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Invisibleb plus

Registered: 10/13/12
Posts: 928
Pan Cambo or Possible contamo
    #18178147 - 04/27/13 07:33 PM (11 years, 24 days ago)

Does this look like healthy, contam-free Pan cambo myc to you?



About to drop on wbs unless anybody has anything negative to say. This myc is fucking terrifying looking. Bottom left is the first plate I nocked with MS syringe I received in a trade. It was unused and from a reputable vendor, so I figured it was clean. Other two plates were nocked with transfers from bottom left plate. I've never seen mycelium like this before. Fast, cottony, cobweb-like growth. There is a strange yellow hue. IDK, scares the shit out of me. What do you guys think?


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:mushroom2:-Reishi Grow-:mushroom2:
:mushroom2:-Grey Oyster Grow-:mushroom2:

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Invisibleb plus

Registered: 10/13/12
Posts: 928
Re: Pan Cambo or Possible contamo [Re: b plus]
    #18178164 - 04/27/13 07:37 PM (11 years, 24 days ago)

:facepalm:

Can I please get this moved over to the contamination forum..


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:mushroom2:-Reishi Grow-:mushroom2:
:mushroom2:-Grey Oyster Grow-:mushroom2:

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OfflineTmethylM
Smear in the shale
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Registered: 07/16/12
Posts: 16,431
Loc: Florida
Last seen: 1 year, 7 months
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Re: Pan Cambo or Possible contamo [Re: b plus]
    #18178348 - 04/27/13 08:23 PM (11 years, 24 days ago)

Yeah thats pretty rough. Most of them don't really show a good growth pattern, and the mycelium is disorganized.
Was your agar media mixed properly? With the correct amount of ingredients?
Your mycelium in all of those doesn't look happy.

Despite the discoloration on the left plate, I would trust that one the most, it has very fluffy mycelium, and all Panaeolus species have a very fluffy mycelium.

I would transfer some wedges into a few WBS jars (enough for one tray or tub) and test it out before going bulk and inoculating a ton of jars.


Here is some Panaeolus mycelium



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¯\_(ツ)_/¯

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Invisibleb plus

Registered: 10/13/12
Posts: 928
Re: Pan Cambo or Possible contamo [Re: Tmethyl]
    #18178373 - 04/27/13 08:29 PM (11 years, 24 days ago)

Agar media:
600ml hpoo tea(boiled horse dung in pouches)
20grams agar powder
10grams LME

Filtered into glass pour bottle. PC'd for 90min at 15psi. Shook before I poured the plates to re-distribute any ingredients that may have begun to separate.

So I should avoid the plate on the right and the top? My biggest concern was the yellow/orange discoloration on the original ms plate. It seemed that the agar was almost "stained" a darker orange by the mycelium. When I transfered wedges the bottom of the wedges (uncolonized) was a dark orange color. I thought it may have been bacteria contamination, but it does not resemble anything I have ever seen before. I also noticed what you mentioned- the disorganized growth. ON the bottom right plate, mycelium from one segment of the dish grew up and over the barrier and over top of the other mycelium as a stratified layer. wtf?

If it is not Pan cambo mycelium on the bottom right and top plate than what is it?


--------------------
:mushroom2:-Reishi Grow-:mushroom2:
:mushroom2:-Grey Oyster Grow-:mushroom2:

Edited by b plus (04/27/13 08:34 PM)

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OfflineTmethylM
Smear in the shale
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Registered: 07/16/12
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Loc: Florida
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Re: Pan Cambo or Possible contamo [Re: b plus]
    #18178386 - 04/27/13 08:33 PM (11 years, 24 days ago)

Yeah just remove small wedges from the fluffy areas, the orange is likely not an issue though, it's partially digested agar and metabolites.
Doesn't look like bacteria.

Also, you do not need a huge wedge to colonize a jar, a wedge the size of a single rye grain is plenty IME.
Bigger wedge = more contams chance


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¯\_(ツ)_/¯

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Invisibleb plus

Registered: 10/13/12
Posts: 928
Re: Pan Cambo or Possible contamo [Re: Tmethyl]
    #18178423 - 04/27/13 08:40 PM (11 years, 24 days ago)

Thanks so much for the feedback:thumbup: You might have just saved my first pan grow. Digested agar/metabolite. That's pretty cool- defiantly better than what I was expecting to hear.

Why would there be contams present on the plates I made transfers onto but not the original MS plate? My sterile procedure was consistant. Plates only opened for split second to drop wedge. Multiple species worked on that day. ONly these two plates show this wispy type of growth. I also have 3 more pan cambo plates in the fridge that were nocked with the same ms plate. All of them have the same type(s) of growth. So if the bottom right and top plate pic in the picture is contamed- it's comming from the master ms plate? :shrug:

What agar recipe would you reccomend for future work with dung lovers? While were at it what general agar recipe would you recommend to use for most woodlovers?


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:mushroom2:-Reishi Grow-:mushroom2:
:mushroom2:-Grey Oyster Grow-:mushroom2:

Edited by b plus (04/27/13 08:59 PM)

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OfflineTmethylM
Smear in the shale
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Registered: 07/16/12
Posts: 16,431
Loc: Florida
Last seen: 1 year, 7 months
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Re: Pan Cambo or Possible contamo [Re: b plus]
    #18178860 - 04/27/13 10:25 PM (11 years, 24 days ago)

It's likely that none of them are contaminated, I see no mold or bacterial slimes anywhere.
I honestly think your agar mix was off somehow, the pan mycelium looks extremely weak in most areas.
Something is not right, could be moisture content too.

I'll let you in on my secret. I no longer use LME, ELME, or any type of agar really, I don't even use agar powder much anymore.

I grind up rye grains into a powder using a coffee grinder, mix with some pectin or agar powder(about 4 tblspn per qt).
Drop in some black dye, or whatever color I feel like, mix in some water very well, it should be runny, not thick or lumpy.
You'll have to experiment with it a little.
Now dump some into dishes, or jars, whatever I have handy, PC for 30mins @ 15psi.
Using this method I've germinated countless species of mushroom spores, wild collected and whatnot. I've cloned mushrooms, from grows and I've cloned mushrooms from grocery store, and I've done many strain isolations.

I've been doing this for a while now, there are some serious benefits here, which are overlooked by most:
Firstly, agar doesn't have to be clear, that is useless. You can spot contams on any color agar, even a solid color. I like to dye my rye agar black, because the mycelium, and any contams contrast extremely well, bacteria also contrasts well.

Secondly, it's cheap.. extremely cheap, and it's always on hand. If I contam a bunch, I really don't care. It's easy to clean up and pour some more.
I actually prefer 1/4pint or 1/2pint jars to work with the rye agar, because I keep breaking my glass petris.

Thirdly, you can store a bunch of premade rye agar in the fridge/freezer, you do not sterilize it like you would with LME agar.
YOu simply pour it into a plate/jar, then sterilize the plates/jars, this is best IME, sometimes the 'sterile' plates they sell aren't always sterile.
This method is thorough.

LASTYLY, and this is my favorite part! When you use a rye wedge, to inoculate a rye jar, there is no recovery/egocnition time, it grows right away and starts colonizing grains, because it already recognizes the material. Ever notice that dropping an LME wedge into a rye jar takes a few days for the mycelium to learn how to colonize the grains? It was already used to colonizng LME, and it can take a few days for it to recognize a new media.
Using rye wedges to inoculate rye jars saves a few days of colonization times, I am not exaggerating.

Also, rye agar works for woodlovers and all other saprotrophs.
I'll be writing this all up at some point, I'm keeping track of my projects right now, and will have a great visual guide soon.


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¯\_(ツ)_/¯

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Invisibleb plus

Registered: 10/13/12
Posts: 928
Re: Pan Cambo or Possible contamo [Re: Tmethyl]
    #18213026 - 05/04/13 12:31 PM (11 years, 17 days ago)



I will try this, and post results whenever you get around to making your thread. I've thought about doing the same thing with rye berries after seeing the success with BRF agar. Should have known someone already had it all figured out.


So one week after nocking the jar here it is:



What do you think?


--------------------
:mushroom2:-Reishi Grow-:mushroom2:
:mushroom2:-Grey Oyster Grow-:mushroom2:

Edited by b plus (05/04/13 12:36 PM)

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