some point in the 2nd flush using the same method that used to get me through 3 flushes no problem and often more.


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"When you pass through the waters, I will be with you; And through the rivers, they shall not overflow you. When you walk through the fire, you shall not be burned, Nor shall the flame scorch you."

Edited by Sherlock Shrooms (06/30/19 03:54 PM)

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Re: Can't clean up cultures on agar [Re: Sherlock Shrooms]
    #26082785 -

I've started new stuff from spores hopefully that goes well. My guess as to what caused this is that I was using an alcohol lamp in my flow hood and had a glass rectangular vase upstream from it to block the air, which was stupid. I wasn't getting the blade glowing orange, which was also stupid. I think that contams were blowing off the vase and onto my scalpel before I brought it into each dish. What I don't understand is why I have mold in my cultures and not bacteria, unless this is and I'm wrong about it. Because the scalpel was still hot even though it wasn't glowing and I imagine it would of scorched any molds but not been hot enough to kill bacteria. I don't know, I'm stumped


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"When you pass through the waters, I will be with you; And through the rivers, they shall not overflow you. When you walk through the fire, you shall not be burned, Nor shall the flame scorch you."

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Re: Can't clean up cultures on agar [Re: Sherlock Shrooms]
    #26082827 -

sometimes I get molds without bacteria when I make a mistake in sterile tek. just the way it goes i think.

I can't think of a method to get a clean piece that you didn't list. But I think I have an alternate idea. What if you tried to swab it? Swab across some cleanish myc hoping you pick up some bits of mycellium, then zig zag on a receiving plate like you would spores to try and separate things out a bit. Maybe, just maybe, you can get some clean bits on the receiving plate.


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Re: Can't clean up cultures on agar [Re: Sherlock Shrooms]
    #26082845 -

Starting new cultures from spores is probably your best option. If you've been continuing that culture for 2 years it could be senescing. That'll cause a loss of rhizomorphic growth and make them more vulnerable to contamination.


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Re: Can't clean up cultures on agar [Re: Kizzle]
    #26082849 -

Ya know, the more I looks at picture number 2 left culture, I can't see any immeshed molds or bacteria(maybe slight bacteria on the bright bit) Are you sure you're not just confusing Tomentose growth for mold. There's nothing wrong with Tomentose growth, they can produce every bit as well as Rhizo. Maybe you should throw a piece into a test grain jar and see what happens.

I also hear changing your nutrient can help reinvigorate an old culture.


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Re: Can't clean up cultures on agar [Re: LtLurker]
    #26082856 -

That's a pretty nifty idea. OP if you try this please let us know if it works. Assuming the mold is actually intermeshed and not just spores coming off that vase. I stopped messing with my alcohol lamp and started busting out the torch. Makes things a little easier/quicker especially with larger transfer sessions. Just remember to resanitize your hands after handling the torch. May be overkill but doesn't hurt just in case your hand accidentally crosses over the plate. And it keeps you occupied while the scalpel cools which helps it feel like an even quicker process. Hope you have success with the culture. Starting a fresh culture is a good idea though. Hell that's part of the fun :biggrin:


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Re: Can't clean up cultures on agar [Re: LtLurker]
    #26082859 -

Try a tiny tiny transfer from the best and cleanest area onto a new medium with peroxide.  H2O2 has worked miracles for me.  But if your culture is old then staring over from spores might be a good idea. But of course don't use peroxide with spores, only myc.


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Re: Can't clean up cultures on agar [Re: Sherlock Shrooms] * 1
    #26082870 -

You can use this system and It's just not for agar to LC. only.. You can use It for transferring from agar to agar.

I'm not saying It will help you and your situation but It's a good method to know.. I use the method at times..

The Biopsy Method: A different way of doing LC

https://www.shroomery.org/forums/showflat.php/Number/24740168/fpart/1/vc/1


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Re: Can't clean up cultures on agar [Re: LtLurker]
    #26082932 -

LtLurker said:
sometimes I get molds without bacteria when I make a mistake in sterile tek. just the way it goes i think.

I can't think of a method to get a clean piece that you didn't list. But I think I have an alternate idea. What if you tried to swab it? Swab across some cleanish myc hoping you pick up some bits of mycellium, then zig zag on a receiving plate like you would spores to try and separate things out a bit. Maybe, just maybe, you can get some clean bits on the receiving plate.


Good to know, and thanks for the tip. I'll try that. I'm not going to get my hopes up because whatever I'm tracking plate to plate is a fuzz that's on top of the myc and would likely come with the swab. Although if I just swab a little rhizo piece at the edge it might work. I'll give it a shot


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"When you pass through the waters, I will be with you; And through the rivers, they shall not overflow you. When you walk through the fire, you shall not be burned, Nor shall the flame scorch you."

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Re: Can't clean up cultures on agar [Re: Kizzle]
    #26082939 -

Kizzle said:
Starting new cultures from spores is probably your best option. If you've been continuing that culture for 2 years it could be senescing. That'll cause a loss of rhizomorphic growth and make them more vulnerable to contamination.


Thanks, that makes sense. So when they go senescent the culture itself will tend towards tomentose and sort of shoot out bits of rhizo growth like what I'm seeing on some of the plates? I should've started some new spores long ago but have been slacking. I think it's part of the teaching in what I'm experiencing


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"When you pass through the waters, I will be with you; And through the rivers, they shall not overflow you. When you walk through the fire, you shall not be burned, Nor shall the flame scorch you."

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Re: Can't clean up cultures on agar [Re: LtLurker]
    #26082945 -

LtLurker said:
Ya know, the more I looks at picture number 2 left culture, I can't see any immeshed molds or bacteria(maybe slight bacteria on the bright bit) Are you sure you're not just confusing Tomentose growth for mold. There's nothing wrong with Tomentose growth, they can produce every bit as well as Rhizo. Maybe you should throw a piece into a test grain jar and see what happens.

I also hear changing your nutrient can help reinvigorate an old culture.


I think you may be right about number 2 on the left. All I've been grabbing are little rhizo sections and I assumed a little rhizo tip I scraped off would create rhizo growth on the next plate. Maybe I did clean some of these up but mistook new clean growth for pure mold. I have some PDA maybe I'll make a batch and see what it looks like. I'll also drop some of the organized tomentose growths into grain jars and report back


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"When you pass through the waters, I will be with you; And through the rivers, they shall not overflow you. When you walk through the fire, you shall not be burned, Nor shall the flame scorch you."

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Re: Can't clean up cultures on agar [Re: Sherlock Shrooms]
    #26082976 -

Those cultures are odd for sure. When you spawn jars from them you still get fruits?

I've been stressing over meshed mold recently also. Got plates that look
beautiful and rhizo from the top, but if I look at them from the side there
are timy strands of myc standing up towards the lid. Happens in all my plates,
but my grow room is pretty warm also, so :shrug:


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Re: Can't clean up cultures on agar [Re: micelio]
    #26082979 -

KapnDank said:
That's a pretty nifty idea. OP if you try this please let us know if it works. Assuming the mold is actually intermeshed and not just spores coming off that vase. I stopped messing with my alcohol lamp and started busting out the torch. Makes things a little easier/quicker especially with larger transfer sessions. Just remember to resanitize your hands after handling the torch. May be overkill but doesn't hurt just in case your hand accidentally crosses over the plate. And it keeps you occupied while the scalpel cools which helps it feel like an even quicker process. Hope you have success with the culture. Starting a fresh culture is a good idea though. Hell that's part of the fun :biggrin:


I did start using a torch and you're right I have to sanitize my hands between each time using it but its not so bad. I have a jar of alcohol to dip my fingers into and rub them which makes it easy. You can see the torch suspended on the lower left hand side of my flow hood
Also don't mind me guys I just realized I can quote and reply to everyone in one text hehe
1WithU said:
Try a tiny tiny transfer from the best and cleanest area onto a new medium with peroxide.  H2O2 has worked miracles for me.  But if your culture is old then staring over from spores might be a good idea. But of course don't use peroxide with spores, only myc.


I didn't think h2o2 would survive the heat of pressure cooking?
micelio said:
You can use this system and It's just not for agar to LC. only.. You can use It for transferring from agar to agar.

I'm not saying It will help you and your situation but It's a good method to know.. I use the method at times..

The Biopsy Method: A different way of doing LC

https://www.shroomery.org/forums/showflat.php/Number/24740168/fpart/1/vc/1


Thanks I'll definitely check that link out when I get a chance.

Thank you everyone! I just realized that a few of these I did clean up but they reverted to total tomentose growth. Like a dumbass earlier today I threw a bunch of the plates that I felt I was hopelessly transferring into the trash :facepalm3: Many of them were still in sandwich bags and/or wrapped in cling wrap though :grin: I literally just went digging through the garbage in my garage to rescue a few haha. I'll wipe them down twice with alcohol before making any transfers  :rockon:


--------------------
"When you pass through the waters, I will be with you; And through the rivers, they shall not overflow you. When you walk through the fire, you shall not be burned, Nor shall the flame scorch you."

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Re: Can't clean up cultures on agar [Re: natedawgnow]
    #26082988 -

natedawgnow said:
Those cultures are odd for sure. When you spawn jars from them you still get fruits?

I've been stressing over meshed mold recently also. Got plates that look
beautiful and rhizo from the top, but if I look at them from the side there
are timy strands of myc standing up towards the lid. Happens in all my plates,
but my grow room is pretty warm also, so :shrug:


I do still get fruits. Lucky me! They don't make it nearly as many flushes as they used to though. Some turn green at the end of the first flush. I think Kizzle was right though that the ones that are a few years old may just be sensecent and succumbing to contamination sooner than they used to. That's sounds strange about the myc standing up toward the lid in all your plates. Has it turned to problems after spawning or have you not rolled with it and taken them that far since seeing that?


--------------------
"When you pass through the waters, I will be with you; And through the rivers, they shall not overflow you. When you walk through the fire, you shall not be burned, Nor shall the flame scorch you."

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Re: Can't clean up cultures on agar [Re: Sherlock Shrooms]
    #26082998 -

I am literally having the same exact problem. With the same white fuzz. I just created an account today because I became so frustrated. If you care to read it, here is my post addressing the same issue:

https://www.shroomery.org/forums/showflat.php/Number/26082594

I have also tried a hot pour that I didn't mention in my post AND I tried Josex' s biopsy method.

Fuq da fuzz man, it's ruining lives out here.

Mine can't be from senescence either because I keep going back to spores after 3-5 transfers and no progress.

I am new to agar so I was hoping it would turn out to be an odd nute issue or excess condensation.

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Re: Can't clean up cultures on agar [Re: MyNameIsntTrashcan]
    #26083073 -

Quote:
Fuq da fuzz man, it's ruining lives out here.


:hehehe: So true man. Sorry to hear about your troubles. I'll look at your thread and see if I have anything to offer, hopefully some folks can point you in the right direction.


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"When you pass through the waters, I will be with you; And through the rivers, they shall not overflow you. When you walk through the fire, you shall not be burned, Nor shall the flame scorch you."

Edited by Sherlock Shrooms (06/30/19 07:08 PM)

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Re: Can't clean up cultures on agar [Re: Sherlock Shrooms]
    #26083188 -

Thanks man. Atm I'm taking in information and advice like a black hole.

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Re: Can't clean up cultures on agar [Re: MyNameIsntTrashcan]
    #26083284 -

:takingnotes:


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"When you pass through the waters, I will be with you; And through the rivers, they shall not overflow you. When you walk through the fire, you shall not be burned, Nor shall the flame scorch you."

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Re: Can't clean up cultures on agar [Re: Sherlock Shrooms]
    #26084514 -

Just found this thread where Pasty chimes in and says it's definitely mold. In the OP CombackKid posts pictures that make this fuzzy growth easier to see. Maybe Sherlock can let us know if the fuzz on top of his myc shares the same visual characteristics as CombackKid's.

https://www.shroomery.org/forums/showflat.php/Number/23898121#23898121

Thank Todd some of y'all are decent with a camera.

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Re: Can't clean up cultures on agar [Re: MyNameIsntTrashcan]
    #26085977 -

It's hard to tell, the top pic in comebackkids post looks pretty wavy to be cube myc unless some strains do that, I've never seen it. A pic from the side may be more helpful.


This pic on the right is best obvious example I can offer, see the small sections that get a bit fuzzy. I'm pretty certain that is mold on the culture.


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"When you pass through the waters, I will be with you; And through the rivers, they shall not overflow you. When you walk through the fire, you shall not be burned, Nor shall the flame scorch you."

Edited by Sherlock Shrooms (07/02/19 11:02 AM)

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