ergot and agar

[bujinkan]

ok heres some interesting stuff.

GROWTH SPORULATION, AND ALKALOID PRODUCTION OF AN ISOLATE OF CLAVICEPS PURPUREA IN CULTURE:
L.Malik and D.Ganguly (regional res lab ., Jammu, India)

Growth and sporulation of the fungus were studied on plated containing different growth media.
 Both were poor on PDA and McCreas medium. On oatmeal agar + thiamine growth was totally submerged 
 and surface sporulation profuse. Kirchoff's medium + thiamine gave good growth and moderate sporulation.
 Presence of alkaloid could only be detected in in oatmeal agar + thiamine exposed to oxygen; a trace was produced in
 the absence of O. The amt. of alkaloid increased with the amount of added thiamine.

[bujinkan]

something else:

 DEEFINED MEDIUM FOR THE PRODUCTION OF LYSERGIC ACID ALKALOIDS BY CLAVICEPS PASPALI
Mizrahi, A.; Miller, George
(Israel Inst. Biol.)
The production medium, designated PM-2S, contained D-mannitol, 
50.0 succininc acid, 40.0, KH2PO4 2.0, and MgSO4.
7H2O 0.3 g/l.
 Usually, water was used and the PH was adjusted to 5.2-5.4 with NH4OH before sterilization. 
Defined media were prepared in deionized water. With certain additives,
 alkaloid production was as high as 505 µg/ml. 
The advantage of the medium with defined inorg salts is 
that the production of alkaloids is independant of variations in the compn. of water.

[bujinkan]

this one is interesting also

LYSERGIC ACID AND ITS DERIVATIVES BY FERMENTATION.
Rutschmann, Juerg; Kobel, Hans
The yeild of lysergic acid and its derivs. is enhanced by using Claviceps Paspali NRRL 3027 mutants as the 
fermenting organism. Such mutants are obtained by subjecting the microorganism to
 x-rays and or UV radiations and or ethyleneimine treatment. 
Thus, an active mutant was obtained by irradiating with a Hanau lamp 
the conidia (on malt agar surface) of the strain NRRL 3027 of C. paspali until .5-1% of
 the condia survived.
Cultures of the latter were suspended for 2 hrs. at 24deg. in 0.1% ethyleneimine soln. 
The spores were sepd. on a membrane filter,
 washed with sterile H2O, and seeded on agar plates. The ethyleneimine 
treatment was repeated 5-10 times. 
When cultivated on a medium containing 100g sorbitol, 36g succinic acid, 2g KH2PO4, o.3 g MgSO4, 1 mg of FeSO4, 
10 mg. ZnSO4 7-H2O (PH adjusted to 5.4 with NH4OH), the yeild was 2270 mg./1. of ergoline 
compounds of which 85% were amides of lysergic and isolysergic acids.

[bujinkan]

using highwire, found following on claviceps.

http://aem.asm.org/cgi/content/full/66/12/5419?maxtoshow=&HITS=&hits=&RESULTFORMAT=&fulltext=claviceps&searchid=1019487396775_6585&stored_search=&FIRSTINDEX=0&resourcetype=1

http://www.jbc.org/cgi/content/full/271/44/27524

please insert coin

[bujinkan]

http://www.jbc.org/cgi/content/full/271/44/27524?maxtoshow=&HITS=&hits=&RESULTFORMAT=&fulltext=ergotamine+synthesis&searchid=1019489824502_7398&stored_search=&FIRSTINDEX=0&resourcetype=1

only skimmed this one, might be useful. the sources look interesting, especially the ones referring to the mutant strains of claviceps utilized to produce optimal amounts of alkaloids.
please insert coin

[mottaman420]

Cultivation isn't hard its the finding of the initial mycelium that proves to be difficult.
The easiest to aquire medium i have seen is the following:
20% Cane Sugar
3% Peptone
Tap Water
PH adjusted to 6.2 with aquueous ammonia

The stated yields were 1.8 grams of ergotamine from 1 liter of substrate following an 8-10 day incubation at 24C.
Does anyone know where you can order claviceps paspali spores from??? Is it legal to buy the m like it is to buy cubensis spores or do they contain the desired alkaloids??
Blue Horses + Red Donkeys = Purple Mules

[bujinkan]

J** used to sell them, i think they can be acquired online...but id probably go looking in the surrounding farm areas first.
please insert coin

[Scooby_Doo]

A $10 bag of organic wheat sometimes has 2-3 little black grains. They are the same size as wheat just covered in a black looking fungi. Before having a little read I actually used to throw these little fuckers in the bin! They are probably something else but maybe just maybe.  

Apparently the specific gravity of the ergot covered wheat becomes less, this helps the farmers out since after transport they rise to the top and can be easily removed so that the wheat can pass regulations on the amount of ergot allowed to be present.

[GuySmiley]

   I was told in my other thread before it got locked that you could buy the egrot alkaloids from a pharmacutical manufacturer.....not sure if im getting that right.....you can legaly buy LSD precursors from a legitimate business? Also I asked how does one find ergot to cultivate, and dont have an answer, its certianly not in this thread. I read rhodium and looked at the last 15 pages of the tryptamine chem forum, so back of my butt about not looking enough.  
Cause its so fun!

[Rhodium]

The precursors cannot be bought legitimately, but they can be diverted from pharmaceutical businesses if you have the contacts.

[bujinkan]

guy smile
well, to discover the natural habitats of various claviceps species, look in a friggen encyclopedia. find out what ergot grows on, then find out if there are any fields of that stuff at a theater near you. get a little saavy...youll need plenty if you want to make our favorite chemical.

as far as buying or diverting goes...good luck. its doubtful that theres enough legit orders for yours to blend in with any sort of security...in the US anyways.

so there. we have no other options but to grow, and since everybody knows where to find samples, we are brought to the contents of this thread which begin to detail the process youll need to undertake.
please insert coin

[mellow]

IudexK2

Exactly how many thousands of pre-prepared agar plates have you got - How did you manage to obtain plates with claviceps media?

Seriously though: Everyone who grows this stuff grows it in liquid culture.

Agar plates are only used to produce pure cultures and for experiments (mutagenisis?). I used to like to get a sterile syringe with a small amount of saline in it and to gently scrape the surface of the agar in the vicinity of the mycelia. A small amount liquid is squirted on the plate and the shreaded mycelia drawn into the syringe. This is then used to innoculate a 250mL flask (several flasks) and these provide the starter cultures which can then be used to innoculate you main fermenters.

PS 1 - I repeat. No one grows claviceps on a large scale on agar. There's no reason to do that - so give up trying to get help - it's not done - unless you want to be the first, in which case, post your results in a few centuries time.

PS 2 - No I never did grow claviceps but the principle is the same for other fermentations.

[hest]

But you have to start the culture at an agar plate (and by the way It's a fast one, at least 10 times faster than psilocybe cubensis]

[carcrash]

I have heard of cubenses being grown in rice milk containers as sterile substrate for mycelium growth. Start with agar to get a pure culture then try a live tissue transfer to the rice milk container.

Grasping at straws on this one, but couldnt hurt to try. Atleast with the mushroom mycelium this is supposed to work.

I also wonder about doing a live culture transfer from agar to sterilized rye grain in myco bags. Pre soak the grain for 24 hours then pressure cook for an hour or more at 15 psi. I think the myco bags would be much better than jars here because you can thoroughly break up the substrate. Might need to use a bigger filter patch for air exchange.

Maybe in 10 years I will have enough knowledge to dream of lsd.

They are not cubes but they are fungis and I havent seen anything on anyone trying these ideas.

Ideal temperature and humidity parameters for maximum growth would help. With the ideal temperature parameter I would use an incubator made from 2 matching rubbermaid tubs 1 inside the other, the lower one full of water with a fish tank heater.

You cannot suceed if you don't try.  Learn from your failures.

[DrJackson]

SWIM long ago ran into a friend of friend of friend's dog who gave him the gift of a six pack. In this six pack was not killian's irish red, an amber lager which swim does enjoy, but a rather large quantity of claviceps paspali. Swim loves shroom gardening, and figured it was worth making the leap into chem. It grows well on potato water and sugar, but doesn't produce alot of amides. swim recently got bored and threw about half into a submerged culture, and will see what those yeilds shape up to be at his earliest convenience.

[sean1234]

http://www.ibiblio.org/herbmed/eclectic/bpc1911/claviceps.html

interesting, basic stuff on ergoline extractions from plant matter.

http://www.shaman-australis.com/~benjamin-thomas/Indole_in_Boletus_in_PNG.htm

this article has interesting ideas as well:

It is possible, as Ott (2000 [1997-1998]) has suggested, that the enzymes in some strains of Claviceps fungi are capable of converting lysergic acid hydroxyethylamide to the diethylamide when fed to submerged saprophytic cultures (Arcamone et al. 1961)

[MeNH2]

Try to use a microscope and ID it to make sure u are growing the right culter, and or othere tests .Ergot lookes differnt growing on grass and rye than it dose in culter. Make sure u are not making Butox . Bush might come after u .. hehe

[tomjuan]