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OfflineGloveboxFun
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Is this mold or oyster myc?
    #23773511 - 10/26/16 02:37 PM (7 years, 4 months ago)

Hi guys,
Long time listener, first time caller. I recently found a beautiful pleurotus cornucopiae specimen in the wild while hiking. I brought it home and transferred it to plates, but I'm not sure if I'm breeding mold or myc.

all of the plates have black spores in the sample, and two of the three have gotten a bacterial infection.
I'm not sure but one of the plates looks like it is sectoring, maybe its the bacteria. Do any of the below plates appear to have myc?

TLDR: do these plates have mold or oyster myc?





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Offlinekatbusa
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Re: Is this mold or oyster myc? [Re: GloveboxFun]
    #23773601 - 10/26/16 03:09 PM (7 years, 4 months ago)

Looks like a mixture of Oyster Myc, mold and bacteria. I would toss those. If those plates are glass PC them before you clean them.

Little Tip. Take a pic of each plate with light shining through it so we can see how the mycelium is segregating.

Actually hold off on tossing them just yet... Give us some info on what you did here.

What type of agar are you using?

How did you knock up the plate?

How long till you saw first growth?

How long did it take the plate to be covered in mycelium?

When did the watery globs show up?

Did you use a still air box or laminar flow hood?

The reason why I'm asking is because I think the first and second picture can be rescued if it is indeed oyster myc. Can you please give us a picture of the plate held up to a light source so we can see the mycelium. Like this:



Edited by katbusa (10/26/16 03:17 PM)

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OfflineGloveboxFun
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Re: Is this mold or oyster myc? [Re: katbusa]
    #23774909 - 10/26/16 09:51 PM (7 years, 4 months ago)

I used 2g agar and 5g malt powder (both from the international food store, scoured for malt extract but no luck) in 200ml water.
I tore the fruit down the middle and took a sample by scraping, and a sample was also taken from the gills. These were first transferred to mini mason jars containing agar. Most of the samples were contaminated, but I transferred what i thought was myc growth to the three plates. It took 2-3 days to start, and its been about 10 days so far. The watery globs appeared at 4 days and have gotten larger. This was done in a glove box which was sterilized with a 10% bleach solution.





I've had a bit of success with agar before but this has me completely confused.

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Offlinekatbusa
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Re: Is this mold or oyster myc? [Re: GloveboxFun]
    #23780706 - 10/28/16 05:15 PM (7 years, 4 months ago)

So normally you would transfer good clean mycelium to another agar plate pretty soon after the good and bad mycelium start to expand.

I'm pretty sure the more fuzzy mycelium is the oyster myc. Take a small sample from that and watch it expand again. Take samples from the leading edge of the good oyster mycelium. Keep replating till the culture is clean. You do this for both mold and bacteria.

The only way to control mold is to give the oyster mycelium a low nutrient environment. This will cause the mycelium to run faster and hopefully out run the mold so you can get a clean sample from the leading edge. A good recipe is:

10 grams agar
500ml water
3 to 4 grams of malt extract

If you are having a hard time finding malt extract take a look at some of the Agar recipes here. You should be able to find one that will work for what you have available. Just cut the required nutrients in half or a little below half.

https://www.shroomery.org/5243/What-are-some-good-agar-media-recipesust cut the required nutrients in half or a little below half.


Now to clean the bacteria. Bacteria rods can be present on growing mycelium. Thankfully bacteria is easier to isolate from. You want to use an antibacterial agar. The antibacterial agar can be created two ways. The first way is by using gentramicin @ 100mg per liter of agar or try tea. I personally suggest using tea because it's organic and cleaner. Antibacteria Tea Agar is still in the research stage but it does work. I have used it to clean a pretty dirty culture. Here is a thread about Antibacterial Tea agar.

https://www.shroomery.org/forums/showflat.php/Number/23771470

Below is a recipe you can use:

3 non-favored Green Tea bags
1 Liter of water
10 grams of agar
2 grams of malt extract

Add the tea bags to the water before you add any thing else. Let the tea bags steep in the hot water for 5min. Remeasure the water and add more water to bring it back up to 1 Liter. Then add the other ingredients and PC as you normally would. 

Please come back to this thread and update us on how it worked for you!!

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OfflineGr0wer
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Re: Is this mold or oyster myc? [Re: katbusa]
    #23781718 - 10/29/16 01:53 AM (7 years, 4 months ago)

Did you backlight those dishes? The light makes it hard to see anything. Unless you have crystal clear agar we want the lighting from the front.

Edited by Gr0wer (10/29/16 01:54 AM)

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Offlinekatbusa
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Re: Is this mold or oyster myc? [Re: Gr0wer]
    #23781945 - 10/29/16 07:16 AM (7 years, 4 months ago)

He has both from the front and back lighting. I asked for back lighting because I wanted to see if there was any major density extremes in the mycelium incase he was going to ask where to take a sample. Plus sometimes you can see bacteria that has been covered up by the myc.

Edited by katbusa (10/29/16 07:23 AM)

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OfflineGloveboxFun
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Re: Is this mold or oyster myc? [Re: katbusa]
    #23813615 - 11/08/16 06:17 PM (7 years, 3 months ago)

Thank you katbusa for the help!

Update;
I took slivers of several of the fuzzier looking parts and transferred them onto clean plates.



Four got the same black shiny mold that was present in the earlier dishes, but one escaped clean. I also noticed that the outermost growth on all the plates was very thin, very unlike cubensis growth.



My clean culture (sorry about the condensation, poured too hot)



I then took this clean culture and used the sandwich method and some antibiotic black tea agar to clean it of bacteria (this recipe) To my amazement, two days after the transfer I noticed mycelium poking through the top, this is the result today:





I also did a few straight transfers to compare the growth of the sandwich transfers.





Thanks again for all the great input, I'm still new to this hobby but I am very happy that there is a place where I can learn more.

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OfflineFerather
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Re: Is this mold or oyster myc? [Re: GloveboxFun]
    #23815440 - 11/09/16 07:55 AM (7 years, 3 months ago)

Well done with the tea agar clean. Good job, it is very easy to do.
I am using T-Gel, which is my 100% starch free version.

Nothing to pressure cook, even easier to do.


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

Growing mushrooms, general guide and information (Ferather's Journal), https://ibb.co/rG3rML2

https://www.shroomery.org/forums/showflat.php/Number/27857366#27857366

DTS DCH Driver for Realtek [DTS:X] - Unlocked and reprogrammed.

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OfflineGloveboxFun
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Re: Is this mold or oyster myc? [Re: Ferather]
    #23835699 - 11/15/16 02:43 PM (7 years, 3 months ago)

Thanks Ferather, I appreciate the good vibes. Ill have to try that tek out next time i clone.

UPDATE

Here is the current state of the mycelium.


The triangle wedge is in the top right in the above image.


The square wedge is in the top left in the above.
Both of the wedge transfers have slight yellow discoloration in a few spots, not sure what to make of it, the other plates are supa-dupa-clean.


Fuzzier than drone metal bass.
The wedge transfers are more circular in appearance, but the sandwiched myc is definitely more fluffy and has more variation in appearance.

Would it be ok to transfer these to grain now or should i do more transfers to get a really isolated strain? If so, how do i know which section to pick?

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OfflineFerather
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Re: Is this mold or oyster myc? [Re: GloveboxFun]
    #23835816 - 11/15/16 03:22 PM (7 years, 3 months ago)

Awesome job, I'm jealous you have Pleurotus Cornucopiae.

Well done, nice clean clone.  :thumbup:


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

Growing mushrooms, general guide and information (Ferather's Journal), https://ibb.co/rG3rML2

https://www.shroomery.org/forums/showflat.php/Number/27857366#27857366

DTS DCH Driver for Realtek [DTS:X] - Unlocked and reprogrammed.

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Invisibleflyontoast
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Re: Is this mold or oyster myc? [Re: katbusa]
    #23838726 - 11/16/16 12:43 PM (7 years, 3 months ago)

Quote:

katbusa said:
He has both from the front and back lighting. I asked for back lighting because I wanted to see if there was any major density extremes in the mycelium incase he was going to ask where to take a sample. Plus sometimes you can see bacteria that has been covered up by the myc.



What does it mean if there are "density extremes"?


--------------------
:mushroom: My trade list :mushroom:
Looking for strong terrestrial fruiters for an outdoor beds experiment:
Agaricus Bitorquis, Agaricus Augustus, Agaricus blazei/subrufescens, Stropharia Rugoso-annulata, Clitocybe Nuda (blewits), and any species or other genus that you think work outdoors.
Also, any commercially viable Pleurotus, cold or hot strains.
Thanks for the Q&A, trades, and all the posters & teachers that have come before us  :skol:

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OfflineGloveboxFun
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Re: Is this mold or oyster myc? [Re: flyontoast]
    #23872337 - 11/27/16 02:56 PM (7 years, 3 months ago)

Ferather,
Im fairly certain its p. cornucopiae, it was a single white trumpet shaped fruit growing from the side of a log, and the guidebook i have suggested thats what it might be. It might also end up being a different variety, only fruits will tell

fly,
I think the density of the growth can determine if it is mycelium or mold, especially cobweb because of how wispy it can get.

UPDATE
Growth has gotten pretty intense, the whole plate is overgrown




I took a few wedges from a few of the good looking plates and moved them to grain





Within a week they fluffed up pretty nice, looked very thin at first and then began looking more like mycelium, shook them when they were about a third of the way colonized to spread out the colonization points.
Im excited to spawn to straw and fruit these!

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OfflineFerather
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Re: Is this mold or oyster myc? [Re: GloveboxFun]
    #23872394 - 11/27/16 03:14 PM (7 years, 3 months ago)

Nice, when you get to fruiting, it will make a tuber, and then caps. Looks like a knobbly truffle.
When the fruit body's fully develop properly they will branch from one single base.

Here is a weblink for: Pleurotus cornucopiae (Branched oyster).


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

Growing mushrooms, general guide and information (Ferather's Journal), https://ibb.co/rG3rML2

https://www.shroomery.org/forums/showflat.php/Number/27857366#27857366

DTS DCH Driver for Realtek [DTS:X] - Unlocked and reprogrammed.

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