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After many years of experimentation, success has come to growing Claviceps in submerged culture:
http://www.geocities.com/cpaspali/cpaspali.html (http://www.geocities.com/cpaspali/cpaspali.html)
Claviceps is a unique organism, and to grow it submerged culture requires great attention to many parameters, such as strain genetics; media composition; fermentation parameters such as O2 levels, pH, temperature, proper nutrients, etc. Any deviation from the required parameters will result in no or poor growth of mycelium, or no production of ergot alkaloids.
Pictures show a strain of C. pasapli that produces Lysergic acid alpha-hydroxyethylamide as the primary product.
Key to this strains growth, and other C. pasapli strains, is large amounts of O2 in the liquid media available to the fungus. Air is pumped through 0.2 um PTFE filters to ensure sterile supply of O2.
The formation of "synnemata" or sclerotia-like form of the fungus is also a key parameter. Only sclerotia-like mycleium produce alkaloids in useable amounts; i.e. it is entirely possible to obtain large production of vegetative forms of mycelium that produce little or no amounts of alkaloids.
On the up side, by using synthetic rather than organic based media, contamination by orther micro-organisms such as molds, yeasts, and bacteria is reduced to a minimum.
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On the up side, by using synthetic rather than organic based media, contamination by orther micro-organisms such as molds, yeasts, and bacteria is reduced to a minimum.
Tell me more about synthetic mediums. What is it and how can I get it.
Cool pictures by the way. Its a pleasure to watch them.
Thanks
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If you search the Patent Databases for ergot, claviceps, etc. most that pertain to growing Claviceps will list specific formulas for synthetic media. Since each type of ergot fungus (C. pasapali, C. purpurea) and each strain has different needs, there are many to choose from and experiment with.
I'm using the term synthetic media in this context to denote a media where the nutrients are supplied primarily by inorganic chemicals (ammonium & nitrate ions)and alditols (sugar alcohols like sorbitol, etc.) rather than complex organic "foods" like soy beans, peas, meat & vegetable extracts, etc. The organic type medias can and do promote and support growth of unwanted bacteria, molds, yeasts, etc.
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Either it is contaminated or it is not. And that doesn't depend on your media but on your technique. So simply give your fungus what he likes most and what is the most convenient for you :) .
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I'm using the term synthetic media in this context to denote a media where the nutrients are supplied primarily by inorganic chemicals (ammonium & nitrate ions)and alditols (sugar alcohols like sorbitol, etc.)
Ooh do you mean that. I thought it was something new and high-tech or something.
:) Did you have a high yield of alkaloïds?
;D What formula did you used to make the mediums?
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growing Claviceps or any fungi. I use various media preparations, the "core" one based on Mannitol or Sorbitol, Ammonium Succinate, Magnesium Sulfate, and Potassium Phosphate. ( See
https://www.thevespiary.org/rhodium/Rhodium/pdf/arcamone.submerged.claviceps.paspali.pdf (https://www.thevespiary.org/rhodium/Rhodium/pdf/arcamone.submerged.claviceps.paspali.pdf)
and Patents as example)
With a good starting strain and ideal conditions, one can expect 1 to 2 grams raw ergot alkaloids per liter of media.
With good extraction techniques, one can expect to get 75-90% yield from that.
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The geocities page seems to be down.
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I copied the site to pdf for offline reading (on the metro). I could send or maybe post the pdf for everyone to see (with reference) if Bubbleplate wants.
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You certainly can make a PDF for the Bees here.
My apologies to those who couldn't access the Geocities Website. Yahoo Web builder leaves MUCH to be desired!
Try this new link:
http://www.geocities.com/cpaspali/ (http://www.geocities.com/cpaspali/)
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The page has been backed up to
https://www.thevespiary.org/rhodium/Rhodium/chemistry/articles/claviceps.paspali.html (https://www.thevespiary.org/rhodium/Rhodium/chemistry/articles/claviceps.paspali.html)
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Let's say that someone had a known strain on agar and wanted to keep it as it is. It's not possible to keep the agar plate forever, and copies of the strain will change over timer (genetic transformation).
Isn’t it possible to freeze dry my agar plate?
Hope someone with biological experience knows this
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You could store spores or sclerotia for a long time if you manage to induce their formation. It might also be possible to store viable mycelia in liquid nitrogen pretty much like bacteria or cell lines. Otherwise you have to keep your plates in a cold place wrapped with Parafilm and subculturing from time to time.
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I dont know of the viability of claviceps spp. spores, but generally i find that mycellium is a lot more viable than spores. I wouldn't bother keeping spores unless they are more viable than the mycellium. Storing the mycellium in a sterile syringe with a little bit of growing media (as some may be familliar with psilocybe cultivation) is a good idea. Psilocybe mycellium inside a syringe has known to be viable after 1 -year- or more after being stored at room temperature in a drawer away from light. Storing it in the fridge seems to reduce the viability time, reducing it to 2 months or so, i'm not sure why. Hope this helps.
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Hey now,
What ever gave you the idea that clav. pasapli is as forgiving.Im salvating at the thought of freezing under nitrogen.It gives you alot more freedom :P
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Spores are no good because they often lack their parents specific traits. The best way to store the cells is to induce the formation of conidia and freeze dry them. Their formation is quite easy and according to patent s can be stored indefinately. And is the exact genetic replication instead of many genitic variations found in spores. I'll find the patent I saw it in an dpost the info... It was one wher ethey used conidia to innolulate larger vats for mass production.