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Offlinekrypto2000
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Are LAD producing strains of claviceps really that rare?
    #23806504 - 11/06/16 12:52 PM (7 years, 2 months ago)

Everything I've read seems to indicate they are, yet my experience seems contrary to that. I read, on one paper for instance, that various strains isolated from wild sclerotia as well as from culture libraries were grown out on agar and screened for LAD production and *none* produced detectable amounts of LAD without further selection and forced mutation. There's various reasons why this might be, but I won't get into it right now.

Just for shits shake I took 3 wild sclerotia I found, species unknown (paspali or purpurea I am assuming based on the location however), and plated them onto some agar in the same dish. I did not *see* any contamination, but I'm not sure. On the 2nd transfer of some I did notice some bacteria so it must be hidden in there somewhere. From these three sclerotia however I had at least ~5 different strains, probably 10-20 or more I would imagine as I have not yet noticed any sectoring and I'm on my third transfer from sclerotia. I have noticed various colored mycilium including white, white/beige, black/purple, green, and red/purple. The black and green might be the same strain/color, I'm not sure.

Anyway upon shining a blacklight on the bottom of the petris I noticed a blue florescence on the black/purple, green (I think), and white trays, I did not notice any on the white/beige or the red/purple trays. I took a wedge from both the white and the black/purple and extracted it with a small amount of MeOH which also fluoresced blue under black light. Upon reacting it with an ehrlich reagent the clear solution turned violet/pink. Perhaps this is a false indication, but I take it to be that these are LAD producing strains/cultures which means that at worst 20% of the strains I've grown produce a detectable amount of LAD which goes strongly against these other reports.

These were grown on standard PDA and nothing special was done to encourage LAD production. These tests were done on the 2nd transfer where the colors themselves were isolated, I'm not sure if I've isolated any single strains yet so I can't say whether that is the case or not. I will say that on the initial plate I had a large white growth area that did *not* fluoresce (and neither did any others) so I guess maybe the initial cultures did not produce anything as per the paper, but I assume they at least *isolated* the mycilium before drawing any conclusions. The ehrlich reagent is standard as well, I have not done any sort of calibration to know how sensitive it is, but I can say that fwiw you can definitely produce a more sensitive test than the one I used.


Edited by krypto2000 (11/06/16 01:01 PM)


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Offlinedrake89
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Re: Are LAD producing strains of claviceps really that rare? [Re: krypto2000]
    #23807158 - 11/06/16 03:40 PM (7 years, 2 months ago)

not sure if you're serious or a kid with a chemistry set.  BUT when you google "claviceps LAD" it comes up with a bunch of stuff about LSD.  So is that what you're talking about?  If so, please don't drink your broth it is likely toxic.


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Fiery Fungi (like us on faeboo)


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Offlinekrypto2000
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Re: Are LAD producing strains of claviceps really that rare? [Re: drake89]
    #23807180 - 11/06/16 03:47 PM (7 years, 2 months ago)

No I'm not drinking it. :macdre:

edit: Not sure what your question really is, but I'm not making LSD and definitely not drinking the stuff (though it would not kill you if you did, people seem to greatly exaggerate the dangers). The post is exactly what it seems, just tried growing this for fun/knowledge's sake, I have a test kit, so I tested them. This is the fungi that is used for LSD production among many other drugs, that's why I posed it in this forum, I figured some people might have experience or knowledge about it.


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Invisiblenooneman
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Re: Are LAD producing strains of claviceps really that rare? [Re: krypto2000]
    #23807197 - 11/06/16 03:53 PM (7 years, 2 months ago)

Most claviceps have some amount of ergot alkaloids, but like mushrooms it varies. If you take enough claviceps, you'll get ergot poisoning for sure, so that should tell you something right there. It's just a matter of the yields.

Sandoz had a high yield strain, specially grown just for that, but I imagine if you're in a bind any old claviceps will do. It'll just affect how much you'll end up with which for any regular chemist shouldn't be that big of a deal.


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Offlinekrypto2000
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Re: Are LAD producing strains of claviceps really that rare? [Re: nooneman]
    #23807209 - 11/06/16 03:56 PM (7 years, 2 months ago)

As I understood it most strains do not produce alkaloids at all, not just in small amounts. If the fungi makes it to the sclerotial stage there's a higher chance they will produce, but I don't know how likely or how much. This might well be true for psilocybes as well if you think about it, many strains are incapable of fruiting even so it would make sense they vary just as much. If they are unable to fruit they probably are unable to produce alkaloids as well. People who get gangrene also get it from eating a lot of sclerotia over a moderate period of time so it doesn't mean they all have alkaloids, just that a significant portion of them do.


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Invisiblenooneman
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Re: Are LAD producing strains of claviceps really that rare? [Re: krypto2000]
    #23807236 - 11/06/16 04:05 PM (7 years, 2 months ago)

I hadn't heard of them just not producing at all. It's definitely possible though if you think about multiple strains infecting the same grain. It can't be too rare though (20% sounds reasonable) because the risk of eating grain infected by ergot is quite real and will produce effects 100% of the time, but that could easily be explained by multiple strains infecting the same grain and 20% of those strains producing alkaloids.

Like you say, it's probably a lot like mushrooms, not all strains produce fruit, and the ones that do vary a lot, but still an MS inoculation of cubensis will produce some active fruits much as a "MS inoculation" of claviceps will produce some alkaloids.


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Offlinekrypto2000
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Re: Are LAD producing strains of claviceps really that rare? [Re: nooneman]
    #23807286 - 11/06/16 04:24 PM (7 years, 2 months ago)

Well not even that, but if you consider that ergotism take prolonged ingestion of the fungus and that it's ground into flour it could be well under that. Iirc in cases of ergotism it took weeks or maybe even months of eating the grain for it to occur. I recall reading during the worst times in europe back in the day when outbreaks would occur it was because they were eating flour that was up to ~25% ergot so they ingested quite a bit. They didn't have the sorting technology or ability to control its growth as they do today. I'm only referring to gangrene here, there are theories of histeria being caused by ergot injestion as well and that would be acute and not take time to build up, however most alkaloids are non psychoactive or they would eat below that threshold so usually gangrene is the first noticeable symptom. I'm recalling them from memory so I might be off on some things.


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Offlinekrypto2000
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Re: Are LAD producing strains of claviceps really that rare? [Re: krypto2000]
    #23847898 - 11/19/16 09:41 AM (7 years, 2 months ago)

Okay, so it might be that rare, I may not have found a single producer. I switched to a new kind of premade agar and noticed it fluoresced quite a bit under black light. I noticed that cultures growing on it will have flourescent bands, usually in the center of the culture, and one specific one had very bright bands on the outside. At first I thought this might be a sign of alkaloid production, but then I realized it might just be the compound within the agar being taken up so upon that realization I can't really trust my uv observations. I then decided to test an empty agar wedge with the ehrlich reagent just to be sure and what would you know it had the same reaction as the 'active' pieces. Oh well, this might be where this project ends.


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