Is that Aspergillus or spergillus ? I`ve never heard of spergillus and cant find it in scroogle .

"I think its important to know what other organisms might contaminate a submerged culture of claviceps"

Things that can get into the culture that will grow on the same substrate .

"There is a high osmotic pressure which I believe very few common organisms can live in."

What is the preasure ? If Claviceps can live in it why would other fungi and bacteria have a problem ?

"whos atmosphere should represent a properly cleaned lab."

Why ? A dentists is an open public room where fungi , bacteria and virii get in with the air wich isnt filtered and with all the , us , dirty contaminated bastards that wander in and out with no sterilility precautions .

Any culture would be made in a sterile maybe over-preasure lab and / or at least in a sterile over-preasure glove box . The tanks that sandoz grow it in are massive stainless steel silo like tanks ......... or at least that is how they grew it shown in the fotos i`ve seen . Those tanks would need to be agresively sterilised . I did a lot of asking years ago about it and was told that trying to grow and get ergot alkaloids from a liquid culture is a waste of time for anyone who hasnt got a proper lab , exactly the right claviceps and who cant test the claviceps used before its used for content . I was also told that they have to keep chooseing new cultures to use . Plus that to grow it properly it has to be kept moveing and be airated . That would make it nearly imposible to do for anyone without a lab .

The easyest way to get claviceps is from a flour mill

Youve probably all read it but there is a nice book out there called "Die mutterkorn-alkaloide" by uncle Albert ( <---- Hofmann ) .

The whole idea of  liquid cultivateing claviceps as a source of ergot alkaloids is stupid as its hard and easyer to get them from  / in countrys like india and / or by growing plants that produce them in their seeds there as has been shown recently by the busts on big LSD labs in india

The strangeness factor ------> If thats fuckin shit and a load of bolloks please tell me and point me in the right direction to find the facts .

(first batch was compromised, second batch was successful, characterized by ESI-MS.)
lately what I've found (under the microscope, and experimentally) is two possible factors which may inhibit alk production...the presence of [rod-shaped] endospore bacteria, i.e. bacillus spp., and mutation of a producer  strain to a nonproducer strain (this is noticeable from multiple passages on PDA plates). The former is for obvious reasons: competition for substrate, if it is actively colonizing. The latter is probably more of an issue, and one which needs to be addressed further. A paper which describes this is attached
*gives credit to Vespiary colleagues...thanks, guys*

UV-induced mutation is a roll of the dice, and arsenate salts are not easy to come by. what are some other options to regenerate the genetics for alk production?
It seems this issue is the most challenging aspect of the project, but it is also more amenable than, say,
getting wild-type c.purpurea to produce alks in submerged culture.

claviceps is a rather aggressive fungus, especially the nonproducer strain; it colonizes much more quickly than p.cubensis, so it's pretty resistant to bacterial competition (aside from bacillus, which can also thrive in the high osmotic pressure of ferm broth). However, it is the slower-colonizing darker variety of claviceps that is desirable for alk production.  

Lone Stranger, cocci bacteria are the ones which are inhibited in a high-osmotic pressure solution, as the lack the rigid cell-wall of bacillus; hence, they do not readily colonize. to determine what the osmotic pressure is of a solution, some solution density calculations are conducted. It's a widely accepted fact in microbiology that certain strains of fungus can not only grow in said solutions, but also alter their morphology. This is discussed in the the book "The Fungi" by Carlisle et al.

It's easy to knock an experiment, a la drone342, since there are easier methods out there; but you
really don't know unless you attempt the experiment yourself.
all the easy stuff has been beaten to death, pure science involves tackling problems.

It will be Aspergillus.

The easiest way to get Claviceps spp. is in the wild, on grass.

Liquid cultivation isnn't stupid at all, yes its hard, yes you have to revitalise your cultures, yes it involves working with some all-around pretty vile mutagenic chemicals.

Besides, a chindian could grass Tsathoggua up, if it choose to do so,  there are reporting procedures in place with (pseudo)ephedrine  sales you know, official agreements between hong kong-US, india-US and more, even extending in some cases, to trades that the US are never a party to, for reporting of sales to certain places.

He just doesn't have the faith in mankind that some people must do........people suck, they are greedy, venial, twofaced and shallow for the most part, if some chindian gets offered the chance to double his rations to TWO bowls of rice a week in exchange for a list of the names of his customers...do you think he isn't going to leap up and say 'yessir uncle sam thank you sir, now may I lick your boots clean in the name of holy ganesh while I'm at it?'

Some of the species occuring in that sample strike me as perhaps atypical, and perhaps not typical constituents of what might be flowing around say, an inner city, the countryside, your backgarden shed...it will vary.

Coprinus, Ganoderma I feel safe in saying will not pose a threat. those two are both saprophytic species growing mostly on dead wood, other unidentified basidiospores and ascospores are likely to be a lesser threat and belong to macrofungi. Most endospore forming bacteria are anaerobes which form spores to protect against destruction in aerobic conditions or other hostile environments, and will not reproduce in the highly oxygenated environment of a Claviceps culture (such as Anthrax, and Clostridium species)

I believe mutagenesis is essential, what strain was the one used by you, aniracetam? in every single paper I have ever read, wild type fungi produced the merest scraps of alkaloid in culture, we are talking 10-15mg/l.

Thanks for noticeing me trying to be helpfull and saying it . I was expecting lough to come in and be obnoxious again . .

I must admit that my atempts to grow ergot werent with liquid cultures and were in the 70s but since then i have kept watching for usefull information . LSD chemists and drug experts i`ve met said basicly what i am saying .

I dont say its imposible but it would take a lot of work , money and the returns would be small . The sterilisation / sterile work and airation again not imposible but dificult . The fact that one would have to find new cultures and test and identify them is also a no no factor .

The size of a container , steriliseing that container , steriliseing the substrate , keeping it all sterile and the small amount of ergot alkaloids that one would get would not be worth the time , effort , cost and dificultys as far as i`m concerned . It would be easyer to buy the acid here or fly to india and buy it there or produce seeds and use them for the synthesis .

I cant remember where i read the reports about labs in india but i think it might have been on necro-nutz and somewhere else . Scroogle says this http://www.mid-day.com/news/2010/oct/141010-USA-LSD-NCB-Gujarat-party-drug.htm Its not the report that i read but its about the same labs .

"Keep your cultures on cornsteep malt agar and transfer it every week or so, from the darker producing colonies.  Maybe every once in a while start a fresh culture from a dehydrated/lypholized master culture."

Sandoz dont do it in that for good reason = because that substarte is not specific enough and very many contaminants grow on it . The substrate/s they use have chemicals added to help the culture grow and so that it can produce more alkaloids . Getting those chemicals is also another cost and risk factor . The dificulty of transfering cultures adds to the possible contamination risks . That can be seen from a lot of the mushroom growing problems many people have with bought cultures that have been grown from cultures that have been grown from cultures and so on instead of produceing new cultures . The fact that the original ergot has to be changed is also a dificulty . Sandoz grows it in specialy inoculated fields and then identifys good strains every time they need to . .

aniracetam . Thanks for the info but the osmotic preasure dificultys are only aplicable to some contaminants and we dont get to choose wich ones we could have problems with . I aint knocking anyones efforts . Halleluja that people are trying . BUT i`m a practical person who is thinking about methods that can be used that work and not that are theory and have , sorry , little chance of haveing any results that would be usefull to anyone but a geek who is prepared to invest a lot of time and money and get very little return .

"people suck, they are greedy, venial, twofaced and shallow for the most part,"

LOL....... YUP they are a bunch of cunts mostly . The dificultys of obtaining things and with people opening their mouths are international and at least in india guards at factorys are bribable . The only real source of ergot alkaloids i have seen was from chemical firms that sold them and replaced a few grams in a kilo/s size order with something else . From those orders they got the ocasional 10 grams ...... but havent had any contact to them for years  .

SO gentelmen . I shall watch the reported progress here with interest BUT i would advise you to stop and go another way .

I appreciate the advice, Lone Stranger
but, I have the means, skillset, and determination to pull it off.

like I mentioned, all the easy stuff's been done. this is much more fascinating

(sorry about the pic quality, iphone 3G camera sucks)

A word of caution when using an iPhone camera: get software that removes metadata. You don't want your GPS location tagged to any of your photos.

I would also like to take this chance to say that I am a huge fan of your work.

TLS-I don't know.

And really, if we were interested in just growing LSA containing seeds, or buying acid from india, do you think we would be looking here? no.

Guys . The work has to be done and i`m glad your doing it . I hope you have sucsess and i volunteer to test it . I just like to play devils advokate and to look at things and see if i can see holes in them and better ways so dont take it to serious and dont let it discourage you . I aint slagging you off and will folow your progress wuth interest .

So striking Coprinus, Ganoderma and all the bacteria due to the use of gentamicin sulfate, what other fungi on that list pose no threat?  Can anyone think of a type of fungus which could be present and may pose a threat to a decently established culture of claviceps ?

Just do it all under a laminar flow, clean everything well with lysol or H2O2 before hand, and be diligent about the work. It shouldn't be to difficult to avoid contamination all together; and of course add a bit of gentamicin sulfate.