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Offlinemov369


Registered: 03/12/20
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Green Mold in Spawn Jars
    #26995060 - 10/20/20 05:50 PM (3 years, 3 months ago)

Hi,

I've recently moved from agar to spawn colonisation stage and face a problem with green mold in many of my jars (see pics).





I use organic rye berries (https://www.amazon.co.uk/gp/product/B0050BAPHC/ref=ppx_yo_dt_b_asin_title_o04_s00?ie=UTF8&psc=1), soaked for 24hours, boiled/simmered for 15 mins, left until the surface was dry and PC'ed for 90mins at 14psi (max for my PC, will sterilise for longer next time). Inoculated in a SAB with all sterile precautions using clean myc grown on petris.

My jars have a hole drilled in the cap stuffed with polyfill and are left out in the open on a shelf in the corner of my room close to the radiator (otherwise can't keep the temp at ~23C).

Can anyone give any suggestions as to why they developed the green mold? Would keeping the jars in a closed closet reduce the amount of contamination?

Also, I put quite a lot of myc agar wedges into my jars, so if they will grow through the spawn pretty fast and not let the green mold develop too much, would it be safe to then mix it with substrate and fruit it? Thanks a lot!


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OfflinePrincessRosalina
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Re: Green Mold in Spawn Jars [Re: mov369]
    #26995070 - 10/20/20 05:59 PM (3 years, 3 months ago)

Throw 'em in the bin and start more jars.


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Offlinemov369


Registered: 03/12/20
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Re: Green Mold in Spawn Jars [Re: PrincessRosalina]
    #26995081 - 10/20/20 06:04 PM (3 years, 3 months ago)

Thanks! Any ideas how to prevent that though and what I've done wrong? It's already a second time I encounter this issue, don't want to just waste time and grain :frown:


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Offlineedgar1337
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Re: Green Mold in Spawn Jars [Re: mov369]
    #26995092 - 10/20/20 06:08 PM (3 years, 3 months ago)

I'm too inexperienced to say, but for myself, most of the time I fail its due to too much moisture/water in the jars.

Then again, there are a million ways something can get contaminated unless you have some insane setup with a flowhood and completely sterile work space. According to BODs tek, the grains should be dry - the drier the better. You should check his tek: https://www.shroomery.org/forums/showflat.php/Number/24126032

It seems to have a high success rate. He also claims the overnight soak isn't necessary, just boiling and PC.


--------------------

"Mistakes are, after all, the foundations of truth, and if a man does not know what a thing is, it is at least an increase in knowledge if he knows what it is not."

-Carl Jung


"The current state of knowledge is a moment in history, changing just as rapidly as the state of knowledge in the past has ever changed and, in many instances, more rapidly."
-Jean Piaget


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Offlinemov369


Registered: 03/12/20
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Re: Green Mold in Spawn Jars [Re: edgar1337]
    #26995108 - 10/20/20 06:21 PM (3 years, 3 months ago)

Cheers, I'll try this out next time!


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OfflineNichrome
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Re: Green Mold in Spawn Jars [Re: mov369] * 1
    #26995237 - 10/20/20 07:27 PM (3 years, 3 months ago)

Get some real SFD's synthetic filter discs or CFD's cellulose filter discs. 90 minutes is fine some people regularly do less. Do you use an SAB or a laminar flow hepa filter? If so the mold is either coming off of you at the time of inoculation or it is in your inoculant.


--------------------
Better to be deprived of food for three days, than tea for one.


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Edited by Nichrome (10/20/20 09:59 PM)


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Offlinemeowjinx
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Re: Green Mold in Spawn Jars [Re: mov369]
    #26995246 - 10/20/20 07:31 PM (3 years, 3 months ago)

Quote:

mov369 said:
Thanks! Any ideas how to prevent that though and what I've done wrong? It's already a second time I encounter this issue, don't want to just waste time and grain :frown:




Trich (green mold) is the bane of my existence and the reason for a lot of my misery

Using lots of pieces of agar is the opposite of what I would do in your situation. You cannot outrun Trich, it will pop up at any stage, including spawning and fruiting

It's hard to spot on agar because it's mycelium starts off white, just like cubes. It doesn't turn green until it sporulates, which is who knows when the fuck (when it's too late is the point). So you're better off using smaller pieces of agar so that you are less likely to transfer some Trich myc along with the cubes to your grain jars

Toss anything and everything that goes green. Do not bother trying to save it. At least that's my opinion


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OfflineNichrome
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Re: Green Mold in Spawn Jars [Re: meowjinx]
    #26995498 - 10/20/20 10:20 PM (3 years, 3 months ago)

I've cleaned up lots of cultures and trich aint no trick. Just transfer clean mycelium away to new plates. Often you can eliminate the trich with one good transfer. The mold looks different from the mycelium you want and it's not that hard to differentiate. I've yet to see trichoderma "run" alongside or with cube mycelium. It almost always pops up away from the mycelium in the plate and one will almost always overtake the other eventually and sometimes after epic battles where much metabolite is produced. Trich especially shows green (sporulates) on agar pretty fast comparatively to other molds. I grow mold on purpose (aspergillus, rhizopus, penicillium, and yes even trichoderma) regularly and it is nothing to fear or worry about and is easily eliminated from the situation.

It may take some practice but if you buy a whole box of dishes they're around $0.18 usd per plate. Agar and malt are cheap. You can, and many people do, pour plates in an SAB.  It just takes some practice. If it is not working then you are doing it wrong and you should do more research, practice, and reach out to the community till it does. No need to continue to struggle. No need to give up. No need to ever "loose" a culture unless you're ready to let it go.

Just for shits and giggles here is a photo of Aspergillus Oryzae spores sprouting in a water droplet suspended within the colony.



The stuff in your jar looks more like penicillium than trich but it'd be really hard to tell without a microscope. I see lots of molds misidentified as Trichoderma on this forum but it's not too important as mold on your shit is mold on your shit and is never good.

Good luck and keep practicing.


--------------------
Better to be deprived of food for three days, than tea for one.


Freedom is not the right to do as you please, but the liberty to do as you should. ~Emerson


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Offlinemov369


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Re: Green Mold in Spawn Jars [Re: Nichrome]
    #26995753 - 10/21/20 05:19 AM (3 years, 3 months ago)

I've managed to isolate clean myc on petri dishes, at least from what I can tell.



My problem with mold is at the stage of spawn colonisation.

From what I've gathered I'll:

1. Get synthetic disc filters to aid polyfill.
2. Make sure the insides of the jars and grains are dry before inoculating.
3. Be more careful while inoculating.
4. Clean up the space where I store the jars and in the future monotubs with iso & bleach (walls / floor/ spray air with water, let everything sit then carefully mop and cover that space with shower curtain? Woudl that be a good idea? I don't have space to create a complete clean room, but I'll try to make a clean corner that is isolated by a curtain from the rest of the room which I try to keep clean anyways).


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OfflinePearl
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Re: Green Mold in Spawn Jars [Re: Nichrome] * 2
    #26995754 - 10/21/20 05:20 AM (3 years, 3 months ago)

Have you tried keeping a control jar (one that you prep the same way but don't inoculate) to see if it also fails?

Are you venting your PC for 10mins before you start your timer? If not, do so AND PC for 2hrs then see how your control does alongside any inoculated jars.

If your control is fine and the inoculated jars fail, it's your culture.

If your control also fails, it's your prep and MAYBE your culture.

If none fail, you're laughing.


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OfflinePearl
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Re: Green Mold in Spawn Jars [Re: mov369]
    #26995757 - 10/21/20 05:23 AM (3 years, 3 months ago)

Quote:

mov369 said:
1. Get synthetic disc filters to aid polyfill.
2. Make sure the insides of the jars and grains are dry before inoculating.
3. Be more careful while inoculating.
4. Clean up the space where I store the jars and in the future monotubs with iso & bleach (walls / floor/ spray air with water, let everything sit then carefully mop and cover that space with shower curtain? Woudl that be a good idea? I don't have space to create a complete clean room, but I'll try to make a clean corner that is isolated by a curtain from the rest of the room which I try to keep clean anyways).




1. Yep, will help.
2. Dry before PC'ing. If you're not already doing so, do the toilet paper test.
3. Never a bad thing
4. Also never a bad thing. But if you're prepped right and you have good filters, not 100% needed. But again, being clean is never bad. :wink:



You plate looks good so I gotta say the weak link is in prep...


Edited by Pearl (10/21/20 05:24 AM)


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Offlinemov369


Registered: 03/12/20
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Re: Green Mold in Spawn Jars [Re: Pearl]
    #26995758 - 10/21/20 05:24 AM (3 years, 3 months ago)

That's a good idea! Will do.

What do you mean by venting the PC? I put jars into the PC, close and start the gas, wait until it reaches 14 psi (max for my PC) and then start the timer. I will however do a bit longer next time, maybe 1h 45m - 2h.


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OfflinePearl
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Re: Green Mold in Spawn Jars [Re: mov369]
    #26995759 - 10/21/20 05:28 AM (3 years, 3 months ago)

Essentially:
Put the PC on high heat
Wait for a steady stream of steam to be constantly being expelled from the vent
Set timer for 10 mins
When timer is up, put on weight to bring to pressure
Set time for 90mins to 2hrs

(don't let it boil dry)


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Offlinemov369


Registered: 03/12/20
Posts: 25
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Re: Green Mold in Spawn Jars [Re: Pearl]
    #26995770 - 10/21/20 05:41 AM (3 years, 3 months ago)

Quote:

Pearl said:
2. Dry before PC'ing. If you're not already doing so, do the toilet paper test.





Yeah that's what I meant, my bad :laugh:

Are these coffee making machine discs any good? They do fit the caps of my jars diameter-wise, should I just stick them in on the inside before PC-ing?

https://www.amazon.co.uk/AeroPress-TKC81R24-Filter-Papers-White/dp/B000LTOCSG/ref=sr_1_14?dchild=1&keywords=synthetic+air+filters&qid=1603279661&sr=8-14&th=1

Otherwise I found these ones on a UK mushroom shop, but they're breaking my bank for sure :frown:

https://www.cylocybe.com/product/synthetic-filter-discs-sfds/

Quote:

Pearl said:
Essentially:
Put the PC on high heat
Wait for a steady stream of steam to be constantly being expelled from the vent
Set timer for 10 mins
When timer is up, put on weight to bring to pressure
Set time for 90mins to 2hrs

(don't let it boil dry)




Thanks! Mine doesn't have weight on it, just psi modes, one of which is 14 psi. I just wait until it reaches 14 psi and then start the timer.


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OfflinePBJ710
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Re: Green Mold in Spawn Jars [Re: mov369]
    #26995862 - 10/21/20 07:38 AM (3 years, 3 months ago)

Definitely vent your PC - leaving air in there can cause what you are seeing.  Your jar GE filtration is definitely very suspect as well.  Those paper filters will not make it through a PC cycle - use a real hydrophobic SFD.  They are worth it and a 1 time purchase.  You can use the cheaper 29mm ones or cut up the full size ones to cover a 3/8"-1/2" hole in the lid.



Edited by PBJ710 (10/21/20 07:44 AM)


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Offlinemov369


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Re: Green Mold in Spawn Jars [Re: PBJ710]
    #26995941 - 10/21/20 08:45 AM (3 years, 3 months ago)

Sorry for a stupid question, but do I vent with the jars in the PC or should I put them in after I vented?


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OfflineOne of Us
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Re: Green Mold in Spawn Jars [Re: mov369]
    #26995950 - 10/21/20 08:51 AM (3 years, 3 months ago)

Definitely with the jars in the PC. The reason for venting is you want to purge all the air that is in the jars and replace it with steam. Steam will get hotter and sterilize better than air.


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OfflineNichrome
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Re: Green Mold in Spawn Jars [Re: One of Us]
    #26996138 - 10/21/20 10:48 AM (3 years, 3 months ago)

The PC will actually reach full pressure faster with propper venting too.


--------------------
Better to be deprived of food for three days, than tea for one.


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OfflineBig_Dub
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Re: Green Mold in Spawn Jars [Re: Nichrome]
    #26996148 - 10/21/20 10:51 AM (3 years, 3 months ago)

yeah venting PC is necessary, they even say this in the PC manual.

The best way to stop molds is to get better cultures on agar. just keep practicing. make lots of petri plates and be really mindful of how you work in SAB. contamination will happen from time to time, but you can greatly reduce it by getting good technique


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Offlinetiptrippy
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Re: Green Mold in Spawn Jars [Re: mov369]
    #26996151 - 10/21/20 10:53 AM (3 years, 3 months ago)

Quote:

mov369 said:
Quote:

Pearl said:
2. Dry before PC'ing. If you're not already doing so, do the toilet paper test.





Yeah that's what I meant, my bad :laugh:

Are these coffee making machine discs any good? They do fit the caps of my jars diameter-wise, should I just stick them in on the inside before PC-ing?

https://www.amazon.co.uk/AeroPress-TKC81R24-Filter-Papers-White/dp/B000LTOCSG/ref=sr_1_14?dchild=1&keywords=synthetic+air+filters&qid=1603279661&sr=8-14&th=1

Otherwise I found these ones on a UK mushroom shop, but they're breaking my bank for sure :frown:

https://www.cylocybe.com/product/synthetic-filter-discs-sfds/

Quote:

Pearl said:
Essentially:
Put the PC on high heat
Wait for a steady stream of steam to be constantly being expelled from the vent
Set timer for 10 mins
When timer is up, put on weight to bring to pressure
Set time for 90mins to 2hrs

(don't let it boil dry)




Thanks! Mine doesn't have weight on it, just psi modes, one of which is 14 psi. I just wait until it reaches 14 psi and then start the timer.




Are you using a digital pressure cooker like an instant pot? There may not be a way to vent that specific PC.


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