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InvisibleWilly Wonka
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Causes of a bacteria contamination?
    #22316008 - 09/30/15 07:50 PM (8 years, 3 months ago)

Hello gang.  I've been fighting bacteria contamination quite a bit.  I have experienced bacteria contamination with various strain from various syringes, from various places and in multiple grain jars.  For the most part all of my jars look really good and clean up until almost the end and then they start to show signs of bacterial contamination.  I always pretreat my grains with a 24 hour soak.  After the soak I bring the soak water to a full boil then drain.  I always lay the grains out on a work table and allow the grains to fully dry (tissue test) before I load them into their jars.  Then the jars all get PC'ed at 15 PSI for 90 minutes. Sounds like standard protocol right?

Okay now that you know the prep let me tell you a bit about the colonization conditions.  The jars are allowed to colonize in my garage.  I have been monitoring the temperatures and on the high end of things it gets into the high 90s during the day. 

Now my contamination questions.  Is bacteria primarily a grain issue or can you get spores that are contaminated from the start?  I know you can get spores that are contaminated with just about anything, but I keep running into bacterial contamination from multiple syringes, from multiple places.  This leads me to believe that my contamination vector is not the spores.  I am also pretty sure that my grain preparation is adequate to kill any bacteria endospores that might be present in the grains prior to inoculation.  Can bacteria contamination be caused by grain problems?  Can the bacteria be from the temperatures being to high and/or the grains being to wet?


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Invisiblemicro
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Re: Causes of a bacteria contamination? [Re: Willy Wonka]
    #22316106 - 09/30/15 08:11 PM (8 years, 3 months ago)

high 90's is way too high

that will favor bacterial growth, as would too much water

i agree it is probably environmental but i'm not sure the vector

possibly inoculation ?  just a guess

everything should have been sterilized at those params


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InvisibleWilly Wonka
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Re: Causes of a bacteria contamination? [Re: micro]
    #22318481 - 10/01/15 10:43 AM (8 years, 3 months ago)

Yes I agree.  Those temperatures are way too hot.  I am trying to make a cooling system for my garage.  I know the bacteria isn't introduced during inoculation.  I flame sterilize my needle for every jar, and I work in front of a flow hood. I don't think there is too much water in the jars before I inoculate them  because I do the toilet paper test on my grains.  Although I do see an extreme amount of condensation in the jars at peak temperatures.  This leads me to believe perhaps the high temperatures might be pulling all the moisture from the grains, and then the moisture condenses in the jars causing really wet conditions outside the grains.  So my thought is that perhaps this combined with the high temperatures is the reason for the bacteria contamination I have been getting?


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Invisiblemicro
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Re: Causes of a bacteria contamination? [Re: Willy Wonka]
    #22318577 - 10/01/15 10:59 AM (8 years, 3 months ago)

it could be a factor...

it's a combination of lots of things

if you get everything optimal as you can you will have the most success

also, that doesn't really exlain where the spores came from, that's still an unknown

i'd look at where it is in the jar; might give you a clue


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OfflineKizzle
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Registered: 08/30/11
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Re: Causes of a bacteria contamination? [Re: micro]
    #22319627 - 10/01/15 03:14 PM (8 years, 3 months ago)

Quote:

Is bacteria primarily a grain issue or can you get spores that are contaminated from the start?



If you use spores that were recently dropped there's a high chance of bacteria. Most bacteria will dry out and die a few days after making a print so you should wait before using one to make a syringe. Of course not all bacteria die from desiccation and they can also be introducing while making the syringe.

Bacteria can survive insufficient sterilization times. Excess water can create an environment that allows anaerobic bacteria, which includes some of the most heat resistant species and mostly likely to survive sterilization to grow. They will often be accompanied by a fermentation/alcohol type odor. High temperatures can also allow certain heat resistant bacteria to grow that would  have otherwise reproduced very slowly when present.


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