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Offlineesotericbound
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Panaeolus Cyanescens multispore growth on agar appearance?
    #28169274 - 02/02/23 02:45 PM (1 year, 1 month ago)

I'm hoping people experienced with pan cyans can tell me what I may be dealing with on these pan cyan agar plates.  My experience has been with cubes so far, and I generally have high success/low contam rate with my setup and methods, but these agar plates don't look right to me, especially the ones with more brown areas and droplets on the surface.  These are from Panaeolus Cyanescens Jalisco Mexico spore prints.







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"...If you have any kind of intellectual filter at all, you quickly can satisfy yourself that our best efforts are nothing more than half completed stories told around the campfire; we don't actually know what our predicament is."  -Terence McKenna

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OfflineAspectOfTheCreator
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: esotericbound]
    #28169413 - 02/02/23 04:11 PM (1 year, 1 month ago)

I'm somewhat new to agar so take what I say with a grain of salt. I got a good number of Pan Cambo plates going. You're correct in that those plates definitely don't look right. My pan cambo growth has all been thick, white, fluffy growth. Very cloud-like



On BRF agar







blue is potato flake/corn syrup agar, the clear agar is LMA. It's grown more or less the same on each.

Are those from transfers? MSS? Streaks?


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Edited by AspectOfTheCreator (02/02/23 04:12 PM)

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Offlineesotericbound
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: AspectOfTheCreator]
    #28169457 - 02/02/23 04:37 PM (1 year, 1 month ago)

These are from recently acquired spore prints, no streaks, literally scraped off the foil (seems to work as good/better than streaks ime).  The same time I did these, I also did some from pan cyan texas print, and those I think are fine (just white fluffy), which leads me to think there was contam on the jalisco print.  It's all done with big flow hood and torched scalpel, 70%IPA, etc.  I guess I'll try again from print, and also transfer from best looking of these plates.


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"...If you have any kind of intellectual filter at all, you quickly can satisfy yourself that our best efforts are nothing more than half completed stories told around the campfire; we don't actually know what our predicament is."  -Terence McKenna

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Invisibleruawakeyet
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: esotericbound]
    #28169477 - 02/02/23 04:47 PM (1 year, 1 month ago)

Where does the 70% IPA come in to play when using a FH and flame sterilization?

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Offlineesotericbound
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: ruawakeyet] * 1
    #28169484 - 02/02/23 04:52 PM (1 year, 1 month ago)

IPA just for sanitizing myself, work area, bags of the petri dishes before opening etc....


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"...If you have any kind of intellectual filter at all, you quickly can satisfy yourself that our best efforts are nothing more than half completed stories told around the campfire; we don't actually know what our predicament is."  -Terence McKenna

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Invisibleruawakeyet
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: esotericbound]
    #28169488 - 02/02/23 04:54 PM (1 year, 1 month ago)

Ok, just checking.

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OfflineAspectOfTheCreator
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: esotericbound]
    #28169538 - 02/02/23 05:32 PM (1 year, 1 month ago)

Try to reinoculate another dish and this time take a transfer ASAP rather than letting it grow out.


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Offlineesotericbound
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: AspectOfTheCreator]
    #28169576 - 02/02/23 05:56 PM (1 year, 1 month ago)

I think I understand the objective, and would work if mycelium was there first, but the contam appeared almost before the mycelium on these dishes.  I'll try though...


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"...If you have any kind of intellectual filter at all, you quickly can satisfy yourself that our best efforts are nothing more than half completed stories told around the campfire; we don't actually know what our predicament is."  -Terence McKenna

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InvisibleBaba Yaga
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: esotericbound] * 1
    #28169638 - 02/02/23 06:45 PM (1 year, 1 month ago)

Are you incubating these plates or keep at room temp?

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Offlineesotericbound
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: Baba Yaga]
    #28169658 - 02/02/23 06:59 PM (1 year, 1 month ago)

maintaining temp @79-80°F


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"...If you have any kind of intellectual filter at all, you quickly can satisfy yourself that our best efforts are nothing more than half completed stories told around the campfire; we don't actually know what our predicament is."  -Terence McKenna

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InvisibleBaba Yaga
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: esotericbound] * 1
    #28170233 - 02/03/23 03:15 AM (1 year, 1 month ago)

How are you maintaining the temperature? Are you heating the room or are you using a heat mat or similar?
I'm asking cause the first thing that popped into my mind when seeing your plates was that they are kept too warm.

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Offlineesotericbound
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: Baba Yaga]
    #28170453 - 02/03/23 08:14 AM (1 year, 1 month ago)

Interesting.  This is in a well insulated 8'x8' room with 600w heater at opposite side of room, regulated with separate temp controlled relay.  3 temp/humidity sensors in room recorded 78.3/81.5 min/max.  Does it look like a bacterial thing or just stressed out mycelium?  I'm wondering if there was something about the agar it didn't like.

I did some transfers from best looking sectors yesterday to agar with 1000ml water/8g MEA/4g potato starch/.4g peptone/ 18g agar, will see how they take


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"...If you have any kind of intellectual filter at all, you quickly can satisfy yourself that our best efforts are nothing more than half completed stories told around the campfire; we don't actually know what our predicament is."  -Terence McKenna

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Invisibleruawakeyet
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: esotericbound]
    #28170463 - 02/03/23 08:21 AM (1 year, 1 month ago)

I would simplify that agar recipe. Is that "8g MEA" premixed ME and A-A, or just ME? If it's premixed, plus the 18g agar agar powder, that is likely too hard.

For pans, (and practically everything else), plain ol' MEA works great. ME should be at 1% max, A-A should be as low as possible and still firm enough to take transfers.

These fancy recipes tend to make thicker growth that is more difficult to read on agar.

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Offlineesotericbound
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: ruawakeyet] * 1
    #28170474 - 02/03/23 08:30 AM (1 year, 1 month ago)

Gotcha, it's essentially the recipe jakeoncid lists sans yeast.  I meant 8g ME (dried malt extract light).  Will try simple 1% ME next round.


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"...If you have any kind of intellectual filter at all, you quickly can satisfy yourself that our best efforts are nothing more than half completed stories told around the campfire; we don't actually know what our predicament is."  -Terence McKenna

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Invisibleruawakeyet
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: esotericbound]
    #28170480 - 02/03/23 08:36 AM (1 year, 1 month ago)

That might be the only thing I disagree with Jake about. I tried a similar fancy recipe, and it was a nightmare for me. I've omitted all other ingredients and haven't looked back. The fanciest I get now is using different nutrient levels for different genuses/species.

Pans get 1% ME, Oysters and Mexicanas get 1.5% ME. If I grew cubes I'm guessing I would use 1.5% ME for them as well.

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OfflineHindsight
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: ruawakeyet] * 1
    #28170503 - 02/03/23 09:06 AM (1 year, 1 month ago)

Good pan mycelium should look like this:

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Offlineesotericbound
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: Hindsight] * 1
    #28170516 - 02/03/23 09:24 AM (1 year, 1 month ago)

Excellent, that certainly looks healthy, I'll keep trying.. Thanks for the input everyone


--------------------
"...If you have any kind of intellectual filter at all, you quickly can satisfy yourself that our best efforts are nothing more than half completed stories told around the campfire; we don't actually know what our predicament is."  -Terence McKenna

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OfflineAspectOfTheCreator
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: Hindsight]
    #28170812 - 02/03/23 01:51 PM (1 year, 1 month ago)

Is that a monoculture? Or is it multispore?


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OfflineHindsight
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: AspectOfTheCreator] * 1
    #28170854 - 02/03/23 02:24 PM (1 year, 1 month ago)

If you are asking me, it is MS. T1 transfer from spore germination plate.

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Offlineesotericbound
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: AspectOfTheCreator]
    #28170858 - 02/03/23 02:26 PM (1 year, 1 month ago)

I wish it was monoculture; this is multispore from print.  My normal method with cubes is MS->agar->clone healthy looking fruit from agar to new dish->grain...

This is my first time with pans and not sure if they'll fruit as easily on petri so I'll probably just work toward an isolate and spawn that to grain, worry about cloning later


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"...If you have any kind of intellectual filter at all, you quickly can satisfy yourself that our best efforts are nothing more than half completed stories told around the campfire; we don't actually know what our predicament is."  -Terence McKenna

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