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Offlineinfectbilly
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PF Tek black horizontal line developed during later part of consolidation, help?
    #26798996 - 07/02/20 10:31 AM (3 years, 6 months ago)

Hello!

First time grower here, so excuse my ignorance, but I couldn't find info on this specific occurrence anywhere, so hoping you'll be able to help.

I've got 5 jars which were developing beautifully up until mid-consolidation week, when a thin black (almost) ring started appearing on all of the jars in the same place (see pic below, just below the vermiculite layer). I'm guessing it's not great, but just want to understand what it is and what could have lead it to happen so that I can prevent it next time.

The height seems similar to where the needle would have touched the glass, but the needle was removed from a sterile package and not touched before inoculating the first two jars and then was flame-sterilized before moving on to the other three..

Finally, does this mean I throw them all out or is there a way to save some of the progress? E.g. cut off the top of the cake?

In theory, I would have been dunking them in 2-3 days.


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OfflineTrichodingus
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Re: PF Tek black horizontal line developed during later part of consolidation, help? [Re: infectbilly]
    #26799391 - 07/02/20 01:38 PM (3 years, 6 months ago)

Hard to say why with the info provided.
Giving us a run down of how you prepared everything will help the community help you.

That cake is done man. That's sporulating mold in there. I personally wouldn't open that in my home as there are millions and millions of spores in there.


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Offlineinfectbilly
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Re: PF Tek black horizontal line developed during later part of consolidation, help? [Re: Trichodingus]
    #26800622 - 07/03/20 02:12 AM (3 years, 6 months ago)

Firstly, thank you for replying!

The process I went through:
1) prepped the substrate at a 1:1:2 (brf:water:verm) ratio
2) loaded the glasses (couldn't get hold of jars) with the substrate, filled top layer of vermiculite, put 2 pieces of foil over top of each glass with a rubber band holding them in place and pressure cooked at 12psi (the max my PC can go) for 2 hours, then letting it cool overnight in the PC
3) prepped the environment for inoculation by wiping the desk down with 99% isopropanol, set up a candle on the side for needle sterilization, set my jars and spore syringe on the side
4) removed the top foil layer and injected each jar at 4 points through the lower foil layer (each 90degrees apart), moving the needle below the verm layer on an angle until it touched the glass. 1.5ml each jar. Note that after the 2nd jar I flame sterilized the needle. While this could be a point of failure, I don't see why the first jar would show exactly the same contam pattern if so.
5) put the jars in a place with some light but out of direct sunlight and waiter. important note - I ended up needing to move the jars quite a few times, which I suspect could be a point of failure if that meant the verm layer moved enough to let contams through..
6) all 5 jars fully colonized ~8-10 days later and soon after started developing the black ring you see in the photo.

Logic dictates that all jars showing the same exact pattern of contamination means it's the environment rather than something done during inoculation, at least in my head. I just don't know what..

The one thing I'm not sure about because I found conflicting info online was whether I need to remove the cover once inoculated or not. What I did in the end was remove the foil entirely after day 3 or 4, which seemed to speed up the colonization. I did it as most people said the verm layer should act as a barrier, not the foil/cover.

While it's slightly disappointing these first jars are gone, I am definitely going at it again until I succeed, just want to find out as much as possible so that I don't repeat any mistakes..

EDIT: maybe worth adding to the whole "contam moved through the verm layer" idea - the cakes contracted significantly over colonization, to a point where pieces of verm were able to fall through between the cake and the glass. Not sure if that's normal? if not - what am i doing wrong?


Edited by infectbilly (07/03/20 05:19 AM)


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OnlineLogicaL ChaosM
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Re: PF Tek black horizontal line developed during later part of consolidation, help? [Re: infectbilly]
    #26800723 - 07/03/20 05:28 AM (3 years, 6 months ago)

Looks like bruizing to me, maybe from being too dry (no top layer of verm, open -top glass). If its Trich mold, the texture should look soft and powdery like velvet.

Can u get another pic but with a straight-on angle and closer to the line-band?


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Offlineinfectbilly
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Re: PF Tek black horizontal line developed during later part of consolidation, help? [Re: LogicaL Chaos]
    #26801247 - 07/03/20 10:49 AM (3 years, 6 months ago)

Now that you say it, it does look the same tint of blue as I've seen on bruised cubes.. The plot thickens! (thank you for introducing a sliver of hope btw)

I tried my best, but it's hard to capture when it's behind glass. I wouldn't describe it as you described Trich, it's more like it's part of the mycelium it seems, so bruising is an option!



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OfflineTrichodingus
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Re: PF Tek black horizontal line developed during later part of consolidation, help? [Re: infectbilly]
    #26805370 - 07/05/20 02:10 PM (3 years, 6 months ago)

Look at me being overzealous in throwing away your cakes 🤦

Keep us updated on how this turns out!


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