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OfflineL1740
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Contam or bruising?
    #26591211 - 04/10/20 02:23 PM (3 years, 10 months ago)

Hi.

I am pretty new to mushroom cultivation and eager to learn. :smile:

I made several agar plates with spores. One of my plates started getting teal in the center of the mycelium. After some time it also developed several more rings, concentric to the center. I thought this was bruising and did not really mind it.

But my transfers are turning teal. I am wondering if this might be contam, if I am doing anything wrong or if it is just how this variety of mushroom is.

This is my first agar plate just before I started doing transfers from it. Teal center and rings:




These transfers have been done 1 day ago. They are looking promising, and I have been even more strict on the hygiene here:








These are some transfers that are around 2 days old. They are starting to show some teal color other places than center:






This sample is 4 days old and is turning more green. All my samples started out looking like the fresh ones above, with teal colors in some places, and then turned out like this. I am guessing this is contam:






A side note: I alco sterilize my SAB and then wash it with soap and leave the surfaces wet with soapy water. I sterilize all my jars with alcohol and flame sterilize my scalpel between each transfer. The latest two transfer (yesterday and the day before) I also took a thorough shower and washed the whole bathroom, which is where I do my transfers. I have done transfers of other varieties as well (Shiitake and Oyster) and they turn out clean, without any contam at all. Done in the same SAB during the same session. But my magical ones turn out teal, and eventually green. I think it is strange that all the transfers from my first jar (which I think looks ok) are turning all green like the last image after a couple of days.

As I said, I am new to this hobby, but I am enjoying it so far! Thanks for all the great information shared here so far :laugh:


Edited by L1740 (04/18/20 05:43 PM)


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Offlinerumfor69
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Re: Contam or bruising? [Re: L1740] * 1
    #26591330 - 04/10/20 03:25 PM (3 years, 10 months ago)

Definitely is a contamination. But the goal of agar is to cut the very
edge of outstretched mycelium to hopefully "clean up" the culture.
My suggestion would be to go back to the original dish and try again.
Only cut a very small piece about the size of a grain of rice from
the outer edge.


--------------------

Links For Beginners And Beyond:mspoil:


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OfflineL1740
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Re: Contam or bruising? [Re: rumfor69]
    #26591363 - 04/10/20 03:38 PM (3 years, 10 months ago)

Thanks for the answer rumfor69! Much appreciated. :smile:

I have tried to only transfer a small piece of the edge of the mycelium.

But the transfers I have that are just 1 day old. Should they show signs of contamination this early, or will it come later? They look very white and clean now.

And another question:
The two days old transfer. are the white edges mycelium, or just contam in an early stage?


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Offlinerumfor69
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Re: Contam or bruising? [Re: L1740]
    #26591425 - 04/10/20 04:03 PM (3 years, 10 months ago)

Once the contamination has germinated it can grow quickly, faster than
the mycelium even. And once its sporelating like this it might be
impossible to clean up.

The white edge in day 2 like you circled might be ok to try.

All this mycelium looks a bit off though and might not be able
to be cleaned up. I'm hoping some other eyes chime in here too.


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OfflineL1740
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Re: Contam or bruising? [Re: rumfor69]
    #26591469 - 04/10/20 04:19 PM (3 years, 10 months ago)

Ok. Bummer.

I will not give up though, will let it grow and observe, and try to do some small transfers from the cleanest looking whites. :smile:

When you say it looks off, what exactly do you mean by that? It is kind of cottony, and not rizomorph. I use a mix of 10g agar, 10g LME and 5dl water.


Edited by L1740 (04/18/20 05:44 PM)


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Offlinerumfor69
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Re: Contam or bruising? [Re: L1740]
    #26591622 - 04/10/20 05:28 PM (3 years, 10 months ago)

Yeah little fuzzy/cottony which might just mean it's tomentose mycelium
instead of rhizo. I've had lots of good luck with fuzzy mycelium. Don't
let any of them sit for very long they'll just sporelate if they're contaminated
and it will make it harder.


--------------------

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OfflineL1740
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Re: Contam or bruising? [Re: rumfor69]
    #26591628 - 04/10/20 05:30 PM (3 years, 10 months ago)

Noted :smile: Will prepare some new agar dishes for tomorrow.

Thanks man, much appreciated :smile:


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InvisibleRoger Clemency
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Re: Contam or bruising? [Re: L1740]
    #26591662 - 04/10/20 05:44 PM (3 years, 10 months ago)

I don’t know if you have any cube mycelium there at all. The little piece on the right in the 3rd pic looks like it could be but is entangled with trich or something like it.

That ringed first pic looks just like a plate of trich I was just checking out, though the trich was more green in the rings.

Are you out of cube spores?


--------------------
Sour grapes, sweet revenge
Heaven starts right where hell ends


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OfflineL1740
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Re: Contam or bruising? [Re: Roger Clemency]
    #26591762 - 04/10/20 06:23 PM (3 years, 10 months ago)

Hm. No mycelium at all? I'll do some more testing and waiting to find out.

Thanks for your view on this as well!


Edited by L1740 (04/18/20 05:46 PM)


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InvisibleRoger Clemency
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Re: Contam or bruising? [Re: L1740]
    #26591788 - 04/10/20 06:34 PM (3 years, 10 months ago)

I’m pulling for you man :mushroom2:

What country do you live in? There are some vendors of the website that are still shipping stuff


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Sour grapes, sweet revenge
Heaven starts right where hell ends


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OfflineL1740
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Re: Contam or bruising? [Re: Roger Clemency]
    #26591797 - 04/10/20 06:38 PM (3 years, 10 months ago)

Thanks.

I’ll post some updates when my current dishes develop in any direction and if I find new spores :smile:


Edited by L1740 (04/18/20 05:47 PM)


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InvisibleRoger Clemency
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Re: Contam or bruising? [Re: L1740]
    #26591821 - 04/10/20 06:46 PM (3 years, 10 months ago)

Roger that. I wish the world in general would realize how amazing these things are, it’s a fkin crime that someone can get kidnapped and held in a cell over some magical food that grows naturally. What have we done to ourselves...these  little guys in particular are probably what made humans into the self aware creatures we are today.


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Sour grapes, sweet revenge
Heaven starts right where hell ends


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OfflineL1740
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Re: Contam or bruising? [Re: Roger Clemency]
    #26591837 - 04/10/20 06:52 PM (3 years, 10 months ago)

Exactly, a shame! One day... :smile:


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OfflineL1740
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Re: Contam or bruising? [Re: L1740]
    #26593094 - 04/11/20 09:45 AM (3 years, 10 months ago)

I got a hold of some spore prints today. Will do first agar with new spores this evening. I'll do my best to keep thing sterile!


Edited by L1740 (04/18/20 05:48 PM)


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OfflineHamHead
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Re: Contam or bruising? [Re: L1740]
    #26593314 - 04/11/20 11:35 AM (3 years, 10 months ago)

Yeah, no.

Start over with spores to agar.

That first plate is completely fucked.



You want to see something similar to that.


--------------------
The Italian researchers’ findings, published by the INT’s scientific magazine Tumori Journal, show 11.6% of 959 healthy volunteers enrolled in a lung cancer screening trial between September 2019 and March 2020 had developed coronavirus antibodies well before February.

https://www.reuters.com/article/us-health-coronavirus-italy-timing-idUSKBN27V0KF

This online first version has been peer-reviewed, accepted and edited,  but not formatted and finalized with corrections from authors and proofreaders

https://www.icandecide.org/


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OfflineL1740
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Re: Contam or bruising? [Re: HamHead]
    #26600311 - 04/14/20 12:10 PM (3 years, 9 months ago)

Thanks for the feedback Hamhead. All old agar dishes are tossed away, and I have made 6 new agar plates with spores from 3 different prints. I also inoculated 4 jars of BRF, with spores from two of those same prints.

It has been sitting for 3 days now, and I am starting to see some tiny white stuff around my spores. I hope this is myc. I'll post updates as this grows. I intend to start isolating when I am getting some solid plates.


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OfflineL1740
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Re: Contam or bruising? [Re: L1740]
    #26607582 - 04/17/20 09:12 AM (3 years, 9 months ago)

Just a quick update. I made seven (not six) new agar jars, and so far all 7 show signs of growth. Two of them were made some days later, and they are just starting to develop as well. I am waiting for 40 petri dishes to arrive shortly, and I’m looking forward to moving over to a more compact and handy format! Also hope its easier to observe and take pictures of the mycelium than through the bumpy walls of my jars.

All 4 jars of brf are starting to show growth as well. So happy I just started over again. Thanks for the help so far ☺️




Edited by L1740 (04/17/20 09:18 AM)


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InvisibleRoger Clemency
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Re: Contam or bruising? [Re: L1740]
    #26608040 - 04/17/20 12:13 PM (3 years, 9 months ago)

Niiice! I’m glad you got some stuff going on man. You can transfer a little piece from any of those original spore plates now any time, just to make sure it’s clean. It’s kind of hard to tell what’s going on at first with spores because everything is so fluffy and kinda chaotic.

Get some nice growth, I’d make another transfer but also put a few wedges in grain jars. Then let a plate or two colonize all the way and see if they fruit, if they do grab a pin and transfer it. That will likely be a good performing clone.


--------------------
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Heaven starts right where hell ends


Edited by Roger Clemency (04/17/20 12:20 PM)


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OfflineL1740
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Re: Contam or bruising? [Re: Roger Clemency]
    #26608060 - 04/17/20 12:22 PM (3 years, 9 months ago)

Thanks man.

Ok, I was not aware og letting it pin in the dish and do a clone from here would result in a good performer, but I will definately try it out!! 👏🏼😁


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Offlinerumfor69
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Re: Contam or bruising? [Re: L1740]
    #26608485 - 04/17/20 04:05 PM (3 years, 9 months ago)

Nice! I'm glad you got some stuff going on. And starting over was
probably the best choice considering how things looked.


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