Let's Talk Grain Prep: The Good, The Bad and The Hardy!
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tedoro
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Josex
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tedoro
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tedoro
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tedoro
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tedoro
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tedoro
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tedoro
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Josex
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Josex
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Josex
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Josex
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Josex
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Dendrocopos
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tedoro
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Dendrocopos
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Josex
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Josex
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fahtster
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Josex
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Dendrocopos
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Josex
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QM33
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Dendrocopos
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Josex
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Josex
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05/12/22 05:06 PM
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Hindsight
Mad Scientist
Reged: 01/24/21
Posts: 2706
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Re: Let's Talk Grain Prep: The Good, The Shitty and The Hardy!
05/15/22 07:58 AM
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Great test in the OP. I followed it last year but forgot to chime in so I could see all the updates. Now I have 26 pages to read back through.
But, reading the first page updates and scanning through the last pages, I have a hunch I might know what is going on here. Sorry if this has already been mentioned in an earlier post that I haven't yet read through.
There are substances produced by various bacteria that, even after the bacteria are dead, remain active. These substances are part of their evolutionary survival mechanisms, and serve to kill other organisms that compete with the bacteria. One that immediately comes to mind is spinosyn which is sold under the brand name Spinosad as a pesticide. Another which isn't a single substance but is a more general description for inhibiting substances produced by bacteria is called Bacteriocins (which are peptides), "bacteriocin-like inhibitory substances" or "BLIS" for short.
Josex'worst performing grain batch was fermented. People have fermented foods for thousands of years as a means of food preservation. This article specifically calls out lactic acid bacteria (the bacteria responsible for fermentation) as being especially skilled at producing peptides that fight food spoilage. And while this article focuses on the bacteria inhibiting properties of these peptides, other studies have been done on these same peptides that prove their inhibiting effects on spore viability and mycelium growth as well! https://pubmed.ncbi.nlm.nih.gov/30448894/
Here is another: https://www.frontiersin.org/articles/10.3389/fmicb.2016.00461/full
Here is one study proving the inhibitory affects Lactobacillus peptides have on molds and spores: https://microbiologyjournal.org/control-of-food-spoilage-molds-using-lactobacillus-bacteriocins/
I believe the reason for poor grain performance in Josex' first batch is due to the peptides produced by lactobacillus during the grain soaking process. I also think that how grain is stored prior to use (by the farmer, or anywhere in the supply chain) can have an impact on the type and amount of bacteria that grows on the grains, and as such, the peptides left behind. And while I haven't searched for any articles on the possibility of various fungus/molds leaving their own peptides behind that could negatively impact the growth of mycelium, I would guess that this too is a strong possibility.
So it stands to reason that once these substances have been produced, they are much more difficult if not impossible to remove than live bacteria, bacterial endospores, or mold spores. I did see the posts with some people saying that it is ok to use grain water agar made from the prep of grains that provided poor performance, but I think that might need more exploration/testing to be conclusive (for example, I would love to see two batches of grain water agar tested side by side - one batch made from water from fermented grain prep and the other made from water from standard grain prep). It is also possible that part of this is due to the better heat conductivity of water vs hydrated grains. But the main question is: How do you get rid of mycelium inhibiting peptides once they are produced? I googled "Bacteriocin heat stability" and took a quick look at some results. A lot more time should be spent but in one example, I found this, specifically referring to bacteriocins produced by lactic acid bacteria:
"The bacteriocin did not lose anti-listerial activity when treated at 100°C for 30 min or at 121°C for 15 min. The bacteriocin lost its antimicrobial activity after treating with trypsin, protinase-K, protease and peptidase." https://pubmed.ncbi.nlm.nih.gov/23100837/
That's the only study I have looked at so far, it didn't say if they went higher than 121C, and it didn't say if they were using pressure as well (I assume they weren't). The main point is that these peptides appear to be at least somewhat heat stable so it's possible that a PC cycle isn't enough to break them down. But note the treatments that were able to break them down. Trypsin is a digestive enzyme that you can easily buy. Proteinase-K (typo in the abstract above) was discovered as something produced by fungus and can be purchased from lab supply stores: https://en.wikipedia.org/wiki/Proteinase_K.
Since we don't know for sure if the inhibiting substances impacting our grain jars is only produced by bacteria, or can be produced by bacteria and fungus/molds, some tests would need to be done. I think Josex' fermentation grain prep is pretty conclusive that bacteria-produced peptides are inhibiting mycelium growth. It would be interesting to do another fermentation grain prep test, then treat the soaked grain with trypsin or proteinase-k, then PC the grain and see if anything changes. A lot of variables around that though.... how much to use, how long to soak, would risidual trypsin have a negative impact mycelium etc. With more research, I think a lot of those answers could be found ahead of time.
-------------------- * How eliminate condensation from no-pour agar plates * Build a steam pasteurizer * Learn about trich
     
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Re: Let's Talk Grain Prep: The Good, The Shitty and The Hardy!
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tedoro
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