So, after a little gearing up and doing some research im finally back with new plans
,
As already said Lostmushroomforests´s replys on my experiments got me major thinking.
So before going on culturing the fleece on greater scale i definitly have to know more exactly what i actually have in culture, because of this hole productive Lactobacillus GAD Enzyme and Yeast´s contraproductive GABA/Musicmole consuming properties thing.
I know its not clear what exact Molecule is producing the Fleeces effects, but my assumption is it could be a similar Molecule to Muscimole.
And therefore it could be very possible that the Lb´s GAD and the Yeasts hunger could affect the fleeces properties in a same way then they would affect the muscimole.
This could explain the varying effects people get by consuming fleece products. I didn´t realize i´m cultivating not only the fleece but a hole symbiotic tohuwabohu until
@lostmushroomforest pointed that out (thanks again haha). If my assumtions are right the yeasts or other microorganisms could have easyly consumed nearly all actives away before i had the chance to.
Maybe this is the unconsious experience of the most fleece enjoying people out there ? this could be the key of the fleeces unreliability when it comes to consistentsy of effects.
Can you people out there who already got into the bliss of fleece effects confirm this inconsistens in effects ? (
@Mcpato ,
@EaglesNest ) as far as i read the posts in this forum it seams so.
The don likely had a very helpful simbiotic culture when you read his comparison of the effects with the effects of LSD
(Donald E Teeter - Amanita Muscaria, Herb of Immortality; Page 51)
So what i did now is big researches in the cultivation and (health)benefits of Lactobacillus.
I can recommend the researches of Lostmushroomforest in his posts in this forum
viewtopic.php?f=5&t=712&start=10#top
but there are many more
a couple of posts ago i did post a little list of research starters:
i think i´ll try simply putting lactic acid in raw ambrosia, but more promising seems letting ambrosia colonise by a lactobacillus culture or maybe letting a lactobacillus culture colonize by the fleece
or even letting them colonise simultaneously ? in which mediums could anything of that be possible ? maybe even in grape juice ? maybe with added nutrients ?
i´ll try to follow the advicements in the different lacotbacillus strains thread (first link below) when it goes into picking one of the many lactobacillus cultures.
viewtopic.php?f=25&t=758 (-> different lactobacillus strains, great thread @lostmushroomforest ! i´ll definitly post there when i have results !)
viewtopic.php?f=5&t=684&start=10 (-> amanita with raw milk -> soma by @amanitadreamer )
viewtopic.php?f=5&t=647&start=10 (-> lactic acid conversation thread by @Zopphire )
https://www.dmt-nexus.me/forum/default. ... ts&t=87052
&
https://www.dmt-nexus.me/forum/default. ... ts&t=95478 (-> similar thoughts about this topic at the dmt nexus forum)
also the book
Lactic acid bacteria microbiological and functional aspects (ISBN-13: 978-1439836774, ISBN-10: 1439836779) was very helpful. (dont kno if im allowed to post the page where people out there get a free pdf but i think thats not complicated anyway nowadays)
also i oriented on a guide Lostmushroomforest posted in one of his threads:
https://www.fivebladesbrewing.com/optim ... us-growth/
https://www.fivebladesbrewing.com/lacto ... ter-guide/
and one of the references the guy used:
http://www.milkthefunk.com/wiki/Lactobacillus
So as you can already make a sense out of the infos i used, my quest is now getting a clean culture of only the fleece and the Lactobacillus.
In the future i´ll definitly try to find good wild Lactobacillus strains like in raw milk recommended often in this forum or sauerkraut or something like this,
but for the start i´ll use a pure culture of Lactobacillus brevis i found in a hobby brewery delivery. they using it for giving the brews a more and additional sour aroma.
I´m very lucky because the lactobacillus brevis culture is the only pure lactobacillus culture they sell and this is one of the good ones when it comes to lactic acid and GAD producing properties like you can read in Lostmushroomforests research (
viewtopic.php?f=5&t=712&start=10#top)
I will likely try letting the medium colonize by the Fleece and afterwards by the lactobacillus and vis versa and off course also letting them colonize simultaneously. i think they should be able to build a working symbiosis but i will have to try it out first. this has to be done all under strict sterility because i want them to establish without the presence of yeast.
As
@Mcpato found out the fleece is able to survive under aerob and anaerob conditions, but will do better and faster with air.
My researches for the Lactobacillus result most of them favor anaerob conditions but can tolerate oxygen. ("facultative anaerobes")
Some of them produce CO2 and some don´t, Lactobacillus brevis does produce a bit CO2.
So the first thing i´ll do is a starter culture with anaerob conditions.
i built a jar with injetion port and CO2 release type of thing you´d use for brewing. just imagine the anaerob pendant of the jars some people would use for liquid culture of fungi. hehe
- anaerob jar.jpg (86.63 KiB) Viewed 199706 times
this will be pressure cook sterilized, of course without the plasic CO2 release thing (damnnn forgot how to call it in english) which will be added afterwards again under sterile conditions, while the hole gets covererd with a couple of layers micropore tape.
I thought about strong salt water to put in the CO2 release thing? (prevent microorganisms without using strong toxic solvents which might be bad for the plastic)
for the medium i thought about a glucose medium (20-30g/l tap water (we have very good quality tap water here))
and a little bit of yeast nutrients. the pH should be between 5-6 or sth like this (will have to look a again haha) for this i have maybe tartaric acid or of course acetic acid in form of 25% vinegar.
i´ll try glucose at first but future runs maybe with malt extract honey or maybe maple sirup.
- lactobacillus and yeast nutrients.jpg (85.9 KiB) Viewed 199706 times
the lactobacillus is called
Lactobacillus Brevis WLP672 and more informations can be found at the whitelab homepage.
this is the method i chose, because it will enable the possibility to succ a bit of it out with a needle and test for pH or starting a new culure or inject new nutrients with sterility.
so much about the lactobacillus part of the project,
the fleece part of the project will be a bit shorter.
at the moment i run a couple of cleaning up steps in vermiculite and brown rice fluor now under pressure cooked sterile conditions.
i tried to pick some of the black spores and inoculated a new jar under sterile conditions as possible, but thats a bit difficult because all jars with fleece are not sterile (look at my posts before) and opening a jar and collecting spores with tweezers and closing again isnt that sterile without a laminar flow hood ...
but i hope with a couple of cycles there shouldnt be much more than the fleece (need to build a still air box tho)
my plan is to build a sterile starter culture of fleece where you can take new inoculant with a needle not with tweezers which would be much more convinient.
Like some other people would do it with grain to LC or LC to LC (shroomery or mycotopia for more informations)
So i hope this hole effort will bring some more conclusions, but i think even if this all results in no noticable differences we can conclude the fleece actives are not affected by the yeast and/or the GAD, haha
BUT i am relatively optimisic it will change the experience and give more consistent results if my assumption that it is a similar molecule than ibotenic acid/muscimole are true,
BECAUSE i think it could be possible that the fleece metabolizes some (whatever) nutrients to some kind of molecule with similarities to ibotenic acid/muscimole (our active) and then the amanita mushroom metabolizes that compound to ibotenic acid/muscimole.
This would of course only be possible when the assumptions are true that the fleece lives naturaly in symbiosis with the amanita mushroom (in the amanita mushroom probably) that is still no fact so far as i know but very good possible because many people around the world get the fleece when trying to isolate it from amanita.
Another indication for my assumtion about the impact of GAD to our infamous possible Muscimole similar Molecule is the fact(?) that at first there was only known the GAD would impact Glutamate and turn it to GABA, then (as described in the trent austin paper) the GAD can turn ibotenic acid to muscimole (pls correct me if i misinterpret anything of that) was found out, and now i assume why shouldnt the GAD be able to decarboxylize any or at least some more molecules with the glutamate ground structure?
- glutamate to GABA.PNG (22.5 KiB) Viewed 199706 times
when you look at the structure of ibotenic acid and muscimole it does the same thing, it just removes the carboxygroup (->decarboxylase)
- ibotenic acid to muscimole.PNG (53.07 KiB) Viewed 199706 times
so ibotenic acid and muscimole nearly have the Glutamate/GABA ground structure in it, but there are well known synthetic compounds out there which are even closer to GABA then Muscimole. Look at Pregabalin (lyrica) Gabapentin and phenibut, their theoretical carboxylated counterparts would definitly get decarboxylated by the GAD, too.
- structure similarities.PNG (94.97 KiB) Viewed 199706 times
(no idea what this bicuculline and saclofen things have to do on this pic but the other molecules are showing good what i want to say)
now i think it gets a bit clearer what i´m trying to explain. My claim is that the molecule produced by the fleece is a molecule which has a similar ground skeletal structure than all these glutamate/GABA like compounds.
In its raw form it has a carboxy group (carboxy groups give the molecule sour properties (proton donor))
and in its 'reacted' form the carboxy group is removed (decarboxylated -> decarboxylase (GAD/Glutamate Decarboxylase))
This is the reason of the don claiming to heat it up to decarboxylize it, because it actually works in the same way as with ibotenic acid/muscimole, but enough threads in this forum show that it works bad/work better with other methods like with GAD.
(in the future i´ll try to create a more fancy pic of the chemical background of my claim)
This could be another reason why some people (for example
@Mcpato claims he has better results in not heating/decarboxylating the ambrosia) because maybe he/you have/had a culture which was already decarboxylated well before heating, which could be contraproductive in this case.
Or some people get poor effects with the fleece heated or pre heated, because there are already many goodies-consuming yeasts in their symbiotic culture,
of course they have poor results no matter what they are doing with it, and than they are doubting this hole fleece-thing!
of course this is all very theoretical and many tests must be done to prove my claims,
but i hope you guys are willing to go through this journey with me!
it could be a huge step in understanding the mushroom and the fleece,
imagine, this hole sterility and yeast nutrients type of thing sounds a bit scaring difficult and very much erfort and so on, (at least for the amateur home DIY-kind of person like me) but if there results an easy-to-handle and more-consistent-in-effect type of SCOMBY with the fleece, it would be a huge step forward! and of course there will be much easyer ways to create a good SCOMBY, with raw milk for example and i think the strict sterility isn´t as important as it looks in this post, but at the first runs i have to be sure of course! and afterwards there will come the big jars again
i´ll post back soon with my progression going on!
thanks for reading, your attention and thanks to all participants in this thread before and in my timeline so far!