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Offlinekrypto2000
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Can psilocin be produced in a LC ala claviceps?
    #23826775 - 11/12/16 06:12 PM (7 years, 2 months ago)

I have not heard of this being done but then they are illegal so maybe it just hasn't been researched.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: krypto2000]
    #23832971 - 11/14/16 05:09 PM (7 years, 2 months ago)

What exactly are you asking? Claviceps does not produce pscilocin, are you interested in extracting psilocin/ psilocybin from an lc? Is that what your thinking of doing


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Offlinekrypto2000
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Re: Can psilocin be produced in a LC ala claviceps? [Re: KThunderland]
    #23833532 - 11/14/16 07:37 PM (7 years, 2 months ago)

Yes, I'm wondering if you can select for and mutate a strain or culture of psilocybe to produce worthwhile levels of alkaloids in a LC. If you could get 1g/l it would be a better option than actually growing mushrooms.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: krypto2000]
    #23834597 - 11/15/16 07:19 AM (7 years, 2 months ago)

You might have something there, ill try yo find anything that i can that could be pertinent to this, rather than shoot it down we can "think tank" this, i have heard of people eating cakes and tripping, 1g per liter is far more than worthwhile:) hiw do you plan on handling ectraction in the end? Isnt not a canidate for a/b extraction, so maybe a tincture so some sort (maybe salted to getvtge alcohol and water to seperate, my chemistry is rather rusty im gonna look into the areas where i think there wiuld be issues.)


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Re: Can psilocin be produced in a LC ala claviceps? [Re: KThunderland]
    #23834627 - 11/15/16 07:43 AM (7 years, 2 months ago)

https://www.shroomery.org/forums/showflat.php/Number/515430#515430

I always thought this would be fun to try.

I'm not sure if mutation would need to be done. I suppose w ur target goal mutation could help if the target traits were aquired..

Do it!


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Offlinekrypto2000
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Re: Can psilocin be produced in a LC ala claviceps? [Re: Raven44]
    #23835153 - 11/15/16 11:22 AM (7 years, 2 months ago)

I guess if they did it as described in that link then it is very much so viable. The one thing I question is that they seemed to only extract the biomass and discarded the nutrient broth. I find it hard to believe that nutrient broth was not absolutely loaded with psilocin if the biomass itself was, both alkaloids are water soluble enough that I would not discard it like that, especially psilocybin.

Extraction wise you actually can do an a/b just fine. I did it with water, NaOH, DCM, and GAA. 10mg of the extract is a good dose, at least equivalent to a gram of cubensis if not more I would say. The extract itself is a golden syrupy like oil which will occasionally crystalize in the air (or at least in a vacuum anyway). I would imagine it's at least 50% pure and you can get it pure with a simple recrystalization I would think, I just haven't had enough to make it worth trying. I think this is the future guys. I'm going to be expiramenting with this from now on. It might be awhile to find a suitable strain and get the process down, I don't know, but I would be surprised if it did not work quite honestly.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: krypto2000]
    #23835189 - 11/15/16 11:32 AM (7 years, 2 months ago)

Interesting thought, about the water being saturated with target alks

Maybe that's where harvest time comes into play. One day of fermentation too long resulted in a drastic alk reduction link says. I'm unsure of why that happens


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Offlinekrypto2000
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Re: Can psilocin be produced in a LC ala claviceps? [Re: Raven44]
    #23835481 - 11/15/16 01:16 PM (7 years, 2 months ago)

I'm sure that's what happened, just think about it. Psilocybin especially is very water solube so it seems unreasonable that it would *not* leech out into the broth. In claviceps fermentations I've read that ~80% of your alkaloids tend to be in the broth while only ~20% is intracellular, those that are intracellular tend to be more non polar as well, both psilocin and psilocybin being rather polar I would not be surprised if even less than 20% is in the mycilial mass.

This is not exactly an apples to apples comparison but neither is it apples to oranges either. To put this in perspective both ergot slcerotia and psilocybin mushrooms are about 1% alkaloids by dry weight. A wild strain of claviceps will maybe produce 50mcg/l with little or no slection. With selection and mutation this can easily be brought to over 1g/liter under standard fermentation times. 2.5g/l is about the highest I've read of and 8.5g/l is the highest I've read with a prolongued fermentation (60 days as opposed to ~10 and 6 medium transfers as opposed to ~2).

Now if you compare this directly to the above reference fermenting a high yield strain of claviceps in a 55 gallon drum would yield about 208g of ergolines. If you extracted just the biomass and threw out the broth you would be getting ~40g of alkaloids which is about 5x more than that of the psilocybe fermentation. This tells me there is *vast* room for improvement. I mean for one just look at it from a nutritional perspective. If you put in 100g of carbohydrates *something* is happening to them. Either it is being turned into vegetative growth or some type of metabolite. All that carbon went somewhere.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: krypto2000]
    #23835703 - 11/15/16 02:44 PM (7 years, 2 months ago)

I hear u, Def room for improvement but a good lil link lol

Good luck :smile:


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Offlinekrypto2000
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Re: Can psilocin be produced in a LC ala claviceps? [Re: Raven44]
    #23835836 - 11/15/16 03:28 PM (7 years, 2 months ago)

Okay, so reading that link more I've noticed two things. First off they do get a 1% yield however this seems to be only based upon the mycilial mass, not 1% of the glucose added. If they had a 1% yield per glucose they would have gotten about a 20g yield. Odds are that is exactly what they got however they just negelected to extract the nutrient broth, I really can't fathom why other than short sighted thinkong from previous experience with mushrooms.

That brings us to the 2nd point though which is the glucose content itself is extremely low relative to the volume of the container. Claviceps fermentation for instance uses up to, iirc, 50 or perhaps 100g of glucose *per liter*. It has also been shown that a large portion of that glucose can be substituted for standard table salt. I believe the reason this is is the high glucose acts not only as a carbon soirce but it also changes the osmotic pressure allowing nutrient update. Perhaps there's something else to it and that osmotic pressure also signals the cells to limit their vegetative growth. If there is no room to grow for instance it stands to reason the cells would somehow know this and focus their resources elsewhere, perhaps on secondary metabolite production. That last part is pure speculation so take it as you will.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: krypto2000]
    #23836275 - 11/15/16 05:21 PM (7 years, 2 months ago)

from an old thread
Quote:

blackout said:

The post about "soft agar" was calling it the "Brack & Kobel" method. Have a read of this
https://www.shroomery.org/forums/showflat.php/Number/7680529#7680529

If you do not want to bother growing fruitbodies I would recommend growing stones/sclerotia. Minimal equipment is needed and it is a lot easier to see if they are contaminated or not. Some here have harvested and tripped on stones that were only grown for around a month, if you are in a hurry, but they should increase in yeild and potency if left the usual 3-6 months.

I want to give growing stones on soft agar another go, it would be extremely easy to harvest and suits my own setup.




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Offlinekrypto2000
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Re: Can psilocin be produced in a LC ala claviceps? [Re: blackout]
    #23836322 - 11/15/16 05:37 PM (7 years, 2 months ago)

Hmm.. thanks. It seems that is basically half way to what I'm trying to do. It says in the article that LC works just fine, the main limitation is just aeration. But then how much does it cost to buy a simple air pump, 20$? And then how much does agar cost? I think for a few batches the author is right on, but after even just two or three you're paying more for agar than you would be for an air pump.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: krypto2000]
    #23836353 - 11/15/16 05:46 PM (7 years, 2 months ago)

Quote:

krypto2000 said:
It says in the article that LC works just fine, the main limitation is just aeration.



Can you quote the bits where you reckon it says its "fine", what I take from that thread is that LC is highly inadvisable and that the key is to use soft agar. Agar is expensive stuff, but it is using far less than usual.

I keep planning on trying this out again, I tried before but it contaminated.


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Offlinekrypto2000
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Re: Can psilocin be produced in a LC ala claviceps? [Re: blackout]
    #23836671 - 11/15/16 07:38 PM (7 years, 2 months ago)

This is what I was referring to:

Quote:

The beauty of a soft agar medium is that it ensures that the growing mycelium doesn’t sink to the bottom of the cultivation container where it can not breathe and stops growing.

Problems of liquid cultures

Lack of oxygen is a major problem in liquid cultures. Paul Stamets devotes a full chapter on liquid cultures in his cultivation bible Growing Gourmet and Medicinal Mushrooms. In there mycelia are grown out on petri dishes of solid agar, fragmented in an Eberbach blender, incubated in Erlenmeyer flasks and aerated by use of a magnetic stirbar and a stirplate which is put at 100-200 rpm. All this use of expensive equipment and the extremely contamination-sensitive practice of culture stirring becomes obsolete when one only adds some agar gelling agent to the medium, so that the mycelium stays at the surface.




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Re: Can psilocin be produced in a LC ala claviceps? [Re: krypto2000]
    #23840853 - 11/17/16 07:30 AM (7 years, 2 months ago)

Quote:

krypto2000 said:
This is what I was referring to:

Quote:

The beauty of a soft agar medium is that it ensures that the growing mycelium doesn’t sink to the bottom of the cultivation container where it can not breathe and stops growing.

Problems of liquid cultures

Lack of oxygen is a major problem in liquid cultures. Paul Stamets devotes a full chapter on liquid cultures in his cultivation bible Growing Gourmet and Medicinal Mushrooms. In there mycelia are grown out on petri dishes of solid agar, fragmented in an Eberbach blender, incubated in Erlenmeyer flasks and aerated by use of a magnetic stirbar and a stirplate which is put at 100-200 rpm. All this use of expensive equipment and the extremely contamination-sensitive practice of culture stirring becomes obsolete when one only adds some agar gelling agent to the medium, so that the mycelium stays at the surface.







i'm a "professional" mushroom grower and I wanted to drink the LC coolaid, mostly because it would save a whole shitload of time inoculating.  However, out of maybe 12 attempts I only got one contam free batch.  No idea where the yeast gets in, but I'd say it's quite difficult to do properly without a very clean, cat 100 clean room and perfect sterile technique.

unless you're dead set on this experiment, stones seem to be a much cheaper way to go.

Proper incubation flasks with stirring and aeration are $1000 minimum.  luckily you can pick them up second hand for about that, in larger sizes over a gallon if you're really that into it.


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Offlinekrypto2000
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Re: Can psilocin be produced in a LC ala claviceps? [Re: drake89]
    #23841006 - 11/17/16 09:14 AM (7 years, 2 months ago)

Are you using a flow hood? I get contaminated LC often but not that often and it's always down to my technique or a contaminated starting culture. If you can make sterile grain you can make a sterile LC.

As for a fermemtation flask I'm sure a modern commercial fermentation vessel cost 1000$ but that's not the only options out there. A 1 liter erlenmyer flask and polyfill costs ~15$. A cheap consumer grade magnetic stirrer costs ~30$, and then a stir bar maybe 5$ more. So 50$ compared to 1000$.

You could also go with a chinese lab grade bioreactor which is ~100$, but then you would still need either a stir plate or an aeration device and that is overkill for this either way.

My idea is to take a bunch of pint, quart, or half gallon canning jars and use those in conjunction with a ~30$ aquarium air pump. I have not done it yet but on paper it should work. Get some tubing that can be sterilized to make an air inlet port and stuff it with polyfill. Use an adaptor to connect some tubing from the air pump to the inlet hose, perhaps with an adaptor if the hose is not sturdy enough, and there you go. You might need a variac to control the speed of the pump, which is actually the most expensive part at around ~60$. I suspect it would cost maybe 100$ in parts, and trial and error of parts might add more, to get a single vessel up and running. After that though it's easy to expand as a single pump can probably aerate 10s of jars, maybe even 100.

It's pretty comparable in costs to building a few monotubs if you ask me. If you do this method you are doing so with the intention of extracting it so you would already have ~100-200 in equipment and chemicals for that, if you do that what's another 100$? You also habe to consider a crystal or highly concentrated syrup is a more valuable end product than raw mushrooms so you should expect a comparably higher cost of production.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: krypto2000]
    #23841081 - 11/17/16 09:44 AM (7 years, 2 months ago)

The sterilizing of the 55 gal stainless steel drum which costs 5-6 bills is made to seem difficult

However, people use drums and sterilize stuff in custom modified 55 gal drums. They simply use a low psi and elongate pc cycle.

Use of ozonated water rather than bleach or alcohol would be mine method of madness w the drum. Just saying.

If there's a will there's a way. It also costs to play.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: Raven44]
    #23841091 - 11/17/16 09:49 AM (7 years, 2 months ago)

This is basically a thread talking about a psilocybe bioreactor.

ScleroTia are Farr less interesting compared to a psilocybe bioreactor imo


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Re: Can psilocin be produced in a LC ala claviceps? [Re: Raven44]
    #23841372 - 11/17/16 11:27 AM (7 years, 2 months ago)

look up "carlsberg flask"

also the whole first page of this thread
http://discussions.probrewer.com/showthread.php?55700-Laboratory-propegation-and-Brewery-propegation-of-yeast-(SOPs)

if you wanted you could insert a few water heater elements in there and a pressure cooker type thing at the top and sterilize inside the propagator

in the fuel ethanol industry they pump their bioreactor/fermentation full of antibiotics so that they don't have to worry about sterile technique as much. though this is a very very horrible practice fuel ethanol has greased the fingers of politicians so that they can dump out a enormous amount of antibiotics into water supplies because we really need to turn corn into gas to satisfy liberal environmental hypocrisy


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Re: Can psilocin be produced in a LC ala claviceps? [Re: bodhisatta]
    #23841449 - 11/17/16 11:47 AM (7 years, 2 months ago)

i have the aquarium pump and inline hepa filters as well.  i don't really trust a $5 filter though, so maybe you can use like 4 or 5 inline.

do I have a hepa filter?  yeah and I don't know how the bugs got in.  I only make 1000 sawdust bags a week :wink:.  can't be asked to make my own spawn any more...


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