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 InTheRainySeason
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 Re: Grain spawn without pressure cooking [Re: Anand]
#8936117 - 09/15/08 08:27 PM (3 years, 8 months ago) |
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Should work well for P Cyan. I read some great things in quickpick's LAB thread about how it breaks down materials and makes them more available as nutrients. Wood lovers are already so tough, these microbes will probably make growing them even easier.
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 Anand
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Well the part with the EM liquid is not fruiting, the other is.... 
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InTheRainySeason
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Re: Grain spawn without pressure cooking [Re: Anand]
#9028896 - 10/04/08 01:32 PM (3 years, 7 months ago) |
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That's a bummer. I found some notes I thought were interesting...
"Large numbers of spores in a concentrated area can overwhelm just about any fungicide. Growers have even inoculated peroxided agar with a large swipe of spores and had a few survive to germinate. The other possibilities exist as well, of course.
RR "
"None that I know of. Most fungicides kill all spores, but allow mycelium to grow untouched. Thiophanate-methyl is approved by the FDA for use on mushroom crops and I ran some tests on it a few years ago. It's the active ingredient in Banrot 40WP. You can read up on it here. http://www.epa.gov/oppsrrd1/reregistration/tm/tmsummary.htm
RR "
So I'm thinking this ferment method would work with spores if you used a lot of spores. Like a dark syringe... An idea I have is to print the spores on blades of grass and use this in the ferment treated substrate. Placing the blades in a group stacked on each other on top of the substrate when the jar is only half full of substrate, then fill in the rest. I think any contam spores would be in such a small amount that the mushroom spores would overwhelm them quickly.
Unfortunately I made my liquid culture without sterile procedures, so I may be cultivating mold which I hope is really mushroom mycelium. I used all my spores up. But from the tests I've done, I know that it works well to kill foreign spores, so at least that.
Roger Rabbit also mentioned rhododendron as a fungicide that'll prevent contam spores from germinating while allowing the mushroom mycelium to grow. There was an old topic with no follow up, where they discussed using oregano oil and other herb oils as a fungicide treatment for substrates. I think this would work much like the ferment method. At certain concentrations, oregano oil will lose its effectiveness at fighting mold spores and other contamination after a few weeks. So I think it would work well with a concentrated spore solution or mushroom stems....
Oregano seems great because it's so hardy and easy to grow. I also live in a perfect climate for it.
Edited by InTheRainySeason (10/04/08 01:35 PM)
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acromonium
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I had a thought for outdoor grows. Instead of ant traps and all the bad karma associated with that use ground up habanero. ants wont eat it, dogs wont dig it. i dont think the active ingredient will interfere with mycelium, prolly just get consumed.
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CheeWiz
Registered: 09/28/08
Posts: 265
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Re: Grain spawn without pressure cooking [Re: spacel0rd]
#9041767 - 10/07/08 10:29 AM (3 years, 7 months ago) |
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What you have here is the old mycology trick of using a low PH to inhibit the growth of bacteria, plus some form of symbiotic relationship between the mushroom your trying to grow and whatever the bacteria that's producing the lactic acid. What would be really great is if you could isolate the symbiotic bacteria. The problem with natural endogenous bacteria and their endo spores is that they do very from plant to plant, local to local and you may have been lucky in that your grain has the right strains of bacteria. That's why I say isolating it would be helpful information for us to have so we could better under what's going on.
The growing of mushrooms goes back 1000's of years. Well beyond leister and others or PC's. Our ancestor learned at first by seeing what was going on in nature then trial & err and some just plain good luck. It kind of makes one wonder is that before there were PC's if soon thing like this may have been used in growing their base spawn and for the all most religious use of rye grain in preparing spawn. Many times its regular people that come up with the most useful discovers and not some eggheads that's all too eager to claim credit.
What I'm working with now is clean washed straw that has been inoculated with Bacillus Subtilis, Natto grown on soy beans. There are many forms Bacillus Subtilias found around the world and most are pro biotic in many whys and is one of the bacteria that survives digestion in equines and found in their dung. But more on that at a later time as I'm in the early stages. Hipster
Edited by CheeWiz (10/07/08 10:32 AM)
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InTheRainySeason
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Re: Grain spawn without pressure cooking [Re: CheeWiz]
#9043767 - 10/07/08 04:39 PM (3 years, 7 months ago) |
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That's awesome. Maybe one of you will isolate the perfect bacteria with the most anti-contam qualities.
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Mycelio
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@Hipster
Bacillus subtilis might be useful for fermenting substrates, which are rich on nitrogen, like mixtures of straw and beans. This would be interesting for species like agaricus bisporus. My previous experiments all ended with the foul odor of butyric acid...
Please keep us updated about your experiments.
@inTheRainySeason:
Good find about the use of masses of spores, though it may be much easier to germinate a few spores on straw or fermented straw and then inoculate fermented grain with active mycelium.
The effect, that some species of mushrooms won't colonize and spores won't germinate, may be caused by the low PH, rather than by antibiotics.
Just found a German website, explaining microbial life in sourdough, which is the same as in our fermenting grain. They talk about three groups of species. Group one and two are lactobacillus, which break down starch into sugar, where group one is the majority and more effective. Group one produces only lactic acid from sugar, group two does too, but also creates acetic acid and some CO2. Group three consists of acid tolerant yeasts, which produce alcohol and CO2 from sugar. These yeasts are not the ones, that may grow on the surface, if the fermenting grain gets too much fresh air.
A few common species in sourdough:
Group 1:
- Lactobacillus plantarum
- Lactobacillus casei
- Lactobacillus delbrueckii
- Lactobacillus leichmannii
Group 2:
- Lactobacillus brevis
- Lactobacillus fermenti
- Lactobacillus pastorianus
- Lactobacillus buechneri
Group 3:
- Saccharomyces cerevisiae
- Pichia saitoi
- Candida crusei
- Torulopsis holmii
Then some German scientists found lactobacillus reuteri in sourdough and examined its antibiotic product, called reutericyclin. This substance is reliable for killing other types of bacteria.
Carsten
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spacel0rd
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Re: Grain spawn without pressure cooking [Re: Mycelio]
#9046044 - 10/08/08 12:03 AM (3 years, 7 months ago) |
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@ Mycelio:
So germinating spores on fermented straw actually works? :O
That's pretty cool. 
Unfortunately my second straw experiment failed again. This time I put the straw in jar to have a better anaerobic condition than in a bucket. I even had some grain in that jar as well, because I felt grain was doing better for fermentation. But still, it got really ugly. Opened it after a few days and again: slimy and stinky.
So I advise everybody to rather use straw pellets as Carsten did. I will do so soon.
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Mycelio
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Re: Grain spawn without pressure cooking [Re: spacel0rd]
#9046497 - 10/08/08 05:10 AM (3 years, 7 months ago) |
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Hi Spacelord,
I once tried cube spores on straw. The straw started fermenting, but mycelium showed up after three weeks. So it works, but you need patience. It should be faster, if the spores germinated already. You can fill a syringe with straw tea (a few pellets boiled in water for 5 - 10 minutes) and some spores. Germination will happen after two days. Then shake and inoculate with single drops or more. So far I never had contams this way, even with inoculating sterile grain.
Carsten
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Burn it up
Soma
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Re: Grain spawn without pressure cooking [Re: spacel0rd]
#9080125 - 10/15/08 07:15 AM (3 years, 7 months ago) |
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Great post, man.
Very interesting
--------------------
-Purple Placebo-
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baltazar
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Re: Grain spawn without pressure cooking [Re: Burn it up]
#9236595 - 11/13/08 09:29 AM (3 years, 6 months ago) |
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Can the grain, fermented using this method, be inoculated with LC?
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Mycelio
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Re: Grain spawn without pressure cooking [Re: baltazar]
#9236682 - 11/13/08 09:50 AM (3 years, 6 months ago) |
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Hard to say without knowing which species... Better try mushroom stems or put a spoonful of colonized substrate on top.
Carsten
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Anand
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Re: Grain spawn without pressure cooking [Re: Anand]
#9536444 - 01/03/09 05:53 AM (3 years, 4 months ago) |
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Quote:
Anand said:
Well the part with the EM liquid is not fruiting, the other is....
Looks like all the myc is gone in the part that i used the EM on, only the EM myc is growing
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Mycelio
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Re: Grain spawn without pressure cooking [Re: Anand]
#9536619 - 01/03/09 06:59 AM (3 years, 4 months ago) |
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Too bad! Does that mean you have started a LC with bokashi?
Carsten
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goose2r
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Re: Grain spawn without pressure cooking [Re: Anand]
#9536967 - 01/03/09 08:52 AM (3 years, 4 months ago) |
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Quote:
Anand said:
Quote:
Anand said:
Well the part with the EM liquid is not fruiting, the other is....
Looks like all the myc is gone in the part that i used the EM on, only the EM myc is growing
EM doesn't have myc, it has no fungi besides sugar yeast. The original formulation of EM used to have so-called "ray" fungi but no longer. You maybe noticing the yeast output.
-goose2r
Edited by goose2r (01/03/09 09:12 AM)
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InTheRainySeason
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Re: Grain spawn without pressure cooking [Re: goose2r]
#9682704 - 01/26/09 12:55 PM (3 years, 4 months ago) |
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I'm guessing this is the particular species that's blocking contamination. http://www.ncbi.nlm.nih.gov/pubmed/12561574
And they sell starter culture of it http://www.bodyecology.com/07/08/16/lactobacillus_plantarum_benefits.php
Probably wouldn't be all that hard to isolate now that I know which one does it. There's probably many species that do it but that one seems like the best.
Isolating it might have even more contam resistant properties. And it would probably speed the process up a lot.
"Actively growing Lactobacillus plantarum CGMCC 1.1856 cells totally inhibited the germination of mold spores. Cell-free supernatant broth from the fermentation of Lactobacillus plantarum could not destroy the viability of mold spore. While the pH of the culture broth and supernatant were about 4.0, and the acidification of non-fermented broth to pH 4.0 with lactic acid could not cause a similar inhibition on spore germination. This experiment discounted the possibility that the inhibitory effect on mold growth was due to the lactic acid produced by Lactobacillus, and suggested that the effect was because of both low pH and microbial competition."
Nice to know that it's good for us. I've got some wheat fermenting, and an oat/brown rice/wheat mixture fermenting as well. I'm going to try to see if I can get the contam resistance into the rice. If I could do that it'd be really nice. Plantarum doesn't seem picky about its food so it should work. I've got a lot of mycelium to work with now. It seems that coating the stuff won't work and that it needs to be fermented with the lactobacillus so that there's a lot of cells in the spawn. I'm also going to inoculate a sourdough mixture. But it has a cup of brown rice powder along with a cup of wheat flour. I'm going to use ferment water to speed the process up when I mix it, and to make sure there's more lactobacillus in it, then when it's mixed up I'm going to let it colonize the dough for an additional 8 days.
I'm going to try various types of leaves to ferment like kimchi. I'll start with devils ivy since I have lots of that and it'll hold moisture well.
Edited by InTheRainySeason (01/26/09 12:55 PM)
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Mycelio
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Lactobacillus plantarum seems to be common. I expect it to occur in fermenting grain anyway.
Should start some sauerkraut experiments myself, though I'll use some vegetables.
Good luck and keep us updated,
Carsten
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goose2r
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Re: Grain spawn without pressure cooking [Re: Mycelio]
#9684981 - 01/26/09 07:01 PM (3 years, 4 months ago) |
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Quote:
InTheRainySeason said:
I'm guessing this is the particular species that's blocking contamination. http://www.ncbi.nlm.nih.gov/pubmed/12561574
And they sell starter culture of it http://www.bodyecology.com/07/08/16/lactobacillus_plantarum_benefits.php
All/nearly all species of Lactobacillus spp. will hinder microbes which can not live in the environment colonized and fermented by Lactobacillus spp. When using microbes you want to use a synergistic consortium so they benefit each other. A mono-culture is very rarely found in Nature. L.plantarum is one of the most common species but others should offer similar benefits they just are not as well studied.
For example, EM is a synergistic consortium of sugar yeast, PnSB and LAB. Some manufactures like SCD world also include Bacillus spp. and even Bifidobacterium spp. (which is mainly for human/animal benefit when consumed). In EM the yeast ferments with the molasses, as does the LAB. The LAB feeds upon chemicals released during the yeast's fermentation process and PnSB feeds upon dead yeast cells. PnSB also feeds upon dead LAB and other chemicals released by LAB.
If you want to use a pure culture of Lactobacillus spp. you can indeed isolate and culture it at home for very cheap. To isolate L.plantarum would be hard as you need to do serial dilutions, plate and identify, then isolate again and culture in enrichment broth.
Here is a thread a few pages back I found the other day., directions to isolate and culture Lactobacillus spp.:
http://www.shroomery.org/forums/showflat.php/Cat/0/Number/8910533/an/0/page/5
Oh yea, you should add 1% calcium carbonate powder in the enrichment broth (the molasses/water mix) when culturing the LAB spp. LAB prefers a acid/neutral pH of around 5-6.5. That thread is good but there are tweaks which would make the isolation more trustworthy (when not verifying with a microscope). The biggest difference I would do is serial dilutions into fresh enrichment broth. I would do 2-3 serial dilutions. By then the culture should be nearly/totally all LAB.
-goose2r
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InTheRainySeason
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Re: Grain spawn without pressure cooking [Re: goose2r]
#9724120 - 02/02/09 04:23 PM (3 years, 3 months ago) |
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Mycelio, you mentioned you were going to try fermented brown rice, did it work? I tried it and it contaminated, maybe cause it was too wet since it was very wet. I'm going to try fermenting it for 2 weeks instead of one this time and make sure it's not so wet.
I also had wheat flour in the mixture, and oats. That could've screwed it up. And I'm wondering if boiling grain is necessary so that it kills the contam spores before they can germinate when the grains are submerged, since I don't know if the beneficial bacteria like l plantarum would kill the already germinated mold.
I'm looking forward to isolating the lactobacillus plantarum because I know that will work, but it's going to probably need to go through the natural process of pH lowering before I can add the isolated bacteria effectively.
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Mycelio
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I do fermentation with grains covered by water, so too wet can't be a problem. Too little water and too much air favors contams. Try filling a jar half with grain and then up to three quarters with water.
Wheat flour should ferment fine, as in sourdough, but the flour will not make a good mushroom substrate, better use whole kernels.
And no, I didn't try rice yet, also oats. I boil grains only to prevent germination of the plants, nothing else. After cooking I add the water from soaking to inoculate with lactobacillus species. If you want to inoculate with L. plantarum, you should boil, adjust the PH with citric acid and add L. plantarum immediately, otherwise the natural microbes will have colonized everything.
Good luck,
Carsten
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