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Invisiblepoboy
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Mycelial Culture Technique *DELETED*
    #3971708 - 03/26/05 09:47 AM (11 years, 8 months ago)

Post deleted by poboy

Reason for deletion: xxxx



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OfflineRogerRabbitM
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Re: Mycelial Culture Technique [Re: poboy]
    #3974925 - 03/27/05 02:46 AM (11 years, 8 months ago)

Mycelium has such little psilocybin, that tek is a waste in my opinion. You might get the equivilent of a 1 or 2 gram dose at best from a whole cake, when you could let it easily give you twice that if you just let it fruit invitro.


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Offlinecb9fl
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Re: Mycelial Culture Technique [Re: RogerRabbit]
    #3975646 - 03/27/05 12:05 PM (11 years, 8 months ago)

A BRF cake? I thought it would even be much lower than that.


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OfflineMindzpore
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Re: Mycelial Culture Technique [Re: cb9fl]
    #4016848 - 04/05/05 01:21 PM (11 years, 8 months ago)

maby if u use a sclerotia forming strain... but still uncertain.


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OfflineXTCollection
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Re: Mycelial Culture Technique [Re: Mindzpore]
    #4017960 - 04/05/05 05:06 PM (11 years, 8 months ago)

Mexi - A's are very potent when just ingesting the stones.

In fact, for strict mycelium extraction, AFOAF is switching to mexi-a's because azure and cubie mycelium can be hard to predict effectiveness. One night, a half pint sends you flying, another, takes 2.

Mexi-a's mycelium is very potent, as is it's sclerota...


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OfflineAbermelin
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Re: Mycelial Culture Technique [Re: XTCollection]
    #4018291 - 04/05/05 07:07 PM (11 years, 8 months ago)

Only mature mycellium contains psilocybin, meaning the cake would have to had flushed a few times, and from my experience, the psilo content in a cake can be pretty high... 7g-14g dried worth at least. Make a tea out of your old cakes. Growing mycellium in a liquid culture will yeild no psilo.


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Invisibletahoe
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Re: Mycelial Culture Technique [Re: Abermelin]
    #4018765 - 04/05/05 09:15 PM (11 years, 8 months ago)

if you do the tek, then use ps azure. It will have stronger mycelium. Dont go crazy witht his tek, its not as good as it seems. But 2 1/2 gallon jars full of mycelium wont be that much of a waste. The shit is hard to strain out, it clogs whatever you use istantly. You will end up with mycelium wafers. WE brought these to burning man 2 years ago and they went through security without a hitch.


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Offlinescatmanrav
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Re: Mycelial Culture Technique [Re: tahoe]
    #4019769 - 04/06/05 01:41 AM (11 years, 8 months ago)

>Only mature mycellium contains psilocybin, meaning the cake would have to had flushed a few times

Wrong and

>Growing mycellium in a liquid culture will yeild no psilo.

Wrong.

Although I dont think that trying to harvest psilo from a liquid culture would do you anywhere near as much good as a cake, far less mycelium.


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OfflineAbermelin
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Re: Mycelial Culture Technique [Re: scatmanrav]
    #4020049 - 04/06/05 03:07 AM (11 years, 8 months ago)

well, i could be wrong on whether the mycellium contains psilo inside a liquid culture, but i make that assumption based on the fact that:

a tea made with a fully colonized cake that has had no flushes will not give effects. trust me, ive tried.

Ive made tea out of a cake that had been through a few flushes and it had great effects.

It can be concluded then that what is meant by "mature" mycellium (If you read other literature on the subject, and not just one misinformed guide )is mycellium that has been through flushes. And since liquid culture has been through no flushes, it will contain only trace amounts.

But the tek you mention is correct in saying you will need to do gallons upon gallons of liquid culture to get a respective yeild that i would set at the value of one cakes worth of flushes/mycellium-psilo content.

Psilo producers tek = waste of time


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OfflineMindzpore
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Re: Mycelial Culture Technique [Re: Abermelin]
    #4020383 - 04/06/05 06:23 AM (11 years, 8 months ago)

Quote:

Abermelin said:
a tea made with a fully colonized cake that has had no flushes will not give effects. trust me, ive tried.

Ive made tea out of a cake that had been through a few flushes and it had great effects.





hate to be a nitpick but I instantly distrust any measurement based on your experience of something that you've cooked (assuming that you had to heat the cake to make tea). the cooking time, temperature and time you waited before you drank it would all affect the psilo-content.


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Offlinehyphae
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Re: Mycelial Culture Technique [Re: Mindzpore]
    #4020528 - 04/06/05 08:00 AM (11 years, 8 months ago)

Cakes bruise blue (fact). Blue equals actives.


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InvisibleArsey
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Re: Mycelial Culture Technique [Re: poboy]
    #4021819 - 04/06/05 03:49 PM (11 years, 8 months ago)

This site is too big for it's britches and hard to navigate... At any rate...
You have a study floating around here that was Published in Lloydia from the authors Catalfomo & Tyler that a few of you have obviously not read.

The paper deals with saprophytic culture which is basically submerged or liquid culture.

A few key points...

First...
Psilocybe Cyanescens did NOT produce Psilocybine under saprophytic conditions... considering azurescens may be closely related to cyanescens I would be leary as to it's usefullness in this type of culture. Ps. cubensis provided the most favorable results in the study

Second...
The peak concentration of Psilocybine occurs in liquid culture at the seven day point and drops of drastically after that point. The larger the volume of the flask the faster the rate of loss appears to occur.

Third...
Using the appropriate ph, the correct amount glucose and nutrients an average weight of 100mg of dry mycelium can be harvested from 30ml of nutrient broth. An average psilocybine content of 1% can be achieved. so...
300ml nutrient broth equates to 1g of dry mycelium
1g of dry mycelium at a conc. of 1% psilocybine equates to 10mg of Psilocybine
Which you can extrapolate to roughly +30mg per litre or +100mg per gallon.

Now if you are familiar with the studies of Gartz and Argurell you'll soon realize that you may be able to feed the saprophytic culture tyrptamine and potentially achieve a psilocybine level of 3%... a whooping +300mg of psilocybine per gallon.

Of course, the one complication you will have to overcome is the design of your bioreactors. The aeration of your culture becomes critical. You could run several stirred fernbachs if your goal is small. Otherwise it may be in your best interest to build something like a five liter airlift reactor out of PVC...

A five liter airlift would reasonable be able to produce 150mg of psilocybine and a possible yeild 450mg if it was enriched with tryptamine.. all in just about a week.


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OfflineHotnuts
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Re: Mycelial Culture Technique [Re: Abermelin]
    #4023419 - 04/06/05 10:24 PM (11 years, 8 months ago)

Quote:

Abermelin said:
Only mature mycellium contains psilocybin, meaning the cake would have to had flushed a few times, and from my experience, the psilo content in a cake can be pretty high... 7g-14g dried worth at least. Make a tea out of your old cakes. Growing mycellium in a liquid culture will yeild no psilo.




Notta! Been there, done that several times. It doesn't work. Not even close!!!!!!!


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Offlinedebianlinux
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Re: Mycelial Culture Technique [Re: Arsey]
    #4026455 - 04/07/05 03:34 PM (11 years, 8 months ago)

Quote:

Arsey said:
The paper deals with saprophytic culture which is basically submerged or liquid culture.




I couldn't even bother to read a couple sentences past this.

Saprophytic means a type of fungi that ingests dead or dying wood or otherwise lignacious substrates. Apparently the strict definition of saprophytic would place all fungi in the same category. As it is, saprophytes are just a sector of the fungi kingdom. Psilocybe Cubensis is not saprophytic. Shiitake, OTOH, is. Then you have saprophytes who border or cross into parasitism like Oysters and Honey Mushrooms.


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InvisibleArsey
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Re: Mycelial Culture Technique [Re: debianlinux]
    #4055747 - 04/14/05 06:56 PM (11 years, 7 months ago)

Quote:

debianlinux said:
Quote:

Arsey said:
The paper deals with saprophytic culture which is basically submerged or liquid culture.




I couldn't even bother to read a couple sentences past this.

Saprophytic means a type of fungi that ingests dead or dying wood or otherwise lignacious substrates. Apparently the strict definition of saprophytic would place all fungi in the same category. As it is, saprophytes are just a sector of the fungi kingdom. Psilocybe Cubensis is not saprophytic. Shiitake, OTOH, is. Then you have saprophytes who border or cross into parasitism like Oysters and Honey Mushrooms.




I'm sorry but your definition is not entirely correct.

Technically saprophytes are organisms whose nutrition involves uptake of dissolved organic material from decaying plant or animal matter not strictly wood and or lignin as you claim.

To "culture" means... growing a microorganisms, tissue cells, or other living matter in a specially prepared nutrient medium.

So if you dissolved organic plant or animal material into a liquid medium and feed it to a microorganism and it grew you have a saprophytic culture going on.

Would you like to tackle the next sentence?... Or maybe you'd like to write the national library of medicine and have them remove these ridiculous refs to Ps cubensis in 'saprophytic culture' like this one...

Neal JM, Benedict RG, Brady LR.
Interrelationship of phosphate nutrition, nitrogen metabolism, and accumulation of key secondary metabolites in saprophytic cultures of Psilocybe cubensis, Psilocybe cyanescens, and Panaeolus campanulatus.
J Pharm Sci. 1968 Oct;57(10):1661-7.


Edited by Arsey (04/14/05 07:06 PM)


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Offlinedebianlinux
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Re: Mycelial Culture Technique [Re: debianlinux]
    #4055811 - 04/14/05 07:20 PM (11 years, 7 months ago)

Quote:

debianlinux said:
Apparently the strict definition of saprophytic would place all fungi in the same category.




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Offlinegoofy98
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Re: Mycelial Culture Technique [Re: Arsey]
    #4056568 - 04/14/05 11:24 PM (11 years, 7 months ago)

i love it when a guy knows his shit(specialty).  It gives ya motivation to learn more. :playboy:


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InvisibleArsey
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Re: Mycelial Culture Technique [Re: debianlinux]
    #4058354 - 04/15/05 12:37 PM (11 years, 7 months ago)

Quote:

debianlinux said:
Quote:

debianlinux said:
Apparently the strict definition of saprophytic would place all fungi in the same category.







I was never trying to press the issue of classifying Ps. cubensis as to it's degreee of being a true saprophyte by english definition.

Apparently you were overwhelmed by the opportunity to exploit the semantics in an attempt to discredit my post, regardless, it appears you have lost sight of the actual issue being disscused here.

My intention was to discuss the prospect of producing fungal secondary metabolites in submerged culture. There are a few papers published by reputable institutions that already detail many aspects of doing this.

I feel that in a sense the OMC is somewhat begining to lack enthusiasm to potentially expoilt the use of common biotechnology. Submerged culture techniques are currently being employed all over the world to produce a vast array of substances from amino acids to secondary metabolites in both natural and genetically modified organisms.

There are countless designs of bioreators many of which can be improvised with relative ease. Many such devices are well suited for what the topic of this thread deals with. In fact, I'm very exicited about the concept of using modified spawn bags or even household oven bags and was hoping to bounce some ideas off of people who have some experience with a certian organism.

Aeration is a very important detail, the volume to surface area and rate of agitation have a significant bearing on the outcome of the culture... I haven't really seen anyone put this into prospective in this thread and I wanted to touch on that point.

Likewise, the nutrient content of the media and the duration the organism is cultured have significant bearing aswell.

Now if anybody would like to discuss building a poor mans bioreator, discuss fungal metabolism or submerged culture techniques I'm game. However, my english isn't so hot, my spelling is even worse and my grammer is pathetic so if you plan to pick me apart based on those details I'll simply go away.... by the way one more reason to stop at sentence two, I often leave out a word or a letter here and there when I type.


Edited by Arsey (04/15/05 12:44 PM)


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