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Offlinesmokedtrolli
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9/9 oat jars contam'd with what appears to be Penicillium. Why?
    #26431456 - 01/13/20 02:20 PM (4 years, 2 months ago)

I inoculated these jars with agar wedges that were made about a month ago. The plate I used for these showed very strong rhizomorphic growth with complete uniformity. There was no suspicion of other kinds of growth on the plate that wasn't mycelium.

I prepped the oats by boiling for ~40 minutes, strained them in a colander, and then laying them out on paper towels to dry them as much as possible before adding them to the jars and PCing. I PCed at 15 PSI for 90 minutes and didn't touch them until the following day.

When I removed them, there were no signs of wetness. I inoculated using a SAB, and x-acto knife that was heated via butane torch in between each jar. I wore gloves (though, my forearms were exposed) and tried to avoid moving anything that was not sterile above the jar openings or the agar plate during the process. As I was dropping the wedge into the jars, I did my best to only remove the jar's lid for as long as it took me to get it off the knife.

Afterward, the jars were kept in my basement at low temps (60s I'm assuming). I waited for some mycelium to grow off onto local grains surrounding the wedges before I shook them. After the shake is when the same kind of mold appeared in every jar. Bluish-green, somewhat fuzzy. I'm 99.99% sure it is not bruising.

What are the theories here? Did the plate have covert growth? Did the grains pick up something from the paper towels/colanders? Is my sterile technique dogshit? Or just absolutely horrendous luck?


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InvisibleSpunkyMonkey88
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Re: 9/9 oat jars contam'd with what appears to be Penicillium. Why? [Re: smokedtrolli]
    #26431464 - 01/13/20 02:25 PM (4 years, 2 months ago)

It could be any number of things... improper sterilization of grains, sloppy SAB work or dirty plates... if they are all contaminated I'd probably lean towards a dirty plate...


U got any pics?

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Offlinesmokedtrolli
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Re: 9/9 oat jars contam'd with what appears to be Penicillium. Why? [Re: SpunkyMonkey88]
    #26431470 - 01/13/20 02:28 PM (4 years, 2 months ago)

Yeah, here are pics:







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InvisibleSpunkyMonkey88
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Re: 9/9 oat jars contam'd with what appears to be Penicillium. Why? [Re: smokedtrolli]
    #26431477 - 01/13/20 02:32 PM (4 years, 2 months ago)

That spawn is contaminated for sure but do u have any pics of your agar

Edited by SpunkyMonkey88 (01/13/20 02:33 PM)

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Offlinesmokedtrolli
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Re: 9/9 oat jars contam'd with what appears to be Penicillium. Why? [Re: SpunkyMonkey88]
    #26431479 - 01/13/20 02:32 PM (4 years, 2 months ago)

Quote:

SpunkyMonkey88 said:
That spawn is contaminated




As in, the plate was?


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InvisibleSpunkyMonkey88
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Re: 9/9 oat jars contam'd with what appears to be Penicillium. Why? [Re: smokedtrolli]
    #26431482 - 01/13/20 02:34 PM (4 years, 2 months ago)

Quote:

smokedtrolli said:
Quote:

SpunkyMonkey88 said:
That spawn is contaminated




As in, the plate was?



I would think so I dont see any healthy mushroom mycelium I'm thinking you inoculated your jars with mold

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Re: 9/9 oat jars contam'd with what appears to be Penicillium. Why? [Re: SpunkyMonkey88]
    #26431485 - 01/13/20 02:35 PM (4 years, 2 months ago)

Either that or you didnt PC your jars long enough, most people go two hours...

I would think 100% moldy jars either means you inoculated with mold or your grains wernt sterile. I'm thinking your plate wasn't even mushrooms mycelium itd help if u had some pics

Edited by SpunkyMonkey88 (01/13/20 02:38 PM)

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Invisiblerickyswamps
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Re: 9/9 oat jars contam'd with what appears to be Penicillium. Why? [Re: SpunkyMonkey88]
    #26431509 - 01/13/20 02:43 PM (4 years, 2 months ago)

PC cycles always kill mold.  Its probably from the plates if it was 9 out of 9.  If they look perfect, then it is from SAB technique.

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Offlinesmokedtrolli
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Re: 9/9 oat jars contam'd with what appears to be Penicillium. Why? [Re: SpunkyMonkey88]
    #26431513 - 01/13/20 02:44 PM (4 years, 2 months ago)

Quote:

SpunkyMonkey88 said:
Either that or you didnt PC your jars long enough, most people go two hours...

I would think 100% moldy jars either means you inoculated with mold or your grains wernt sterile. I'm thinking your plate wasn't even mushrooms mycelium itd help if u had some pics





This is what's leftover. The middle of it contains a piece from a previous plate.



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Edited by smokedtrolli (01/13/20 02:59 PM)

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InvisiblebodhisattaMDiscordReddit
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Re: 9/9 oat jars contam'd with what appears to be Penicillium. Why? [Re: smokedtrolli]
    #26431580 - 01/13/20 03:13 PM (4 years, 2 months ago)

Your two options are you didn't notice your plate wasn't 100% healthy.
Or it got in during the transfer/inoculation

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Offlinesmokedtrolli
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Re: 9/9 oat jars contam'd with what appears to be Penicillium. Why? [Re: bodhisatta]
    #26431607 - 01/13/20 03:23 PM (4 years, 2 months ago)

Quote:

bodhisatta said:
Your two options are you didn't notice your plate wasn't 100% healthy.
Or it got in during the transfer/inoculation




That's what I figured. Probably during transfer if I had to guess.


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InvisiblebodhisattaMDiscordReddit
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Re: 9/9 oat jars contam'd with what appears to be Penicillium. Why? [Re: smokedtrolli]
    #26431608 - 01/13/20 03:23 PM (4 years, 2 months ago)

Or your gas exchange filters are insufficient or incorrectly made

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Re: 9/9 oat jars contam'd with what appears to be Penicillium. Why? [Re: bodhisatta]
    #26431663 - 01/13/20 03:47 PM (4 years, 2 months ago)

Quote:

bodhisatta said:
Or your gas exchange filters are insufficient or incorrectly made




Ya those are possibilities I didnt think of... It doesnt look like whatever is in your jars was on your plate but who knows they are way overgrown but look good to me....


I'm gonna say you didnt PC properly after seeing ur leftover plate...but it's all just an educated guess at this point...

You'd have to really have some shitty technique in the SAB to introduce contamination 9 out of 9 times in between the plate and the jar(s)

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Offlinesmokedtrolli
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Re: 9/9 oat jars contam'd with what appears to be Penicillium. Why? [Re: bodhisatta]
    #26431673 - 01/13/20 03:49 PM (4 years, 2 months ago)

Quote:

bodhisatta said:
Or your gas exchange filters are insufficient or incorrectly made




On the jars? They're 1/4in. holes stuffed with polyfill. Is that alright or should I use tyvek filters instead?


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Re: 9/9 oat jars contam'd with what appears to be Penicillium. Why? [Re: smokedtrolli]
    #26431675 - 01/13/20 03:50 PM (4 years, 2 months ago)

Poly is fine. If it wasn't stuffed tight enough that could be your problem though

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Offlinesmokedtrolli
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Re: 9/9 oat jars contam'd with what appears to be Penicillium. Why? [Re: SpunkyMonkey88]
    #26431676 - 01/13/20 03:51 PM (4 years, 2 months ago)

I mean, I suppose I could have fucked up PCing, but that's so incredibly simple I don't see how anyone could fuck it up unless they were way below PSI or misjudged time by an hour.


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Offlinesmokedtrolli
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Re: 9/9 oat jars contam'd with what appears to be Penicillium. Why? [Re: bodhisatta]
    #26431682 - 01/13/20 03:53 PM (4 years, 2 months ago)

Hmm... I didn't quite grab the biggest ball of polyfill when I made those lids... I suppose it could have been somewhat loose.


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Re: 9/9 oat jars contam'd with what appears to be Penicillium. Why? [Re: smokedtrolli]
    #26431688 - 01/13/20 03:59 PM (4 years, 2 months ago)

Well if it was from the agar/lid/inoculation it would have started from the top of the jar... if it were the grains itd be sporadic throughout the entire jar...

Where did you first see the contamination take hold? Any kind of pattern? That may help you figure out the origin

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Re: 9/9 oat jars contam'd with what appears to be Penicillium. Why? [Re: SpunkyMonkey88]
    #26431691 - 01/13/20 04:01 PM (4 years, 2 months ago)

That is assuming you didnt shake your jar after dropping the agar...

I always like to drop it and let it sit for a while to see what developers before shaking it that way you know what is going on and what you're spreading VIA shaking...

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Offlinesmokedtrolli
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Re: 9/9 oat jars contam'd with what appears to be Penicillium. Why? [Re: SpunkyMonkey88]
    #26431693 - 01/13/20 04:03 PM (4 years, 2 months ago)

I shook most of them after not seeing any mold and having some mycelium growth off of the wedge. Of the ones I did not shake, the contamination actually did originate from the top.


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