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Offlineaireair
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Registered: 02/28/24
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Huge contamination issue
    #28679599 - 02/28/24 12:10 PM (10 months, 13 days ago)

Hello all.

So I'm having some insane trouble with this and wanted to reach out to see if anyone had any idea of what I maybe doing wrong.

I've been through probably 500-600 jars with 100% contam rate.

I tried basically every tek online. Multiple runs of each. Using 4x2 brand new flow hood. Tried using oats, millet and popcorn. All my agar work is clean but somehow every jar ends up smelling sour once colonization starts.


Mainstay lids with micropose filter patches.

I even tried to buy sterilized grain bags with LC to inoculate.. still same exact smell in all 10 bags.

Temperature between 60 and 70.

stared this hobby up to keep busy and have some fun but thus far its been totally frustrating lol.


I've asked multiple different places and nobody seems to have any idea what I could be doing wrong.

Has anyone else experienced this?


I've been trying for around a year and a half to dial into perfect conditions, spent probably 100 hours reading, but it's been fail after fail. Starting to lose hope.

Hundreds of perfectly clean plates. Following every tek to the T and still having issues I really don't get what's happening here.

Anyone with ideas I am all ears.

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Invisiblejoze
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Re: Huge contamination issue [Re: aireair]
    #28679775 - 02/28/24 02:17 PM (10 months, 13 days ago)

This should be in the mushroom cultivation board.

If your agar work is clean, but your jars are turning sour, then there are two likely scenarios. Either 1) your grains are not properly sterilized or 2) you are somehow causing contamination while inoculating your grains.

As detailed as possible, please describe your methods for both sterilizing your grains and for inoculating them.


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OfflineNeurotech
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Re: Huge contamination issue [Re: aireair]
    #28679826 - 02/28/24 02:52 PM (10 months, 13 days ago)

Can you elaborate on the signs of contamination? Wondering if just a sour smell is the only thing? No visual contamination? Pics?

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OfflineNorthernerM
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Re: Huge contamination issue (moved) [Re: aireair]
    #28679844 - 02/28/24 03:01 PM (10 months, 13 days ago)

This thread was moved from The Psychedelic Experience.

Reason:
Goes here

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OfflinePuffM4gicDr4gon
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Re: Huge contamination issue (moved) [Re: Northerner]
    #28680038 - 02/28/24 04:56 PM (10 months, 13 days ago)

I had a wet rot problem similar to you. It was fixed by upping my PC time to 4 hours and properly venting my PC before starting the timer. 4 hours is probably overkill, but it works so I haven't bothered to reduce it


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Promoted from Captain to Dragon

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Offline99.99
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Re: Huge contamination issue (moved) [Re: PuffM4gicDr4gon]
    #28680150 - 02/28/24 06:02 PM (10 months, 13 days ago)

Is your flow hood meeting specification? Does it filter 99.99% for sure does it say so on the stickers that’s on the flow hood

You need to run your flow hood for an hour or so in a enclosed environment like a room to get things sterile enough

You need to vent your pressure cooker for 10 minutes before you cook things it needs to be at least 15 psi for at least two hours

You need to remove your jars from the pressure cooker in front of the flow hood and let them cool in front of the flow hood

You need to wear rubber gloves, anytime you’re touching anything and I would strongly recommend spraying everything down before the work starts with a 10% bleach to water solution

We need to know what sterile agar plates look like, and understand what mycelium looks like

Or whatever you buy needs to come from a sponsor on this forum

Jars really suck ass I would strongly recommend using bags. They are extremely user-friendly for me.
All I do is 2 cups wheat and 2 cups water inside a bag … no stupid ass prep shit
roll up the top and close it together tightly with a binder clip

[50 Pack] Mushroom Growing Bags... https://www.amazon.com/dp/B07Q8VZ8X6?ref=ppx_pop_mob_ap_share

Seal them up like this before you cook them



And if you’re still having problems, you’re seriously fucked you need to pay somebody to come in person and see what the fuck you’re doing and set you straight

Edited by 99.99 (02/28/24 06:09 PM)

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OfflineOldManRiver
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Re: Huge contamination issue (moved) [Re: 99.99]
    #28680185 - 02/28/24 06:34 PM (10 months, 13 days ago)

Quote:

99.99 said:
Is your flow hood meeting specification? Does it filter 99.99% for sure does it say so on the stickers that’s on the flow hood

You need to run your flow hood for an hour or so in a enclosed environment like a room to get things sterile enough





My bet is that the flow hood situation is the issue, as well.  I don't think running for an extended period is necessary. If the hood is good, it should be good instantly.  I turn mine on and get to work immediately without any problems. 

OP, turn on your hood and open an agar dish in front of it for a minute, then monitor the dish.  That will tell you what you need to know.  If your hood was inexpensive, it may be bunk.  Just the filter for a good hood is $150+.

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Offline99.99
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Re: Huge contamination issue (moved) [Re: OldManRiver]
    #28680200 - 02/28/24 06:41 PM (10 months, 13 days ago)

If you knew what the problem was, you wouldn’t be having contamination issues, so why don’t you do like everybody else does and run the flow hood for a while

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Invisiblefungusul
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Registered: 07/16/20
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Re: Huge contamination issue (moved) [Re: 99.99]
    #28680249 - 02/28/24 07:16 PM (10 months, 13 days ago)

Welcome to Shroomery! :mushroom2:

Have you try to keep control jars and control agar plates?

You could keep a control jar or control agar plate not inoculated and check them after a couple of days. If they are contaminated, then you know what step in your process fails.

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OfflineSkropi
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Registered: 11/01/23
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Re: Huge contamination issue (moved) [Re: fungusul]
    #28680288 - 02/28/24 07:40 PM (10 months, 13 days ago)

I am 99% sure that the issue is the flow hood. Still, it can be anything. I would trust a SAB more than an inexpensive flow hood though. The advice to use a petri dish as control is perfect, and it will show you if this is true in a few days. Just use a petri dish that is definitely clean, I wouldn't use one that was poured less than 15-20 days before, temperature depending too of course. Meanwhile, do make a small batch in a SAB too.


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Ἄνδρα μοι ἔννεπε, Μοῦσα, πολύτροπον

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Offline99.99
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Re: Huge contamination issue (moved) [Re: Skropi]
    #28680292 - 02/28/24 07:45 PM (10 months, 13 days ago)

Quote:

Skropi said:
I am 99% sure that the issue is the flow hood. Still, it can be anything. I would trust a SAB more than an inexpensive flow hood though. The advice to use a petri dish as control is perfect, and it will show you if this is true in a few days. Just use a petri dish that is definitely clean, I wouldn't use one that was poured less than 15-20 days before, temperature depending too of course. Meanwhile, do make a small batch in a SAB too.




That’s only if he has a sterile, petri dish to begin with… if he can’t make clean spawn, how is he gonna make  a clean petri dish?

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OfflineSkropi
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Re: Huge contamination issue (moved) [Re: 99.99]
    #28680295 - 02/28/24 07:49 PM (10 months, 13 days ago)

You are right, maybe I wasn't clear, but he should use a dish that isn't inoculated, and see if something starts growing, to be perfectly sure 😁


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Ἄνδρα μοι ἔννεπε, Μοῦσα, πολύτροπον

Edited by Skropi (02/28/24 07:50 PM)

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InvisibleLadysKnight
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Re: Huge contamination issue (moved) [Re: Skropi] * 1
    #28680299 - 02/28/24 07:53 PM (10 months, 13 days ago)

Make no-pour agar plates. Test in flow and sab.

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Offline99.99
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Re: Huge contamination issue (moved) [Re: LadysKnight]
    #28680305 - 02/28/24 07:58 PM (10 months, 13 days ago)

Quote:

LadysKnight said:
Make no-pour agar plates. Test in flow and sab.




If his no poor Agar plates are not being sterilized well enough in the pressure cooker it won’t help either

You can’t just think it’s one thing and try to fix one thing you need to go in and refix everything to the correct way of doing it

He has tried that and wasted over 500 jars trying to figure out what the problem is

I had the same problem when I first started the slime bacteria mold that is always present on our bodies is very hard to distinguish on plates and you can’t see it in jars until it’s way too late

The only thing that has stopped that slime crap for me is using a 10% bleach solution
So I would suggest starting from step one using the bleach solution. I know a lot of people don’t like bleach, but it kills a lot more than other things do

Edited by 99.99 (02/28/24 08:16 PM)

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InvisibleLadysKnight
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Re: Huge contamination issue (moved) [Re: 99.99] * 2
    #28680315 - 02/28/24 08:14 PM (10 months, 13 days ago)

Seems agar is easier to sterilize than grain. I think op could pull off clean no-pour plates. Then wrap 2 immediately which if they all go bad, points toward the PC. Two more open in the flow hood for a minute, wrap. If they fail it's the hood. 2 more try a couple dry xfers, wrap, see what develops. Could try the same in a sab.
I think they can narrow things down with a run of no-pour plates.

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Offline99.99
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Re: Huge contamination issue (moved) [Re: LadysKnight]
    #28680319 - 02/28/24 08:19 PM (10 months, 13 days ago)

Quote:

LadysKnight said:
Seems agar is easier to sterilize than grain. I think op could pull off clean no-pour plates. Then wrap 2 immediately which if they all go bad, points toward the PC. Two more open in the flow hood for a minute, wrap. If they fail it's the hood. 2 more try a couple dry xfers, wrap, see what develops. Could try the same in a sab.
I think they can narrow things down with a run of no-pour plates.




I agree with what you are saying, but he does not say if he uses a Presto pressure cooker does not say how long he cooks things,  he doesn’t say what PSI nothing like that

He does say he can make clean agar plates, but that slime mold is very hard to see and pick out

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Offline99.99
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Re: Huge contamination issue (moved) [Re: 99.99]
    #28680326 - 02/28/24 08:23 PM (10 months, 13 days ago)

One more thing sterilize your tools with whatever you are pressure cooking don’t rely on flame sterilization

Wrap them up in aluminum foil, so they stay stay sterile until you open them in front of your flow hood

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InvisibleMycolorado
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Re: Huge contamination issue (moved) [Re: 99.99] * 1
    #28680347 - 02/28/24 08:44 PM (10 months, 13 days ago)

Using bleach and not trusting flame sterilization of tools is bonkers. I think we need a tad more info from op before we can truly parse this out.

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Offline99.99
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Re: Huge contamination issue (moved) [Re: Mycolorado]
    #28680357 - 02/28/24 08:51 PM (10 months, 13 days ago)

You cant wrap up bleach and flame sterilization all together in the same sentence

Flame sterilization of tools is great, but you can also miss spots
It would be a lot better to sterilize whatever you’re going to use in pressure cooker at 15 psi for half an hour ….then everything you touch and use is sterile. Not just hoping the blade is good enough on the scalpel
He doesn’t know 100% every step that he does is perfect, so why not start from the very beginning of the whole process and be 100% on everything that you do

Edited by 99.99 (02/28/24 08:56 PM)

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InvisibleMycolorado
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Re: Huge contamination issue (moved) [Re: 99.99] * 1
    #28680369 - 02/28/24 08:57 PM (10 months, 13 days ago)

It’s not necessary.  My tools sit on the bench in front of my hood for weeks.  There are likely some technique/equipment issues at hand here.  That could include improper flaming of tools but I’m guessing that’s pretty low on the issue list.

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