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InvisibleLadysKnight
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Registered: 10/09/15
Posts: 2,331
Re: Diagnosing contamination issues/reviewing technique [Re: phenyl] * 2
    #28205018 - 02/26/23 07:17 AM (1 year, 10 months ago)

Quote:

phenyl said:
I don't see anything mentioned about reasonably ruling out cultures contaminated with enmeshed bacteria or fungi. Very annoying and often subtle contamination vector.




:whathesaid:

Got plate pics?


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OfflineMushyMom
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Re: Diagnosing contamination issues/reviewing technique [Re: LadysKnight]
    #28239959 - 03/21/23 04:27 PM (1 year, 9 months ago)

Prepped grain jars - using WBS - added to boiling water, took off heat and let soak for 90 minutes.  Drained.  Loaded jars.  PC’d in jars with unmodified plastic lids (vented for 15 minutes then PC for 90).

3-3 Used LC from Shroomery sponsor.  Applied to agar at the same time as the jars to test. Plates were clean. 

3-16 - shook jars

These pictures were taken last night





I’m not nuts about the way the grain looks.  I’m in wait and see mode on these.

Concerns are
1. Old WBS
2. Unmodified lids - some are warping.

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OfflineMushyMom
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Re: Diagnosing contamination issues/reviewing technique [Re: MushyMom]
    #28239965 - 03/21/23 04:31 PM (1 year, 9 months ago)

Today - currently running jars using new Millet. 

Washed grains and drained.  Added grains to boiling water.  Removed from heat, covered and let sit for an hour.  Drained. Loaded jars.  Currently running with modified lids.  Planning to run for 2 hours.

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OfflineScrewup
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Re: Diagnosing contamination issues/reviewing technique [Re: MushyMom]
    #28239967 - 03/21/23 04:31 PM (1 year, 9 months ago)

Where’s the plate pics?


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OfflineHappinessStan
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Re: Diagnosing contamination issues/reviewing technique [Re: Screwup] * 1
    #28239997 - 03/21/23 04:52 PM (1 year, 9 months ago)

First, plate pics would really help, second, what contams are you facing? Mold and bacteria can happen for different reasons. Also, have you considered the filters on the jars maybe wicking contams in? Also, what does the lc look like. You're sure it's lc and not a spore syringe? These are the things I would address first.
I wouldn't shoot the bought lc to grain, I'd take some clean myc from the agar, transfer out a few times, then inoculate with that, either with a wedge or make it into an lc or li to inoculate.
That being said, nothing looks bad about those wbs jars to my eye. Especially after a shake.
Hope you work it out.:mushroom2:


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OfflineMushyMom
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Re: Diagnosing contamination issues/reviewing technique [Re: HappinessStan]
    #28240063 - 03/21/23 05:37 PM (1 year, 9 months ago)

Quote:

HappinessStan said:
First, plate pics would really help, second, what contams are you facing? Mold and bacteria can happen for different reasons. Also, have you considered the filters on the jars maybe wicking contams in? Also, what does the lc look like. You're sure it's lc and not a spore syringe? These are the things I would address first.
I wouldn't shoot the bought lc to grain, I'd take some clean myc from the agar, transfer out a few times, then inoculate with that, either with a wedge or make it into an lc or li to inoculate.
That being said, nothing looks bad about those wbs jars to my eye. Especially after a shake.
Hope you work it out.:mushroom2:




1. I had nice grows last year until I didn’t. We’re talking explosive green bins.
2. Yes it’s LC from a shroomery vendor. I know for sure it’s not spores.  But I tested it on agar when I noc’d the jars up and figured it it was shit I would just toss the jars
3. I’ve been working on my agar technique separate so I figure using the LC would help me separate my testing. I just put agar to grain last night too. Not my best agar but I had jars ready to go and figured why not. I need to load in a plate picture.
4. I’ve been running unmodified plastic lids. I am not nuts about them - it seems like I’m putting a little too much faith in the threads of the lid being sealed enough to keep out contam but open enough for air lol. 
5. My concerns with these jars - I’m seeing wet grains pressed against the jar on some. I see condensation on the top of some. And there’s one where the grains are looking red and pink. I’ve had worse jars and some of these look ok but some are questionable for me. I am going off of Stipes clean spawn checklist.

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Offlinetholos
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Re: Diagnosing contamination issues/reviewing technique [Re: MushyMom]
    #28240135 - 03/21/23 06:26 PM (1 year, 9 months ago)

Have you considered increasing your sterilization time in the PC? I've seen many people say they do 2-2.5 hrs for grain as a fail safe. Of course this would only help if the bacteria was present when you put your LC to grain

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OfflineMushyMom
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Re: Diagnosing contamination issues/reviewing technique [Re: MushyMom]
    #28240424 - 03/21/23 08:37 PM (1 year, 9 months ago)

These are my plates that I sent to grain last might. Nothing exciting to see yet. They looked like they were about to pin and I had grain I had to use so I figured why not. 





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OfflineMushyMom
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Re: Diagnosing contamination issues/reviewing technique [Re: tholos]
    #28240614 - 03/21/23 10:37 PM (1 year, 9 months ago)

Quote:

tholos said:
Have you considered increasing your sterilization time in the PC? I've seen many people say they do 2-2.5 hrs for grain as a fail safe. Of course this would only help if the bacteria was present when you put your LC to grain



So last year I kept upping the time and things kept getting worse. Like first I lost a tub or two. Then I lost a couple tubs. And then I was losing everything. So I don’t know if it made a difference or if everything was too far gone by that point :smile:

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OfflineMushyMom
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Re: Diagnosing contamination issues/reviewing technique [Re: MushyMom]
    #28241498 - 03/22/23 02:17 PM (1 year, 9 months ago)

Prepped Millet yesterday. Added to boiling water.  Covered and took off heat for an hour. Loaded jars about halfway full. Pc - vent and then 15 psi for 2 hours.  Note to self - millet expands when it cooks.


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Invisibledowodenum
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Re: Diagnosing contamination issues/reviewing technique [Re: MushyMom]
    #28241508 - 03/22/23 02:22 PM (1 year, 9 months ago)

Yeah, it definitely does. Think we've all made that mistake!

I suggest weighing your jars (after taring out an empty one), getting them all the same ish, then writing on the jar what the load weight was. You only really need to mark one jar, if they all act the same. Having a number to go by will help you titrate to land on 3/4 full, over time.

I've also noticed that top layer jars (sideways) don't plump up quite as much as bottom layer. Something about the surface area maybe.

If you switch suppliers, you might have to do it again, due to different initial ("dry") hydration amounts.


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OfflineMushyMom
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Re: Diagnosing contamination issues/reviewing technique [Re: MushyMom]
    #28249714 - 03/27/23 05:53 PM (1 year, 9 months ago)

3-27 11 days after shaking - This is the WBS that was inoculated on 3-3 and shaken on 3-16.  These are 100% bacterial.  I might try top fruiting.  Not sure yet.
Quote:

MushyMom said:
Prepped grain jars - using WBS - added to boiling water, took off heat and let soak for 90 minutes.  Drained.  Loaded jars.  PC’d in jars with unmodified plastic lids (vented for 15 minutes then PC for 90).

3-3 Used LC from Shroomery sponsor.  Applied to agar at the same time as the jars to test. Plates were clean. 

3-16 - shook jars

These pictures were taken last night





I’m not nuts about the way the grain looks.  I’m in wait and see mode on these.

Concerns are
1. Old WBS
2. Unmodified lids - some are warping.













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OfflineMushyMom
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Re: Diagnosing contamination issues/reviewing technique [Re: MushyMom]
    #28249718 - 03/27/23 05:55 PM (1 year, 9 months ago)

These are the other 4 jars - I don’t think they look as bad as the ones in the last post but I’m still not nuts about them.  Might shoebox them


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Offlineside-eye
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Registered: 09/01/22
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Re: Diagnosing contamination issues/reviewing technique [Re: MushyMom]
    #28249727 - 03/27/23 06:02 PM (1 year, 9 months ago)

Quote:

MushyMom said:
These are the other 4 jars - I don’t think they look as bad as the ones in the last post but I’m still not nuts about them.  Might shoebox them






Also consider [Fahtbags] as an option. I've heard it's a bit easier to maintain moisture in them compared to shoeboxes.


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OfflineMushyMom
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Re: Diagnosing contamination issues/reviewing technique [Re: side-eye]
    #28249731 - 03/27/23 06:04 PM (1 year, 9 months ago)

I have unicorn bags for fruiting. I tried using them in the past but everything went.straight to contamination and didn’t pass go or collect $200…. I’m considering it - it seems like it would be easier to contain contam in the bag if it does go green again?

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OfflineMushyMom
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Re: Diagnosing contamination issues/reviewing technique [Re: MushyMom]
    #28254945 - 03/30/23 03:46 PM (1 year, 9 months ago)

Just for shits and giggles I decided to open plates at my desk area (where I’ve been using my SAB lately) and in my closet (where I used to use the SAB and where I still store everything like my grain jars and Petri dishes. 

I opened these 3 days ago - gave them a quick wave and then closed them back up. So far the closet dish is taking off and the desk dish hasn’t sprouted anything - yet.

I usually keep my bins in my closet but I’m wondering if I should keep them somewhere else now. I also suspect having a couple bins go green in the closet in the past probably didn’t help much


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Invisibledowodenum
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Re: Diagnosing contamination issues/reviewing technique [Re: MushyMom]
    #28254960 - 03/30/23 03:54 PM (1 year, 9 months ago)

Yeah closets (with clothes in) are pretty bad, I'd sooner use a clean kitchen any day.


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OfflineMushyMom
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Re: Diagnosing contamination issues/reviewing technique [Re: dowodenum] * 1
    #28254963 - 03/30/23 03:58 PM (1 year, 9 months ago)

Quote:

dowodenum said:
Yeah closets (with clothes in) are pretty bad, I'd sooner use a clean kitchen any day.



Yeah it sucks. I’m a grown ass adult and I own my own home and yet I’m confined to the closet lol.

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OfflineMushyMom
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Re: Diagnosing contamination issues/reviewing technique [Re: MushyMom]
    #28259355 - 04/02/23 03:47 PM (1 year, 9 months ago)

Mixing substrate and moving to fruiting.

These are from the first batch - wbs with lc purchased from a shroomery supplier. Jars with unmodified lids.

4 seemed really sus along the way. The other 4 were slightly sus. I marked the jars and will be comparing results along the way

I will be doing 2 shoeboxes and 2 unicorn bags of each. I am thinking of using a 1:1 ratio with the more sus jars and a 2:1 ratio on the better jars.  Prepping everything now

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OfflineMushyMom
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Re: Diagnosing contamination issues/reviewing technique [Re: MushyMom]
    #28259677 - 04/02/23 07:18 PM (1 year, 9 months ago)

These were the really bad jars


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