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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, 11 months 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, 11 months 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, 11 months 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, 11 months 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, 11 months 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, 11 months 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, 11 months 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, 11 months 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, 11 months 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, 11 months 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, 11 months 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, 11 months 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


--------------------
"...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, 11 months 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, 11 months 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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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: esotericbound]
    #28170480 - 02/03/23 08:36 AM (1 year, 11 months 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, 11 months 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, 11 months ago)

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


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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: Hindsight]
    #28170812 - 02/03/23 01:51 PM (1 year, 11 months 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, 11 months 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, 11 months 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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Offlineesotericbound
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: Hindsight]
    #28170867 - 02/03/23 02:35 PM (1 year, 11 months ago)

Hindsight, on your agar plate that transfer piece is very round... is that something you take time to cut little circle out of dish it came from for some reason?  My transfers are always just little triangular wedges.


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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]
    #28171729 - 02/04/23 07:35 AM (1 year, 11 months ago)



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Offlineesotericbound
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: AspectOfTheCreator]
    #28171865 - 02/04/23 10:11 AM (1 year, 11 months ago)

:ooo: thanks for that - gorgeous agar work!


--------------------
"...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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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: esotericbound]
    #28172089 - 02/04/23 01:32 PM (1 year, 11 months ago)

That’s not pan mycelium. It is Ps. Mexicana var Jalisco. It even says so in writing on your plates. It looks like Mexicana mycelium too. You got confused about the genus that you inoculated.

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Offlineesotericbound
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: LookinOut] * 1
    #28172158 - 02/04/23 03:09 PM (1 year, 11 months ago)

No, "Panaeolus Cyanescens Jalisco Mexico" is the description on the website, the order confirmation, and receipt.


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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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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: LookinOut]
    #28172606 - 02/04/23 08:07 PM (1 year, 11 months ago)

Quote:

LookinOut said:
That’s not pan mycelium. It is Ps. Mexicana var Jalisco. It even says so in writing on your plates. It looks like Mexicana mycelium too. You got confused about the genus that you inoculated.




That was my first suspicion, and I made the same mistake with a member talking about Natal Pan Cyan. Once I searched it, I actually found a spore vendor selling Pan Cyan spores from Natal.

Same kind of thing with Pan Cyan Jalisco; it has even been crossed with other pans.

To me, unfortunately, it just looks like mold, but I'm not very experienced with Mexicana on agar.

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Offlineesotericbound
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: esotericbound] * 2
    #28173295 - 02/05/23 11:17 AM (1 year, 11 months ago)

T1 Pan plates look promising - all parameters the same except different agar recipe (1000mL H2O : 8g ME : 4g potato flakes : .4g peptone)






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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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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: esotericbound]
    #28173327 - 02/05/23 11:41 AM (1 year, 11 months ago)

Those are looking VERY promising!

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OfflineAspectOfTheCreator
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: esotericbound]
    #28173469 - 02/05/23 01:39 PM (1 year, 11 months ago)

Those look more like what I got.



Distinctive Pan myc

:thumbup:

I'd recommend atleast one more transfer before grains.


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Edited by AspectOfTheCreator (02/05/23 01:41 PM)

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Offlineesotericbound
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: AspectOfTheCreator] * 1
    #28173509 - 02/05/23 02:16 PM (1 year, 11 months ago)

What is reasoning for another transfer?  If it appears I don't have an isolate on these plates, I can understand need for another transfer.  I thought I read somewhere here that too much mycelium expansion (i.e. more transfers) decreases the tenacity of the mycelium and because of that, transfers are to be minimized, is this not correct?

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InvisibleBaba Yaga
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: esotericbound] * 1
    #28173527 - 02/05/23 02:29 PM (1 year, 11 months ago)

You can send this to grain if it still looks alright after it grew out a bit further. Just make sure you have a couple of plates as a backup you can go back to or take a transfer at the same you send this to grain, keep the rest of the plates around......something along these lines.

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OfflineAspectOfTheCreator
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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: esotericbound]
    #28173967 - 02/05/23 08:28 PM (1 year, 11 months ago)

It may or may not be necessary. I just like making atleast 2 transfers away from the last contaminated plate.


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Re: Panaeolus Cyanescens multispore growth on agar appearance? [Re: AspectOfTheCreator]
    #28174216 - 02/05/23 10:27 PM (1 year, 11 months ago)

Those first plates may have just been a funky agar recipe, it can make mycelium do some really strange things.

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