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InvisibleLenz
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Insane Contamination Rates on Grain Spawn. Looking For Help.
    #27121551 - 01/02/21 05:41 AM (3 years, 26 days ago)

Hi all, I'm a bit desperate for advice right now.

So after a few nice months of experiencing nearly 0 contamination with my grain spawn, I’m now finding over 50% of my spawn contaminating either before inoculation (having sat anywhere from a few days to a week post-sterilization) or shortly after inoculation via LI. Nothing about my process or my grain has changed, same old blended LI to Fooman prepped Pennington's WBS. 

Pics of the contaminant are below.

Pertinent details:

- I use Pennington’s classic WBS and PC at 16-18PSI for 2 hours.

- For inoculation I always go 250-300mL blended LI per 10 jars.

- Plastic lids have SFD’s siliconed on, lids left just barely loose during PC.

- SAB is sprayed down and air allowed to settle between every batch of jars inoculated.

- The contamination is mostly a green mold or black pin mold that starts on a few grains spread throughout the jar.

- Contamination is difficult to isolate to a single vector, being present through multiple batches of jars, some with brand new SFD's and some with older SFD's, across different cultures and plates, and different WBS prep methods (Fooman and no prep).


What I'm really struggling with is pinning down the source of the contamination. It seems unlikely that my inoculating procedure in the SAB has gotten worse since I started, so I don't think that's it. Furthermore, the contamination is present on jars that have brand new SFD's on the lids and older SFD's. I don't suspect my cultures either, as they all look nearly pristine and the jars that have contaminated have been all different cultures and varieties. 

I’m venting properly and anything longer than 2 hrs seems overkill but I'll try upping it 2.5 hours.

I was leaning towards the idea that my filters are failing somehow but now with even more jars having been contaminated, it really seems unlikely that this many SFD's are suddenly failing out of nowhere.

The only thing that makes sense is that somehow that 2 hours isn't cutting it anymore and that I need to up the PC time or that I'm not properly sterilizing somehow.


Apologies for the lengthy post but like I said, I'm at a loss for why this is suddenly happening. Has anyone dealt with anything like this before as well? Could it be this batch of WBS needing more than 2hrs?

How the hell do I stop this?



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InvisibleModularMind
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Re: Insane Contamination Rates on Grain Spawn. Looking For Help. [Re: Lenz]
    #27121593 - 01/02/21 06:32 AM (3 years, 26 days ago)

How do you prep your LI?


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InvisiblebodhisattaMDiscordReddit
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Re: Insane Contamination Rates on Grain Spawn. Looking For Help. [Re: ModularMind]
    #27121594 - 01/02/21 06:35 AM (3 years, 26 days ago)

Mold contamination dies before the pressure cooker gets to pressure. Mold contamination is indicative that the cultivator fucked up in technique.

Skip LI until you get your skills honed.

Increasing your PC time won't fix shit. Bacterial endospores are the only reason we sterilize so long. Molds die very easily


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InvisiblePsicomb
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Re: Insane Contamination Rates on Grain Spawn. Looking For Help. [Re: bodhisatta]
    #27121626 - 01/02/21 07:08 AM (3 years, 26 days ago)

Why would their jars molding before inoculating as well tho?

Wanna take pics of your filters too, OP?

I do agree LI is not ideal.

Are you venting your PC for a handful of minutes before putting on the rocker?


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When we constantly pull things apart trying to see how it works, we may end up with only an understanding of how to destroy something
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InvisibleLenz
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Re: Insane Contamination Rates on Grain Spawn. Looking For Help. [Re: Psicomb]
    #27121902 - 01/02/21 11:42 AM (3 years, 26 days ago)

Thanks for the replies all.

@ModularMind, Oster blender attachment siliconed onto the underside of the jar band with a rubber gasket ring siliconed under that. PC’d for 25 minutes at 16-18 PSI. I let it cool then carefully transfer 2 halves of a culture in 2 motions before blending.

@Bod, That’s a good point, thank you. It’s possible that it’s my technique but the fact that I’ve gone a few hundred jar runs without anything resembling this level of contamination makes me a little bit skeptical. I feel like I would’ve had to have had some seriously fucked SAB sessions to get this much mold, not to mention that a few jars molded before I even opened them. I did jump in head first to LI however so I will take a look at what could be going wrong there.

@Psicomvb, agreed on the point about molding before inoculation. At this point I do think it’s either my filters or something going on with my LI or both. I’ll likely remake most of my filters and re silicone the LI blender apparatus. I’ll go A2G if I really have to but LI is a bit hard to give up in my current workflow.

And pics of filters:



Definitely should’ve sanded the lid more and spread the silicone around a bit more to prevent peeling but they’ve been just fine before.


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Offlinejcm4620
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Re: Insane Contamination Rates on Grain Spawn. Looking For Help. [Re: Lenz]
    #27121920 - 01/02/21 11:56 AM (3 years, 26 days ago)

are u even using agar??


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InvisibleLenz
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Re: Insane Contamination Rates on Grain Spawn. Looking For Help. [Re: jcm4620]
    #27121937 - 01/02/21 12:04 PM (3 years, 26 days ago)

Quote:

jcm4620 said:
are u even using agar??




?? How would I make LI without agar??


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OfflineTheBoJim
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Re: Insane Contamination Rates on Grain Spawn. Looking For Help. [Re: Lenz]
    #27121959 - 01/02/21 12:17 PM (3 years, 26 days ago)

maybe its your grain. someone on here told me that bad batches of grain are a thing. did you start using a new bag of seed when the problems started?


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Offlinejcm4620
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Re: Insane Contamination Rates on Grain Spawn. Looking For Help. [Re: TheBoJim] * 1
    #27121969 - 01/02/21 12:18 PM (3 years, 26 days ago)

lol ive seen ppl make a Ll by shooting spores into shit so i had to ask.


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OfflineTheBoJim
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Re: Insane Contamination Rates on Grain Spawn. Looking For Help. [Re: jcm4620]
    #27121971 - 01/02/21 12:20 PM (3 years, 26 days ago)

Quote:

jcm4620 said:
lol ive seen ppl make a Ll by shooting spores into shit so i had to ask.




guilty :grin:


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InvisibleLenz
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Re: Insane Contamination Rates on Grain Spawn. Looking For Help. [Re: TheBoJim]
    #27122055 - 01/02/21 12:56 PM (3 years, 26 days ago)

@BoJim, I was thinking it could be my grain as well, the contamination somewhat roughly aligns with some new bags of WBS I got that were quite a bit dustier and dirtier than usual, but like Bod said, I’m not sure how molds would survive a 2 hr PC cycle even if the grain was dirty as hell.

And @jcm, lol yeah not here fortunately. Probably would’ve experienced something like this a long while ago if I’d been shooting spores this whole time.


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OfflineTheBoJim
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Re: Insane Contamination Rates on Grain Spawn. Looking For Help. [Re: Lenz]
    #27122082 - 01/02/21 01:15 PM (3 years, 26 days ago)

endospores, not mold. Doesn’t hurt to get new grain.


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InvisibleLenz
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Re: Insane Contamination Rates on Grain Spawn. Looking For Help. [Re: TheBoJim]
    #27122090 - 01/02/21 01:20 PM (3 years, 26 days ago)

But even if endospores survived, there shouldn’t be any mold surviving to germinate, bacterial presence or not, no? And I’d get new grain but I’m stuck with about 80lbs of this batch right now, would rather try and find a way to use it.


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OfflineTheBoJim
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Re: Insane Contamination Rates on Grain Spawn. Looking For Help. [Re: Lenz]
    #27122106 - 01/02/21 01:29 PM (3 years, 26 days ago)

If you were trying to eliminate the source of you problems and you think grain might be an issue then getting a different (small/cheap) batch could be a good idea.


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InvisiblePsicomb
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Re: Insane Contamination Rates on Grain Spawn. Looking For Help. [Re: TheBoJim] * 1
    #27122128 - 01/02/21 01:42 PM (3 years, 26 days ago)

It isnt the grain man.  It isn't being properly sterilized, or contams are getting into the jar post-PC.  It is that simple.

Are you letting it vent before putting the rocker on?

It will read a higher psi than it is if you do not vent ahead of time. This will lead to contams in jars

Something is very wrong if you get contams before inoculating. 


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

When we constantly pull things apart trying to see how it works, we may end up with only an understanding of how to destroy something
- nick sand


Edited by Psicomb (01/02/21 01:43 PM)


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InvisibleLenz
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Re: Insane Contamination Rates on Grain Spawn. Looking For Help. [Re: Psicomb]
    #27122146 - 01/02/21 01:53 PM (3 years, 26 days ago)

Quote:

Psicomvb said:
It isnt the grain man.  It isn't being properly sterilized, or contams are getting into the jar post-PC.  It is that simple.

Are you letting it vent before putting the rocker on?

It will read a higher psi than it is if you do not vent ahead of time. This will lead to contams in jars

Something is very wrong if you get contams before inoculating. 





I’m with you on that, don’t see how molds could be surviving 2 hrs in the PC.

And yeah, forgot to answer that, I always vent for at least 10 minutes.


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InvisiblePsicomb
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Re: Insane Contamination Rates on Grain Spawn. Looking For Help. [Re: Lenz]
    #27122199 - 01/02/21 02:31 PM (3 years, 26 days ago)

Well, if it is rocking and the PSI is accurate it must be your filters.  What percentage would you say contams before you inoculate?  Does it seem to happen more with the inoculated jars than the ones that arent inoculated?

Does the contam start on the top of the grains and work it's way down? That's a way to determine if the filter is the source. Use the location of the contam to try and speculate how or where it originates

Sorry to hear about this, that sucks man.  Unexpected contams really can be frustrating.


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When we constantly pull things apart trying to see how it works, we may end up with only an understanding of how to destroy something
- nick sand


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Offlinebw86
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Re: Insane Contamination Rates on Grain Spawn. Looking For Help. [Re: Psicomb]
    #27122253 - 01/02/21 03:04 PM (3 years, 26 days ago)

like others said its 100% not the grain.


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OfflineMycoBrainz
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Re: Insane Contamination Rates on Grain Spawn. Looking For Help. [Re: bw86]
    #27122293 - 01/02/21 03:24 PM (3 years, 26 days ago)

I have a 941 so I kinda have to go at least 2.5 hrs to avoid contamination. When I do bags I roll min 3 hrs. Then I wait till it cools to open. I just ran 4 qt jars and 9 spawn bags. It took 3.5 hrs. No contamination yet. Once in a while I get a bad syringe. To test use a little on multiple batches. If they all contaminate when others grow fine its the mss. Rollin 19 qt at a time is nice.


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InvisibleJosex
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Re: Insane Contamination Rates on Grain Spawn. Looking For Help. [Re: MycoBrainz]
    #27122301 - 01/02/21 03:30 PM (3 years, 26 days ago)

It does sound like technique or filter failure, but I'm betting the ranch on the latter.


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OfflineGypsyBastard
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Re: Insane Contamination Rates on Grain Spawn. Looking For Help. [Re: Josex]
    #27122313 - 01/02/21 03:37 PM (3 years, 26 days ago)

I feel like, as others have said, it can only be 1 of 2 things. If jars are contaminating after PC but before being opened, then either
A. The sterilization process was not adequate
B. The filters are not adequate.

I would PC a jar with no filter, no poly, no MP. Just a jar of grain and a lid. Then wait. If this jar contam, it is your sterilization. If not, it's the filters. Take sometime, narrow it down to this one variable and you will have your answer and can make corrections as needed.
Like others have said, it doesn't matter what grain you are using, if things are growing this its not sterile. You just need to find out if it ever was.


Edited by GypsyBastard (01/02/21 03:38 PM)


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InvisibleLenz
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Re: Insane Contamination Rates on Grain Spawn. Looking For Help. [Re: GypsyBastard]
    #27132127 - 01/07/21 02:26 AM (3 years, 21 days ago)

Apologies for the late response, I just wanted to say thanks to everyone who chimed in, it's much appreciated, thank you all.

I've been thinking about it some more and I believe it's most likely a combination of filter failure and bad technique.


I say this because:

- I've had a couple jars contam prior to inoculation, but the majority occurs or is noticeable after inoculation. It also seems unlikely that so many of my filters would all happen to fail within a few weeks of each other.

- While I said in the OP that I haven't changed my process at all, I realized that I had started stirring my LI more frequently while pouring in order to distribute the myc more evenly.
I'm not exactly sure this could cause this much contamination, but all that air movement with the LI jar wide open wasn't too bright of me.

- I've been careless about how I shake my jars right after inoculation as well, with 25-30mL of LI I should be much more careful about not getting the SFD too wet. (Not sure if this is a real issue, from reading it seems that synthetic non cellulose filter disc's shouldn't be too susceptible to a little moisture as long as they dry out quickly).



For the time being, for peace of mind, I've re-made a bunch of lids with new SFD's, re-siliconed my LI blender attachment, PC'd 4 test jars that I'll leave sitting for a few weeks, and am planning to tighten up my SAB movement.

I think it was my mistake to have the level of confidence in my SAB procedure from having had a couple months of success. I think the fact that I had some jars contaminate prior to inoculation also skewed my perception of what was going on.
While I am still relatively confident about my SAB work, I started doing things without really thinking just because I'd been doing ok previously.
Inoculating 30-40 jars a week, I definitely got a bit lax in some ways and that certainly played a part in my recent issues.
Vigilance must be maintained lmao.

Anyways, I'll be inoculating a bunch more jars soon so I'll see how those do and hopefully won't be running into too many more issues.


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