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OfflineAgarStudent
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Is this bacterial?
    #23880839 - 11/30/16 07:29 AM (7 years, 1 month ago)

Some of my small spawn bags of galindoi have done this:



Is this definitely bacterial?  Too much water or not enough pressure cooking to kill them?

Cheers


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InvisibleTeemo 6T3
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Re: Is this bacterial? [Re: AgarStudent]
    #23881378 - 11/30/16 11:54 AM (7 years, 1 month ago)

Yup, its bacteria.

Did you start the spores on agar?

How long did you pressure cook it?

Tell us more about your procedure so we can help you out more and see how you can prevent this in your upcoming grows.


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OfflineAgarStudent
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Re: Is this bacterial? [Re: Teemo 6T3]
    #23882129 - 11/30/16 03:44 PM (7 years, 1 month ago)

Spore to agar to strain number reduction/ isolation. Then to jars of grain to spawn bags.  Small spawn bags were PC for about 1.5 hours. Cheers


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InvisibleTeemo 6T3
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Re: Is this bacterial? [Re: AgarStudent]
    #23883932 - 12/01/16 08:59 AM (7 years, 1 month ago)

So you got it running on agar, no contaminants observed right?

If so this would be due to not sufficient time in the PC, bags need more time in the PC because the proper sterilisation heat needs to get to the middle of the bag too, not like in grain jars.

Quote:

Then to jars of grain to spawn bags.




I don't get what you mean by this.

Do you mean you transferred the agar to grain jars, and then did a G2G (grain 2 grain) to spawn bags?

If so then it could of also contaminated due to improper sterile procedure. Did you do it in a SAB?


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OfflineAgarStudent
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Re: Is this bacterial? [Re: Teemo 6T3]
    #23884627 - 12/01/16 02:10 PM (7 years, 1 month ago)

Sure, that was G2G from colonised jars, but over the laminar flow hood to ensure sterile air.
Not all of the bags went bad - mostly only a few and consistently from the centre.  This makes me think it was insufficient PC time? Plus my PC was on the way out and not getting good and consistent pressure.


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Offlinekatbusa
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Re: Is this bacterial? [Re: AgarStudent]
    #23889927 - 12/03/16 08:35 AM (7 years, 1 month ago)

Quote:

AgarStudent said:
Spore to agar to strain number reduction/ isolation. Then to jars of grain to spawn bags.  Small spawn bags were PC for about 1.5 hours. Cheers




I'm sure you know this, so please don't take offense.

Did you soak your grain over night to allow the endospore bacteria to germinate?

If you are using .2 micron bags try not to seal them before you PC them. You can damage the filter but I have head of people getting away sealing the .5 micron bags.

Your grain looks wet but over all though I think you are right about your PC going on the fritz.


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OfflineAgarStudent
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Re: Is this bacterial? [Re: katbusa]
    #23889999 - 12/03/16 09:19 AM (7 years, 1 month ago)

Definitly an overnight soak and never sealing the bags until after PC.  Otherwise I can't get grain in there :wink:. Next time I PC for two hours minimum, 15psi


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Offlinekatbusa
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Re: Is this bacterial? [Re: AgarStudent]
    #23890286 - 12/03/16 11:17 AM (7 years, 1 month ago)

Quote:

AgarStudent said:
Definitly an overnight soak and never sealing the bags until after PC.  Otherwise I can't get grain in there :wink:. Next time I PC for two hours minimum, 15psi




Yea I I had a feeling you knew what you were doing. Upping the PC time sounds like a good plan!

Good Luck!


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Invisiblevatman
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Re: Is this bacterial? [Re: katbusa]
    #23891753 - 12/03/16 07:26 PM (7 years, 1 month ago)

Did you have the grain sitting on the filter patch?

The filter patch will whick contamination.

I run my bags for 2-2.5 hours.

Also looks like high water content.


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OfflineAgarStudent
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Re: Is this bacterial? [Re: vatman]
    #23895739 - 12/05/16 01:21 AM (7 years, 1 month ago)

Ah I sit the bags flat in order to try and maximise the surface area of the grain that is in contact with the plastic.  This is because stone formation typically occurs at non-nutritious boundaries.

Grains are pretty much in contact with the filter patch, but it is 0.1 micron, so stuff cannot get in or out besides air.  How is it that contamination can wick through?
Should they be upright with filter clearance and not laying flat?

From now on 2.5 hours minimum on the pressure cooker, having just replaced my old Fagor Duo with a Swiss made Kuhn Rikon (awesome bit of stainless gear that so far works a treat).
Sometimes with the cheaper cookers the pressure is unreliable, especially when the gaskets are done.

Thanks again for your help.


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Edited by AgarStudent (12/05/16 02:20 AM)


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Invisibleegobrain


Registered: 11/17/16
Posts: 180
Loc: Canada Flag
Re: Is this bacterial? [Re: AgarStudent]
    #23897651 - 12/05/16 05:54 PM (7 years, 1 month ago)

Maybe your moisture content was thrown off during PC'ing. The patch isn't going to wick contams. It seems to me that your grains were just too wet.


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OfflineAgarStudent
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Re: Is this bacterial? [Re: egobrain]
    #23898561 - 12/05/16 10:44 PM (7 years, 1 month ago)

I think so?  How long do you boil grains for compared to Mason jars, where I would boil for about 10 min.


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