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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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OfflineKThunderland
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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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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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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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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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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.

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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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Re: Can psilocin be produced in a LC ala claviceps? [Re: drake89]
    #23841452 - 11/17/16 11:50 AM (7 years, 2 months ago)

I trusted a $5 hepa the problem is the line after the filter has to be sterile or you're blowing sterile air into a dirty tube


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Re: Can psilocin be produced in a LC ala claviceps? [Re: bodhisatta]
    #23841746 - 11/17/16 01:21 PM (7 years, 2 months ago)

Yeah, that's what I'm saying. Just like a regular jar lid/filter except instead of stuffing the filter in the hole you will stick some tubing through the hole, glue the tubing to the lid, and then stuff the tubing with polyfill, that way the whole thing gets sterilized and no nasties should get in. I've yet to test it like I said, if I do this I will at least make another one that I did not inoculate just so I can be sure the filter is actually filtering properly.

Raven, a 55 gallon drum is analogous to starting off your mushroom hobby by building a 10x10ft outdoor greenhouse, kind of overkill. I think a 7g/55 gallon yield is pretty poor, but even so 7 grams is a ton of doses so even if you get poor yield it would be worthwhile to do a single liter. I'm convinced right off that if you were to double the carbohydrates alone you would double your yield, so that's 250mg/gallon. Far from great, but that's a good 20-25 doses at least, much more comparable to standard size monotub. I did not mention it earlier, but cost and materials aside if you are extracting pure psilocin then extracting it from an aqueous solution is much preferably to fruit bodies or mycilium, half the work is practically already done for you.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: krypto2000]
    #23842149 - 11/17/16 03:13 PM (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.







Stamets was growing LC to inoculate grains. He was not producing psilocybin. I have read plenty of posts about it, in the link I gave you will see them saying how harvest timing is critical with liquid, but not with soft agar.

I have read that the "truffle brothers" sclerotia production has the capacity for growing 18 tons per year, another farm procare had the same capacity (truffle brothers say 18,000 tons but I am pretty sure they mean kilos). Growing was and is legal in many countries yet I have never heard of any using LC for production. In the Netherlands dried mushrooms were supposedly illegal, yet they were openly on sale there many years ago, so selling dried "mushroom powder" would have been a viable option -if it was worthwhile.

You cannot visually tell if LC is contaminated or not, at least with the soft agar you can have a better guess, with stones it is far more obvious.

There is also the question if the LC will even be active, many reckon it will not be, or have far less than you might hope for.

If you do a search for threads where I mention the word "keg" you might find my ideas for bulk LC with simple & cheap equipment.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: blackout]
    #23842569 - 11/17/16 04:57 PM (7 years, 2 months ago)

Those guys who make the atlantis truffles or w/e they are in amsterdam use spawn bags inoculated with LC. I saw a documentary or mini doc on it some years back.

Otherwise what you say is true, or likely so in the case of speculation. I would imagine that most LC in fact would *not* be active, which is why it took 55 gallons just to get 7 grams. However the same is true of claviceps/ergot and yet industrially we see yields over over 2% which is twice that found in nature in the sclerotia. I'm not here because I think it's easy though, I'm here because I think it's possible. I believe the hardest part will by far be finding a strain that produces in LC as well as dialing in the nutrients and evironmental conditions to favor alkaloid production.

As to why you do not see psilocin sold in amsterdam in the past I think comes down to economics, knowledge, and by extension feasibility. Mushroom growing itself is a rather niche hobby and there is not much competition in that market, legal or not. So to the first point there is simply no demand because by and large people do not know what they don't have. It's the same reason we could produce hash oil 30 years ago to the same standards of today yet you're just now seeing it commercially and by extension it's grossly increased in popularity.

Now on top of the economics you have to first not only know that you can viably extract it but you also have to know that you can do so not only at minimal cost to you, but minimal extra costs to the consumer. As a new product people will likely be wary first off and if it costs 2x as much as mushrooms it's likely to not have a wide adoption. If it's only 20% more or even the same price then people will likely prefer it over whole mushrooms. This also ties in to why I want to make a LC.

Extracting 1g of psilocin from 1 liter of liquid is far more feasible and worthwhile than extracting it from 100g of mushrooms. I mean I would probably need a good 4 liters or so of water or more to just extract the 100g, not to mention the pain of filtering off all of the mushrooms gunk. With a LC, if it pans out, you are left with a perfect candidate for extraction. It practically begs to be extracted. With mushrooms it's not a difficult extraction, but it's a chore at the very least. I should say that I have only been using extract/crude psilocin since I first successfully did it and I have no desire to ever eat fruit bodies again having ingested it this way, that's my main reason for considering this is not only would it be novel to have but it would potentially save me a lot of time, effort, and solvents in the long run.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: krypto2000]
    #23843142 - 11/17/16 07:19 PM (7 years, 2 months ago)

In essence hear we are still talking about extraction from myc, would it not be easier to grow on something that the mycelium can be easily freed from, like corn, then agitate it with sterile water and use the resultant myc laden water to proceed with whatever extraction. I cpuld be way off but i experiance more biomass in less time on my grains than i do in an lc almost every time, to the point that ive mostly stoped with lcs.  Another problem us going to be the extraction , whats the plan there?


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Re: Can psilocin be produced in a LC ala claviceps? [Re: KThunderland]
    #23843312 - 11/17/16 08:12 PM (7 years, 2 months ago)

If you're growing it on grain you then have the biosmass + leftover grain to extract from, you might as well just grow mushrooms then. I am wondering just how much biomass there will be in the LC though, if it's comparable to the weight of the mushrooms themselves then you might have to do multiple extractions on the mycilium anyway which would largely defeat the purpose. The extraction itself is not difficult, it's just a basic a/b and then a recrystalization should get you relatively pure psilocin.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: krypto2000]
    #23844226 - 11/18/16 06:06 AM (7 years, 2 months ago)

The mycellium could be removed via agitatoin esaily from corn (normally not tye best grain byt it has an advantage here) , either way though the mushroom is still gonna always be the better option just exploring ideas


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Re: Can psilocin be produced in a LC ala claviceps? [Re: KThunderland]
    #23844230 - 11/18/16 06:08 AM (7 years, 2 months ago)

A/b wont work its a zwitterion, and alcohol tincture ia the best way with mush goodies, but then ypu have to deal with the water, which is miscible with the everclear thats used. To date i havent found a workable a/b extraction even with vwry very careful ph controls


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Re: Can psilocin be produced in a LC ala claviceps? [Re: KThunderland]
    #23844268 - 11/18/16 06:36 AM (7 years, 2 months ago)

Being a zwitterion does not mean an a/b will not work, it just means it has an internal salt, but it can still be influenced by ph and form a salt on its freebase. I've done it successfully.

https://www.shroomery.org/forums/showflat.php/Number/23420498#23420498.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: krypto2000]
    #23844452 - 11/18/16 08:02 AM (7 years, 2 months ago)

What kinda yeild are you pulling? Is that yeild worth it wuth the lower content of goodies that would be on the lc? Whats the truck i havent gotten anything close to a good yeild with a/b of shrooms and ive a/b everything lol but it you have a tek taht works then thats always gonna be the better way. Im still here to see where this go to advance the theory but it think even waiting for knots would really boost the end yeild, something that we cant do in an lc. Mind you im not trying in anyway yo shoot down the idea, just trying to think it out from other angles, i like where this is going


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Re: Can psilocin be produced in a LC ala claviceps? [Re: KThunderland]
    #23844584 - 11/18/16 08:57 AM (7 years, 2 months ago)

No, I agree, it may not be viable. If the yield is not high enough or if it does not simplify the extraction enough then it wouldn't be worthwhile.

So far I've done the extraction 4 times. The first time I think I failed but I never sampled or tested the end product, it just didn't look like much of anything so I assumed it did not work. The 2nd time I think I had used 5 grams and I don't recall my yield but I would say it was anywhere from 50-100%. I think it was maybe 50mg of a brown syrup/oil. After that I did... 10g? I'm not sure, I think the yield was about the same tho and I got 100mg. The 4th time I did 100g of stumps, aborts, and dirt and I only got 100mg. I think a large part of the loss on the 4th try was wither because I did a poor initial extraction and did not use enough water/GAA or because I dried the organic with MgSO4 which is mildy acidic and held onto a lot of the alkaloids. I used MgSO4 on one of the previous extractions too but I did not use as much. I do KNOW some of this info is wrong, particularly the numbers, but that's as best as I can remember. 5mg of the extract is equal to maybe 0.7-1g of cubes I would say.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: krypto2000]
    #23845300 - 11/18/16 01:56 PM (7 years, 2 months ago)

i know one of the truffle farmers in the netherlands, he just uses it to finance his edible mushroom operation.  there's not near as much demand for pschedelic mushrooms as gourmet mushrooms.  I think he gets about 200kg of fresh truffles out of a cargo container and they go for about $200/kg wholesale so it's basically printing money...


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Re: Can psilocin be produced in a LC ala claviceps? [Re: drake89]
    #23846143 - 11/18/16 06:05 PM (7 years, 2 months ago)

Fun fact lol, a gourmet truffle farmer I spoke to in TN said he made about 250,000 in a yr


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Re: Can psilocin be produced in a LC ala claviceps? [Re: Raven44]
    #23847823 - 11/19/16 09:09 AM (7 years, 2 months ago)

Quote:

Raven44 said:
Fun fact lol, a gourmet truffle farmer I spoke to in TN said he made about 250,000 in a yr



He also has had no truffles the last 2 or 3 years due to climate change....more you know!


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Re: Can psilocin be produced in a LC ala claviceps? [Re: drake89]
    #23848280 - 11/19/16 11:30 AM (7 years, 2 months ago)

I have built some aeration vessels. Syringe filters do seem the best way to go whoever suggested that. I haven't made one that works in full yet, but I've made some first revisions which I later changed, but have not actually built yet. I'm still testing different glues as well, I'm not sure what is best.

Anyway here's the design I've come up with, it's pretty simple and versatile.  Take standard PP lids and drill holes large enough to wedge the narrow tip of the syringe filter through, it should hold snug if you did it right. Put two of these in and glue them in place. One is an inlet, one is an outlet. I've got some ~1inch air stones, some 3/16 inch silicone tubing which will fit on the airstone and on the narrow syringe port. It's not as tight as I would prefer on the syringe filter, but it's tight enough to do the job. Since it just slides on you can exchange it for the appropriate length so you can use these lids with any sized jar.

To power it I have a ~700GPH air pump with a 1/2" barb. The barb is designed to hook up a 1/2" tubing and connect it to a manifold with ~3/16" outlets, which would be idea to use, however the other side of the syringe filters are ~1/2" so I can't do this without adaptors. Instead I will build a manifold from barbed couplers and then connect 1/2" tubing to the inlet of the syringe filter. I've tested it and the variac is not necessary because it takes quite a bit of force to push through these filters and thus I'm thinking my ~700GPH pump is only powerful enough to power 1-3 vessels. I checked online and the pump runs for about ~25$ so it's not too bad if you had to buy it, a ~1700GPH pump is about 60$ which I would purchase instead if I were to start this new. So far the only potential issue that I might run into is I'm unsure what type of plastic makes up the barb on the air stone so it's possible it might decompose when PCing. If it does not work I could always use it without the airstone, but it would be nice to have as it will more efficiently diffuse and oxygenate the water better, while also probably being more gentle for the mycilium. Oh, and this pump is obnoxiously noisy so there is that drawback.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: drake89]
    #23848452 - 11/19/16 12:32 PM (7 years, 2 months ago)

Quote:

drake89 said:
Quote:

Raven44 said:
Fun fact lol, a gourmet truffle farmer I spoke to in TN said he made about 250,000 in a yr



He also has had no truffles the last 2 or 3 years due to climate change....more you know!




False

Last year he told me he made 250,000 from his truffles. Therefore whom I refer to has had truffles as of last year according to their word

Apparently u know someone else

That doesn't quite make sense. Climate change? I'm no truffle farmer but I do imagine the farming part means u create the climate as usual way farming.

Regardless, he has had truffles as of last yr. That's the only year we spoke in fact. Even if he was referring to the prior year which he wasn't that still wouldn't coincide not even close.

He was surely no liar, one of the smartest I've met


Edited by Raven44 (11/19/16 12:40 PM)


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Re: Can psilocin be produced in a LC ala claviceps? [Re: Raven44]
    #23848480 - 11/19/16 12:41 PM (7 years, 2 months ago)

You encourage the climate by moving water around, but if you live in a desert you live in a desert. Can't move water if there's no source.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: krypto2000]
    #23848564 - 11/19/16 01:04 PM (7 years, 2 months ago)

Yeah, I don't imagine this guy has any water issues

Sry to derail OP


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Re: Can psilocin be produced in a LC ala claviceps? [Re: krypto2000]
    #23848611 - 11/19/16 01:16 PM (7 years, 2 months ago)

Quote:

blackout said:
Growing was and is legal in many countries yet I have never heard of any using LC for production.





Quote:

krypto2000 said:
Those guys who make the atlantis truffles or w/e they are in amsterdam use spawn bags inoculated with LC. I saw a documentary or mini doc on it some years back.

Otherwise what you say is true, or likely so in the case of speculation.



I did not mean nobody used LC for growing mushrooms or sclerotia. I meant I never heard of anyone using LC directly for the production of psilocybin.

I was saying to have a search for threads of mine containing the word "keg", I give many ideas for liquid culture. In those thread I recommend using stainless steel or brass airstones which can be got in home brew supply stores. Most aquarium ones have plastic  or glue components which might not hold up to heating.

I think some filters will come with air flow limits. I have thin synthetic filters on my lids. I do not think they would keep out contams if air was forced through them at a high enough rate. There are other methods to prefilter air, like bubbling through peroxide.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: blackout]
    #23848658 - 11/19/16 01:27 PM (7 years, 2 months ago)

I didn't know they aerated beer, I'll check into those if these plastic ones do not hold up. I'm imagining they are some type of PP though based on the color and texture/hardness. Well I also don't *need* an airstone of course. I did just realize that in my test when I determined I could do 1-3 with my air pump I did not consider that it would also have to go back out the other syringe filter so that makes it only able to do 0.5-1.5, kinda lame. The syringe filter will definitely work though as this is more or less what it's designed for (pressurized filtration that is), but I might need a more powerful pump to do more than a quart of liquid. I was hoping to be able to run at least 5-10 pints at a time though as I figure I will have to screen a good 10-100 seperate cultures and combinations of media to get anywhere worthwhile. This is becoming a lot more daunting since I have to pioneer all of this stuff.

edit: Just tested it and I can definitely do 1 jar no problem, so maybe I'm still able to do 1-3. We'll see. I have one unit that is fully working though I don't trust the glue has dried enough to PC yet.

edit: And I just tested both existing vessels. One is not sealed properly, but running both seems to only reduce the flow rate by about 10-20% so I'm sure I can do at least 3 now. Maybe even up to 10, I should be good to go. :thumbup:


Edited by krypto2000 (11/19/16 01:46 PM)


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Re: Can psilocin be produced in a LC ala claviceps? [Re: krypto2000]
    #23855847 - 11/21/16 10:20 PM (7 years, 2 months ago)

Correct me if I am wrong but wasn't APE created via uv light mutation???

I wonder if the target traits were high be and potency like the ape have?

First it seems we need to know which psilocybe is most likely to produce high levels of psilocin in liquid fermentation


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Re: Can psilocin be produced in a LC ala claviceps? [Re: krypto2000]
    #23857669 - 11/22/16 03:11 PM (7 years, 2 months ago)

Quote:

Raven44 said:
Correct me if I am wrong but wasn't APE created via uv light mutation???




No, where did you hear this, the thread should be corrected if that info is in it, or maybe you just confused it. I think I have heard it rumoured about other albinos, possibly PF albino itself, which APE was developed from, it may not be true though, as there are reasons to lie about such things.

https://www.shroomery.org/forums/showflat.php/Number/5194890

Strain Origin: Hybrid of Penis Envy and PF Albino


Quote:

Raven44 said:
First it seems we need to know which psilocybe is most likely to produce high levels of psilocin in liquid fermentation



People should look at posts by Baby_Hitler -he is the most believable poster I have seen who posted about harvesting/dosing on myc, I think he recommended azures. Obviously people are already dosing on myc which has never grown shrooms, i.e. sclerotia. Some will swear on their mother's life that myc is inactive, especially if it has not fruited, maybe their mothers are already dead though! or more likely they have just never tried it.

I know RR frowned upon discussion of harvesting myc as in effect said the forum was about mushroom cultivation, not a "drug production forum", but discussion of sclerotia growing is not really frowned upon. I presume if an extremely easy way was discovered it would be more likely that spores would be made illegal though.


Quote:

krypto2000 said:
If you're growing it on grain you then have the biosmass + leftover grain to extract from, you might as well just grow mushrooms then.


the idea would be more like GLC, where you add water to colonised grains, wash it off, collect the myc strain it and let the jar recover. The grain jar can recover over and over again. It is far easier to identify if it is healthy looking growth on the grains than in a LC. If you are unsure you can shake the grains up and see if it recovers, while not 100% if it does not recover you can presume something is wrong.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: blackout]
    #23857778 - 11/22/16 03:36 PM (7 years, 2 months ago)

I'm not worried about contamination, if my culture on agar is clean then I know my LC is. LC is my main method of inoculation and I always have problems with them, but it's because I either get the filter wet or stick it with a dirty needle. Never had a LC grow with a contaminate in it, well no time recently anyway, it's always introduced later through using it. I won't have to even open these until it's harvest time.

Currently I'm a little set back on my tests. I made two LC lids and after PCing the aeration port no longer worked, I guess my syringe filter was not autoclavable : /.

edit: And to clarify on my LC us, I tend to make a quart or so at a time and will use it for months or even a year. They are always fine for at least a couple of months but then one day I'll see something in one of my jars or bags or on occasion if I haven't used it in awhile within the LC itself. A LC normally lasts me 3-12 months.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: krypto2000]
    #23857827 - 11/22/16 03:52 PM (7 years, 2 months ago)

Ty for the enlightenment blackout. Many good points and a good link.

I wish I could remember exactly where I heard that about APE sometimes my mind plays tricks...

Thank u for clarifying either way

Nexus comes to mind...


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Re: Can psilocin be produced in a LC ala claviceps? [Re: Raven44]
    #23858206 - 11/22/16 06:05 PM (7 years, 2 months ago)

Quote:

Raven44 said:
Ty for the enlightenment blackout. Many good points and a good link.

I wish I could remember exactly where I heard that about APE sometimes my mind plays tricks...




if interested

http://www.fanaticus.com/albino.htm
http://www.fanaticus.com/albino2.htm


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Re: Can psilocin be produced in a LC ala claviceps? [Re: blackout]
    #23868497 - 11/26/16 10:28 AM (7 years, 2 months ago)

So I got some new filters, ptfe this time, and the tiny air stones. I pressure cooked an airstone and a filter before assembling any this time to make sure they would not melt or decompose and they did not! I'm slightly concerned the glue on the air stone might leach into the broth and cause an issue, although it seems stable. I don't even have to glue in the filters either, I drilled my holes just right so they pop in there and are very secure and form what *seems* to me to be a good seal. On to some tests now. It might be awhile before I have any updates from here.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: krypto2000]
    #23869365 - 11/26/16 03:24 PM (7 years, 2 months ago)

Perhaps u could disassemble the air stone strip the glue and re assemble with a certain type of glue of ur choosing


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Re: Can psilocin be produced in a LC ala claviceps? [Re: Raven44]
    #23869512 - 11/26/16 04:10 PM (7 years, 2 months ago)

Well the glue did not seem to degrade at all so I think it's okay, I was mainly throwing it out there just as a possibility. It is targeted for aquariums too so one would hope they would use a non toxic glue.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: krypto2000]
    #23869551 - 11/26/16 04:23 PM (7 years, 2 months ago)

If the glue did give way you could just silicone it in place, most think of it purely as a sealant but its decent as adhesive.

I am going to be trying LC with potato starch as a thickener. Some say starch is not advisable, it is a far cheaper thickening agent than agar.

In other threads I mentioned the use of pumps, and internal submersed elements. Most will agree water can be sterilized by simple boiling. If you really feel the need to PC your other ingredients you can do this in a concentrated form in a PC. So you could PC a thick sugar syrup and then add to a larger homebrew style fermenter full of water, and boil this up again. You can buy tubs with heating elements preinstalled and holes and ports etc. My idea was to boil it up in the large container and have a pump running, you can get homebrew pumps intended for boiling sugary liquid. By running the pump you have boiling water passing through it and so sterilizing your lines at the same time.

The pump would simply by recirculating the liquid onto itself, it would draw liquid from the bottom and pump into the top of the container into open air, this would agitate the liquid, and aerate it at the same time. I would have a regular passive filter on the lid, as I said before I do not like the idea of forcing air through filters. At the top of the container where the liquid is coming back in you could have tubing with several holes in it, and plugged at the end, this would mean the liquid is coming out in several streams and so more aerated. I would worry about it clogging though, and the action of the liquid splashing onto the liquid below is probably more than enough agitation and stirring.


Quote:

krypto2000 said:
If you're growing it on grain you then have the biosmass + leftover grain to extract from, you might as well just grow mushrooms then.


A grain like popcorn is tolerant to repeated "washing and regrowth", you could colonise grain, introduce water and draw it off again which is just what GLC is, which I presume you are familiar with https://www.shroomery.org/forums/showflat.php/Number/6817701 . I do this in bulk with what I call syringe tubes, which are just silicone tubes with syringe needs on both ends. It allows you to transfer huge amounts of liquid with no need for a syringe.

I also have a jar I have yet to inoculate which is full of cut up silicone tubes, to which agar was added. This jar was heated so the agar was just liquid, and then shook about so the agar clings to the surface of all the silicone tubing inside. This is simiar to GLC but without the risk of grains, many think GLC is risky as the grains may have endospores.


Edited by blackout (11/26/16 04:39 PM)


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Re: Can psilocin be produced in a LC ala claviceps? [Re: blackout]
    #23998042 - 01/10/17 12:09 PM (7 years, 20 days ago)

Hey there,

SWIM is quite interested in the possibility to grow Cubensis solely in LC and check for Psilocybin production. SWIM started a small experiment of different Cubensis strains in liquid culture, which have been started from spores. Cultures have been grown in standard Malt Extract Medium with the addition of Peptone and Yeast extract (see BAM for reference). In order to oxygenate the mycelium, cultures have been placed on an orbital shaker and are grown in baffled flasks. They developed quite nice, check the pictures... Is this spherical growth a common observation?



So has this been done (What about OP?) and have the cultures been harvested and checked for alkaloid content (Quick literature research revealed nothing)? If so, do we know culture conditions and more importantly time point of harvest?

SWIM will be very happy to hear from you advanced mycologists!

Phen.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: phenethylamin]
    #23998069 - 01/10/17 12:17 PM (7 years, 20 days ago)

That growth looks pretty cool. I'm guessing that happens due to the shaking, the mycilium probably congregates in groups forming those spheres. I have only tried forced aeration and stirring though so I can't confirm. I have not had success in producing alkaloids so far and have since put the project on hold for a various reasons after the initiap failure. I plan to pick it up again but it may be awhile.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: krypto2000]
    #24011602 - 01/15/17 09:27 AM (7 years, 15 days ago)

Great idea.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: phenethylamin]
    #24014654 - 01/16/17 12:59 PM (7 years, 14 days ago)

Quote:

phenethylamin said:
SWIM started a small experiment of different Cubensis strains in liquid culture, which have been started from spores. Cultures have been grown in standard Malt Extract Medium with the addition of Peptone and Yeast extract (see BAM for reference).



was this spores direct to LC, if so you would want to test them. I have not seen growth which looks like yours before. I have seen small balls/clouds of myc though, which developed with a stirred LC, you can search for a user called blue helix for pics.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: blackout]
    #24014923 - 01/16/17 02:39 PM (7 years, 14 days ago)

Have you seen a LC grown on a rotary shaker before though? It makes sense why they would turn into balls, they keep turning and clumping together as they spin resulting in balls. Think of a wet ball of flour or something, as you spin it in a bowl the flour clumbs around it and forms a more or less perfect sphere (not really perfect of course, but within reason). I can't think of another example, but it's a common effect with small particles.


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Re: Can psilocin be produced in a LC ala claviceps? [Re: krypto2000]
    #24018483 - 01/17/17 08:34 PM (7 years, 13 days ago)

:takingnotes:


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Re: Can psilocin be produced in a LC ala claviceps? [Re: blackout]
    #24018979 - 01/18/17 01:23 AM (7 years, 12 days ago)
Log in to view attachment

yes this was a LC started from spores, which have been harvested from previous grows. This is definitely mycelia growing if you are in doubt of that, solid media has been successfully inoculated with the shown LC. Also there is no question about sterile work... :wink:

Different Cubensis strains have been observed and they all show this growth pattern, some tend to form smaller, others bigger spheres. Swim told me that the extraction of mycelia and liquid media is in process, so we have to wait for the analysis of that. In the meantime I did a more in depth literature research and of course Cubensis-LCs have been performed and results been published - see attached pdf/link and references mentioned there. Special interest should be payed to tryptamine supplemented cultures.

Source 1

Phen.


Edited by phenethylamin (01/18/17 01:24 AM)


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Re: Can psilocin be produced in a LC ala claviceps? [Re: phenethylamin]
    #24020018 - 01/18/17 01:38 PM (7 years, 12 days ago)

Word, thanks for that paper. :thumbsup:


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