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Glycerol, which can be present in the extracellular environment or within the cytosol at high concentrations, is a recurring motif in the physiology of extremophilic microbes. It is fungal xerophiles such as Aspergillus penicillioides and Xeromyces bisporus which dominate league tables for ability to grow in high‐solute environments and/or at low water‐availability (Stevenson et al., 2015a); these achievements can, in part, be attributed to their ability to accumulate and retain extraordinary levels of glycerol for osmotic adjustment. Along with some other microbes, these fungi are both capable of accumulating glycerol and are commonly associated with environments where glycerol reaches molar concentrations, including saline and sugar‐rich habitats; various types of fermentation milieu; foods, feeds and other manufactured products and within experimental systems (Hallsworth and Magan, 1994b; 1995; Wang et al., 2001; Patiño‐Vera et al., 2005; Bardavid et al., 2008; Basso et al., 2008; Donkin, 2008; Williams and Hallsworth, 2009; Chin et al., 2010; de Lima Alves et al., 2015; Lievens et al., 2015; Leong et al., 2015; Santos et al., 2015; Stevenson et al., 2015a). For instance, cells of fungi and algae can contain 7–8 M glycerol (see below); high intracellular glycerol is a determinant for vigour (Hallsworth and Magan, 1994a, 1995; de Jong et al., 1997); and the insect haemolymph, in which entomopathogenic fungi proliferate, can also contain glycerol at molar concentrations (Sformo et al., 2010). Studies of bacteria and fungi are carried out on culture‐media in the range 4–8 M glycerol (e.g. Santos et al., 2015; Stevenson et al., 2015a); and glycerol can also accumulate as a product in industrial systems (Wang et al., 2001; Cray et al., 2015a). Whereas in vitro studies of microbial solute stress typically focus on individual stressors, single‐solute systems are unrepresentative of extreme habitats found in nature (e.g. Lievens et al., 2015; Stevenson et al., 2015a; Yakimov et al., 2015). A recent study of extreme halophilic bacteria and Archaea, previously thought to have a 0.755 water‐activity limit for growth and metabolism (Grant, 2004; Kminek et al., 2010; Rummel et al., 2014), revealed cell division at 0.635 water activity with a theoretical minimum of 0.611 water activity, for cultures in mixed‐solute substrates (Stevenson et al., 2015a). Almost 70 years ago, a study of fungal xerophiles established a water‐activity limit of 0.640 for germination of Eurotium echinulatum conidia (a value equivalent to 64.0% equilibrium relative humidity); though the germination process was severely inhibited: germ‐tube formation only occurred after a 2‐year incubation period (Snow, 1949). Snow (1949) also reported evidence of a low level of (aborted) germination below this value: ‘One or two conidia … produced germ tubes at [0.620 water activity, though] many of the germ tubes produced were misshapen and probably not viable’. Other studies have reported germination for spores of X. bisporus, A. penicilloides, Xerochrysium xerophilum and other species in the range 0.740–0.700 which failed to yield any subsequent development of mycelium (Gock et al., 2003). Pitt and Christian (1968) reported limits of 0.644 and 0.605 water activity for germination of X. bisporus ascospores and aleuriospores respectively (though neither the authors of the original study nor ourselves have been able to repeat the aleuriospore study; data not shown).

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-> Errors Are A Great Source Of Knowledge <-
-> It Is Not Important WHO Is Right But WHAT Is Right <-

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(Yes, it maybe doesn't fit to this that well, but good general idea nevertheless)

But for sure, back to actual topic...

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FYI about the next quote, by John, what I meant was not that I am overall unhappy about the potential of medicinal mushrooms but rather how I'm concerned about something which doesn't just happen with mushrooms but many supplemental products. It's that early scientific research can often be oversold to the public, and people may apply too much of the potential to themselves. So I should have explained it is partially about using mushrooms as prophylactic medicine.
And most that are sold as supplements seem to relate to different kinds of medicinal applications as the antibiotics and anti-rejection compounds of the world.

About Ganoderma species I guess I asked more or less the same question as I did here:
https://www.shroomery.org/forums/showflat.php/Number/25893875

And about Chaga (as well as Agarikon) I asked about the medicinal compounds in LC which I have seen articles about.

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Sorry for the tardy responses, I've been traveling. I am in Ukraine at the moment, just arrived yesterday. Seems like I have too many projects running at any given time and not enough time to do them all...

There actually are a lot of clinical trials with mushroom compounds. Often though, it is knowing where the compounds came from. For example, take a look at any clinical trial on Statin drugs - they are from mushrooms. And all the anti-rejection drugs - also from mushrooms. And the current class of immune modulator drugs - mostly they are from mushrooms. For info on the latest cutting edge mushroom science, the journal "International Journal of Medicinal Mushrooms" is the standard scientific journal. If you are not familiar with this journal, I think you will find it interesting. And it is true there are many in-vitro studies linked to mushrooms. Actually, when you look deeper into it, you will find it is that way in any field related to pharmaceutical science. The first generation of testing is in-vitro, and referenced to the original starting materials, like a trial on mushroom species X. Then when the efficacy is shown (or suggested) in the in-vitro studies, further research is conducted under either the chemical name of the compound being tested or in the name of some trademarked designator name. Like the protein P40 for weight loss. It is the main protein found in a South African succulent plant called Hoodia gordonii, but you won't find any of the clinical trials with any reference to Hoodia. It is not in the interest of the ones funding the clinical trials to say where the compounds come from. This is where Science and Financial interests intersect. Most people do not understand why the trails would not be listed in the name of the material being tested. But as a scientist business man, I understand this implicitly. I could never fund a trial where we are testing something that could not be patented and return the investment in the trials. It is expensive to run a trial, believe me. In some cases we have spent more than $5 million US dollars on a trial for mushrooms, in that case, it was for a combo formula called Immune Assist Critical Care Formula. This is the heteropolysaccharides derived from 6 species of mushrooms, which we ran a number of clinical trials on. For a company to invest that much money on a clinical trial, there has to be some chance of getting a return on investment at some point. To just tell your stockholders "we are going to spend this much money on a clinical trial that anyone can use and benefit from", that just will not work. If we had more government money going into research, then we could do pure science, but as long as the trials are funded by companies, there has to be some return on investment. And this holds back the science. I wish it wasn't so, but that is today's reality.

regarding your questions on Ganoderma: The genus Ganoderma is a species complex as far as I am concerned. Rather than many discrete species, the genus shares so much common DNA sequence to be more of a large species complex. I recently ran the DNA sequences for D2 segment of 28s and also ITS1 and ITS2 segments. This was on a species I am well familiar with, I have been growing it for 20+ years and there is no question about its identity. The species in question was Ganoderma tsugai. I used it as a test subject in a DNA related study I did with the University of Nevada - Reno simply because it was so well identified that there was no question as to the actual species name. Yet in the three segments tested, for the top 100 matches in the NCBI genebank, there was not one single match to G. tsugai. It certainly matched a lot of other Ganoderma's, but not one match to tsugai. So when you ask if there are similar properties between the various Ganoderma species, the answer is absolutely yes. And there are no known toxic species, either in the genus Ganoderma or any closely related Genera like Phellinus.

As to ganoderma chemistry: We have identified well over 200 different structures of beta-bound polysaccharides, many of them based around a beta 1-3 glucan main chain, but by no means all of them. Many were, in fact, most were Heteropolysaccharides, with both alpha- and beta- bound sugars in the same molecule, and containing all manner of both 5 and 6 carbon sugars. So to call them all beta-glucans is simply incorrect. And the polysaccharides were NOT the same in different samples of the same species! In other words, nature has given the Ganoderma genus a wide variation in the normal compounds found. The polysaccharides are different in the mycelium and in the fruit bodies. What does this mean medically? That the larger the range of polysaccharide (PS) structures present, the broader the immune modulation activity. We humans, tend to think simplistically - one molecule, one effect. However, nature is not that simple. We can not refine this to "one active compound". It is the broad range of PS that gives Ganoderma its well know broad range of activity. In Chinese medicine, it is often read that for this problem, you use White Ganoderma, and for that problem, you use Black Ganoderma. Or red, or purple, etc. So what I did in formulating our best Ganoderma formula, called Gano Ultra, I combined equal parts of five different Ganoderma's, including lucidum, applanatum, tsugai, resinacium and another Ganoderma sp. (gold Ganoderma from Namibia, unknown species name). This has the best and broadest activity of any Ganoderma product I know of in the world today. I am not saying that other company's Ganoderma products are not good, just that this 5-Gano complex Aloha Medicinals makes has a very good record of efficacy. More than a million bottles have been sold, as well as several hundred thousand kilograms sold as raw material to other companies, and never one time has there been any negative feedback or side effects noted.

Your question on growing Chaga is a bit different though. Chaga, Innonotus obliquus, is an interesting fungus. The part that is traditionally used is tree-fungus interaction. I do not even know what this growth is properly called. It is not a fruit body, it is not a sclerotium, it is only partly myceliated, it is really the tree's own defense response to the fungal infection. The most well know Chaga, and the one with the most medical testing, is the one grown on White Birch trees in the Siberian birch belt. The White Birch is the species Betula papyrifera. And this tree, when it reacts to the Innonotus infection, produces a compound known as Betulinic Acid. This is a triterpine.

Wikipedia Description: Betulinic acid is a naturally occurring pentacyclic triterpenoid which has antiretroviral, antimalarial, and anti-inflammatory properties, as well as a more recently discovered potential as an anticancer agent, by inhibition of topoisomerase.
Molar mass: 456.7 g/mol
Formula: C30H48O3
ChemSpider ID: 58496
ChEMBL Id: 269277
PubChem CID: 64971
Index of Refraction: 1.533 chemspider.com

This compound was found early on in the research with Chaga. Since this is a unique compound, it was thought that this was "the" active ingredient in Chaga. Again, simplistic thinking. It is "an" active compound, but the truth is Chaga makes some of the most complex cell wall PS compounds of any mushroom, certainly some odd heteropolysaccharides. How much do these compounds influence Chaga's known efficacy? A lot I think. Now when you cultivate Chaga, no matter what you grow it on. you will not get any Betulinic Acid since this is only produced by the living birch tree. But you will get all the unique cell wall PS's. In my opinion, I think these probably contribute far more to the medical efficacy of the Chaga than does the Betulinic acid, but without isolating the compounds and testing each separately, who knows? And in any case, all the compounds present all add to or in some way medicate the medical efficacy of the product. Look at any natural product: Opium poppies are a good example. Any textbook will tell you that morphine is "The Active Ingredient". Well, that is not quite correct. Since there are over 50 closely related alkaloids present, and each of them contributes in some way to the efficacy of the product. In fact, raw opium on a weight per weight basis has 50% greater analgesic power (pain relief) than pure morphine, and it has far less respiratory and circulatory depression (side effects). We look at a single molecular entity as if it appeared from nowhere. But in fact to bio-synthesize any natural compound, there are pre-curser steps leading up to that target compound, and post-cursor steps leading away. If you have done any HPLC work, you will know that with any natural compound you get a classic mountain range shape to the graph, whereas with synthetic compounds you get a much sharper peak. Usually, you can just glance at an HPLC signature and now immediately if it is a natural compound or a synthetic compound. The reason for this mountain range shape is the pre- and post- cursers that are bio-synthesized along with the target "active" compound we are looking at. Does this make sense? Sometimes it is easy for me to understand a concept but not always easy to describe it to someone else so they can understand.

Anyway, I got off track. Back to Chaga: Yes, it is easy to cultivate. not difficult at all in fact. But you will not get the betulinic acid. Whether that is important or not I do not know. I have grown Chaga for many years, primarily on a substrate of organic white sorghum grain of a variety known as White Kaffir Korn. It is a popular product with clients. I have never run any clinical trials comparing it to wild-collected Chaga grown on birch, so I can't tell you how it compares. It is consistent though, which is the main quality looked at for today's marketing of any medicinally active compounds.

I hope these explanations help. More later.

John

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As for this next one: I probably don't have to tell you that I do think there are some problems with his line of thinking. While I do think that the symbiotic relationship of (ophio)cordyceps with insect species may not be exactly what most consider it to be at this point, his counter theories do not make sense and I don't mind pointing out the flaws in a reply.

That said, the information about boosting Cordycepin does seem helpful.

He also said mycotoxin content is among the lowest in the K1 strain.

I have doubts about needing to be so concerned about the mycotoxins, especially unless you are not taking serious dosages of Cordyceps like daily.
I've seen articles about testing of overall toxicity on rats and other animals and quite different concerning health effects were found like kidney damage, I believe.
No idea if they could have missed other kinds of toxicity in such trials by having used a commercial low mycotoxin strain I guess by accident, since they did not mention it.

The heterozygosity thing was about saving compatible monozygote cultures of ascomycete species in order to not have to worry anymore about losing ability to fruit due to trying to cultivate myc that turned monozygous.
Or I should probably say lose the willingness to fruit because it is probably an evolved protection mechanism against loss of genetic diversity / inbreeding sort of problems.

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Regarding the homo vs hetero - zygosity - as far as I know, it also applies to morels. That is the theory I am using in cultivating morels. I am not sure how to attach photos to this reply. If you tell me how to do that I can send you photos of the lab cultivated morels I am growing now at my farm in Minneapolis.

Cordyceps appears not to be a parasite in insects, at least Ophiocordyceps sinensis is not a parasite in Thitarodes moths so far as we can tell. Once an idea takes hold, it gets accepted as "fact" and that idea can be very hard to shake. When a dead insect sprouts a mushroom, it is easy to assume the mushroom was the cause of the insect's death. But why does this necessarily have to be so? What if the insect died, and then a mushroom grew from the dead insect? When you ay the "infected" host is not harmed, how do we define Infected? In fact, what appears to be the red blood cells in the Thitarodes moth, when you put them on agar, grow into Cordyceps. This is from any Thitarodes moth, healthy or not. So if all Thitarodes moths have naturally occurring Ophiocordyceps spp. present, how does this relate to an infection?

The human intestinal tract contains around 400 species of naturally mutualistic bacteria. Are these to be considered an infection? Parasites? or Symbionts? Or is it difficult to classify these mutualistic organisms? In truth, we evolved together with these bacteria. They perform required function in digestion, in immune function, in chemical signaling, and it is difficult to keep a human healthy without these bacteria. It seems to be much the same with Cordyceps species in insects and arachnids.

Supplementation in Cordyceps cultivation to increase Cordycepin production: The Cordycepin, 2'deoxyadenosine, is a breakdown product of the free adenosine produced during the growth phase. If the culture is maintained past the logarithmic growth phase, as the cells start to lys, the Cordycepin concentration increases as the Adenosine concentration decreases. this is the main method to increase the Cordycepin concentration - keep the cultures growing longer. This has a greater effect than any supplementation that we have found. In healthy, fast-growing (young) cultures, the Cordycepin concentration is pretty low. As the culture ages, it goes higher and higher. The peak seems to be reached in solid substrate fermentation when the substrate becomes so depleted in nutrients that the culture starts to fruit under high CO2 conditions. Once the culture starts to fruit, the Cordycepin concentration does not rise any more.

Brown rice is not a particularly good growth media for Cordyceps. It is lacking some essential nutrient that causes the growth rate to drop off drastically after about 14 - 18 days of growth, whereas with white sorghum, it does not drop off until after 30-40 days. Also, brown rice tend to be very sticky and therefore does not handle well in a larger scale growth method. Either millet, or Sorghum, or a blend of the two is a much better substrate for growing all types of Cordyceps. Oats works ok. Rye also works OK as a growth medium but does not give the same chemical profile so is not used for commercial production of Cordyceps spp. I have heard that Wild Rice (four species of grasses forming the genus Zizania) gives a good growth and chemica; profile but I have limited expereince with this substrate.

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I don't know about those journals, they seemed prohibitively expensive to me.

In any case John says he wants to provide the proper references for his earlier claims. It appears yeast-like symbionts are not so exotic as we might think. I need to combine a few PMs here but he provided this phylogenetic chart:



Also he says most insects don't use hemocyanin but I am not sure what that means for what he proposed regarding Cordyceps and its anamorphs.

He says to read this also: http://mjee.modares.ac.ir/article-3-3099-en.pdf

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In this study, we examined fungal symbionts variation of 13 populations of the insect vector of lime witches' broom phytoplasma. Molecular
phylogenetic analysis revealed that the fungal symbionts form a clade within the ascomycetous genus Ophiocordyceps consisting of entomoparasitic fungi. These results suggested the possibility that the fungal symbiont was acquired by the common ancestor of the subfamily Deltocephalinae and derived from an entomoparasite of the genus Ophiocordyceps (Nishino et al., 2016). The molecular phylogeny clearly showed that the fungal symbionts of the leafhopper vector were allied to the fungal symbionts of planthoppers. This phylogenetic pattern favors the idea of same evolutionary
origins of the fungal symbionts from the genus Ophiocordyceps within the Hemiptera, and highlights the evolutionary connection between parasitism and mutualism in the insect-microbe symbiotic associations (Hongoh and Ishikawa, 2000). It should be noted that the fungal symbiont of aphids is phylogenetically very close to the fungal symbiont of leafhoppers, although their evolutionary origins must be independentsuggesting the possibility that particular groups of Ophiocordyceps fungi specialized for hemipteran insects might recurrently serve as evolutionary
sources of the fungal symbionts in the Hemiptera
(Nishino et al., 2016)

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He is travelling a lot, hopefully more of a substantial update soon. The connection between parasitism and mutualism seems quite interesting.
I don't know yet how these relationships developed and whether Ophiocordyceps anamorphs are still (obligate) endosymbionts to certain families of insects or merely used to be.