• Dead LAB is a source of food for some higher fungals.
  
  
  
• LAB has a evolutionary synergistic relationship to many higher fungals and lower fungals (microbes) as LAB is common in many places and in media where higher fungals grow in Nature.  Anaerobic organotrophic bacteria is thought to be part of the "primordial soup" which has given rise to the evolutionary process to todays date...
  
  
  
• LAB excretes enzymes which ferment grain, bulk subs, etc (organic matter).  Releasing 'bio-nutrients' while the enzymes ferment the media and lower the pH.  The fermentation also softens grain/WBS/etc and may assist mycelium in colonizing grain/WBS/etc.
  
  
  
• LAB will colonize media and help to 'block' the colonization of the media by harmful microbes.  This is accomplished mainly by LAB's production of lactic acid which "...is a strong sterilizing compound, and [LAB] suppresses harmful microorganisms and enhances decomposition of organic matter. Moreover, lactic acid bacteria promote the decomposition of material such as lignin and cellulose and ferments these materials...".
  
  
  
• LAB offers zymogenic balance in media such as bulk sub due to it's fermenting ability, etc:
  http://www.agriton.nl/higa.html#21
  

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  Zymogenic Soils:
  These soils are dominated by a microflora that can perform useful kinds of fermentations, i.e., the breakdown of complex organic molecules into simple organic substances and inorganic materials. The organisms can be either obligate or facultative anaerobes. Such fermentation-producing microorganisms often comprise the microflora of various organic materials, i.e., crop residues, animal manures, green manures and municipal wastes including composts. After these amendments are applied to the soil, their number: and fermentative activities can increase dramatically and overwhelm the indigenous soil microflora for an indefinite period. While these microorganisms remain predominant, the soil can be classified as a zymogenic soil which is generally characterized by a) pleasant, fermentative odors especially after tillage, b) favorable soil physical properties (e.g., Increased aggregate stability, permeability, aeration and decreased resistance to tillage c) large amounts of inorganic nutrients, amino acids, carbohydrates, vitamins and other bioactive substances which can directly or indirectly enhance the growth, yield and quality of crops, d) low occupancy of Fusarium fungi which is usually less than 5 percent, and e) low production of greenhouse gases (e.g., methane, ammonia, and carbon dioxide) from croplands, even where flooded rice is grown.
  
  

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One aspect I really like is the possibility of using LAB (or a BIM mix) within casing to prevent harmful microbes, trichoderma, etc.  This could mean the pH may not have to be increased in casing which should be beneficial for mycelium that often prefers a slightly acidic pH of around 4.5-6. 

Up next: Step-by-Step Guide

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quickpick said:
Hey Cakes,

Yea you could buy a PC but where's the fun in that?
Later

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thank you for collecting information and providing step by step instructions for culturing LAB! As soon as I find the time, I will try myself. I'm quite curious about the fermentation of horse manure.

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You welcome.  Yea me too, LAB just loves manure

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Regarding fermentation of straw I found some interesting PDF.
'Effects of Water-Soluble Carbohydrate Content on Silage Fermentation of Wheat Straw'
http://www.jstage.jst.go.jp/article/jbb/101/3/101_232/_article
They tested the effect of various amounts of added sugars on PH, lactic acid and side products. Seems like 7 to 10% sugar (compared to dry mass) works best.

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Nice study, thanks!

Yes that's sounds right, but they are using glucose so I assume measuring by weight.  That's good to know but hard to put into practice when one would first need to know the DW of the matter to be fermented.  Another thing which would make this difficult to put into practice is one would first need to know the inherent WSC of the matter to be fermented.  Then glucose is added, taking into calculations the inherent WSC content to make the 'new' (for lack of a better term) WSC content equal  >7-10% of DW.  So if I understand the methods correctly it would be hard to impossible for average people to manipulate the levels of WSC as per the study.

Another point worth mentioning is what they are doing is more similar to what you did with your grain thread.  They are relying on "endogenous" microbes (LAB) from the wheat straw (and air too?) to naturally ferment the matter, they did not apply an already highly active LAB 'pure culture'.  When they talk about adding glucose it's to feed the LAB because the inherent WSC of the wheat was not sufficient to feed the LAB so they could multiply and ferment.  This is why I originally suggested to you that you may want to add black strap molasses to your jars while fermenting your grain.

What we are doing with pure cultures (LAB, BIM mixes, AEM, etc) is adding microbes (LAB in this case) in massive quantities which are already being feed and are very active, releasing enzymes, lactic acid, etc.  We don't have to rely upon endogenous LAB so the process of fermentation is generally faster and more efficient.  As is the LAB's ability to 'block' harmful microbes by colonizing the media really fast, before other microbes get the chance to take hold, and by virtue of lactic acid.

For use with LAB and other BIM (AEM, FPE, etc) a suggested 'benchmark' (if you will) is a brix level of 9-11.  And a 1:1:20 ratio with black strap molasses (80-90 brix) is approximately 9-11 brix.  A higher brix level will slow the pH drop and is used in special applications a bit out of scope of this thread.  When a brix level of 11 is used there is a good bit of carbohydrates left over after the pH drops below 3.5. (when the BIM is ready to use).  The left over carbohydrates serve as food for the microbes which assists them while fermenting matter like grain, etc.  So in other words when we ferment a BIM or AEM with black strap molasses we doing the same thing (albeit in a different manner) as the study above:  Increasing the quantity and effectiveness of LAB and their fermentations.

HTH

Hey CnO,

Nice nick

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Not to mention that experiments like these are hardly ever very useful at the beginning.  What we need is a proof a principle (which you are doing here) and then the kinks get worked out and then BAM.  Suddenly you got people growing 8 foot cubes.

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There is a lot to be learned from failure, more then from success in most cases IMVHO .  That said, I do have high hopes and high expectations, but I'm also expecting failure. 

Man, how amazing would that be?  8 foot tall cubes, lol.  Like Alice in Wonderland!

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Good job.  If you need any lab stuff let me know.  If I have it I'll ship it to aid in the process.

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Thanks. 

Well now you mention it, I do have a shopping list.  If you have any items you could send that would be great!

Most items are for a control BIL (Basic Isolation Liquid) I'm not using "Basic Isolation Media", aka "BIM" because it too confusing with the other term of BIM.  While testing isolation of PnSB from Natural water with my DIY method I want to use a proven isolation method as a control.  Though I've already provided proof of concept and collected water from Nature, isolated, and then cultured PnSB with hydrolyzed fish and distilled water. (that thread will be part 3 of my "BIM" series).



Shopping list:

• (NH₄)₂SO₄
  
  
• K2HPO4
  
  
• MgSO₄
  
  
• NaCl
  
  
• Yeast extract
  
  
• Glycerol

Thanks

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sure, I realized they were doing the same as you by adding molasses, just thought it would be interesting because of their measurements of side products, especially alcohol and carboxylic acids. That study on straw mentions an initial sugar content of 1.4%. If we assume dry straw to have a water content of 5 - 10%, it is not too hard to estimate roughly how much sugar to add.

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As grain should have a sugar content of around 10%, I favor fermenting without adding sugars.

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I only wonder about the initial sugar content in horse manure...

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Yea, and it's pretty variable depending upon what they ate and the WSC content of the food they ate.


My 2 cents..

Thanks buddy!

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Nice thread there bro,

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C? Is that you?
(with you, springs and I over here it's like a little CW)

If you are who I think you are then forgive me when I post a little bit of 'basic' info within my response to you.  The info is for the greater audience reading, not for you per say.

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I'm going to add my 2 cents here:

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We know that an AEM culture  of 1:1:20 uses about 4.5% molasses by volume and an AEM culture  of 1:1:10 uses about 8.5% molasses by volume so I think it is safe to say if we can get the initial molasses volume in that range then we will be good to go.

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Also our buddy springs said to me the other day that perhaps it is a good idea to use the grain you plan on using for spawn as your BIM source, or better yet make many BIMs using various grains for maximum diversity.

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I think the seed/grain from most grasses which grow in good organic soil will be good source of lactobacillus etc.

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There is also all the members of the brassica family which have copious amounts of lactobacilli on their leaves. Perhaps canola seed, which is a brassica, would be another good choice for a source of LAB, while the fermented canola grains leftover after straining off the LAB liquid might be useful as a fungi growing supplement.

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Good idea.  I wonder how canola seed is as a spawn sub.


Thoughts?


THanks! 

P.S. I plan on posting this at your site too but not for a few weeks and I need to edit it for more general use than just focused on fungals.  I am going to include some posts of members like you and mycelio (if he's OK with that), etc.

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Here's what I propose:
(in this example I'm using LAB, but AEM, BIM mix, etc are interchangeable)

1. Ferment two different LAB pure cultures, one with 4.5% (1:1:20) and one with 8.5% (1:1:10).  LAB pure cultures are taken from same mother culture.
   
   
2. Place organic matter (rye, WBS, straw, h.poo, etc) onto a cookie sheet and place in oven at 175-250F for 24-48+ hours.  This should remove all internal moisture content from organic matter.
   
   
3. Weigh out four equal amounts of: rye, WBS, straw, h.poo.  Record weight measured.
   
   
4. Individually ferment rye, WBS, straw and h.poo in 4.5% and 8.5% pure cultures, and two control samples of distilled, boiled water. (all weights of organic matter and volumes of liquid are the same)
   
   
5. After day 7 of fermentation (low o2, 75-90F) drain off liquid and place organic matter on cookie sheet.  Place cookie sheet into oven at 175-250F for 24-48+ hours until all internal moisture is removed. (I am going to carry out many of these trials in small samples so I can test on different days)
   
   
6. Weigh samples again.  Lightest samples most likely had the greatest digestion by enzymes, acids, etc.  IMO the lightest would probably also have the greatest amount of fermentation.  Thoughts?

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Heating above the boiling point of water for one or two hours might be enough.

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More important: Wouldn't the microbes stay on and in the kernels? Then the weight difference might be very small and would only tell about substances that went into solution.
Unfortunately I have no idea on how to remove our little helpers, so only the grain would remain. I guess shaking wouldn't help.

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ABSTRACT:
The purpose of this study was to determine the optimum concentration of blackstrap molasses required in conjunction with the use of several species of homofermentative lactic acid bacteria in the fermentation of hydrolyzed cod gurry. Blackstrap molasses at a concentration of 7.5% was found to be optimum for achieving a successful lactic acid fermentation of hydrolyzed cod gurry. Among the three homofermentative lactic acid bacteria studied (Lactobacillus plantarum, Pediococcus acidilactici and Streptococcus faecium) L. plantarum resulted in the most rapid fementation. S. faecium was the least desirable bacterium on the basis of fermentation rates and extent of pH decrease after 72 h of incubation. All three organisms failed to utilize significantly the sucrose derived from molasses, even though sucrose was the most abundant carbohydrate in the molasses. P. acidilactici was unable to utilize sucrose when present as a sole carbohydrate, while L. plantarum and S. faecium decreased the pH in broth cultures with sucrose as a sole carbohydrate. These results suggest that glucose in molasses functions as a repressor of invertase formation in L. plantarum and S. faecium, and that the use of derepressed mutants should allow a significant decrease in the amount of molasses required for a satisfactory fermentation.

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