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bodhisatta said:
<img src='/forums/images/moved.gif'> This thread was moved from Advanced Mycology.

Reason:
Host defense is essentially a placebo. In independent research that product tested the very worst compared to every other brand of mushroom supplement.

Most people use a double extraction.

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The Mycologist said:

He moved it because this is not an advanced subject.

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light4x said:

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bodhisatta said:
<img src='/forums/images/moved.gif'> This thread was moved from Advanced Mycology.

Reason:
Host defense is essentially a placebo. In independent research that product tested the very worst compared to every other brand of mushroom supplement.

Most people use a double extraction.

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You literally have no clue what you are talking about. The links to the studies are all in a huge thread on reddit. You definitely should not have moved this. Extremely poor and biased judgement on your part.

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There's plenty of threads in advanced that should be moved that get missed. This one didn't

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Mr. Alien said:

There are no actual mushrooms in Paul's supplements, they're based on grain/rice/mycelium biomass and contain mostly starch.

Just in case you don't believe what I say about it. Take it from Paul Stamets:

Here is an excerpt from one of Stamets’ patents:

EXAMPLE 9 [00123] The medicinal mushroom mycelium is grown utilizing liquid culture techniques. Whereas growing on rice might have 30-40% conversion of rice to mycelium, liquid vat culture may have essentially complete conversion with >3x more mycelium per unit mass.

30-40% conversion of rice to mycelium. Meaning 60-70% rice.

Check this research that confirms that 100%.

High alpha-glucan = starch which is from grain. While they will tell you the mycelium consumes the majority of the grain, the actual test data tells an entirely different story.

That is the reason they do not share actual lab test reports of their products. The same with Paul Stamets's products. Lot's of talk, but zero facts.

Their products are also not extracted, meaning the bioavailability is unpredictable for 80% of people.

Summarising: you are paying top dollars for useless starch.

IF you want to buy REAL mushroom supplements for your dad, I've only found 2 companies that actually SHARES! lab results. Very low starch content and high beta-glucans. They tell you the % of Beta-glucans. This is the second company! they use only fruiting bodies and they tell you exactly how much starch content there is in their products. Although, fruiting bodies are not where the most medicinal content is but is certainly better than Paul's products with 80% starch content or more.

Here is a quote from Oriveda.

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This information can be found on the product page for each mushroom. In some online communities the general idea is circulating that fruiting body is always preferable over mycelium. This is wrong.

In brief, here's why (it's mainly just common sense !):
for some mushrooms the most important active ingredients are actually in the mycelium (Lion’s Mane (erinacines), Turkey Tail (PSP/PSK), Cordyceps CS-4 (no fruiting body extracts exist!))
there is a difference between mycelium-on-grains/rice (biomass) and pure mycelium. Biomass should always be avoided as it is contaminated with undigested substrate in the form of starch. This includes all US-cultivated products. We only use pure mycelium, free from starch, if the products properties request the use of mycelium
quality claims are only worth something if they are backed with actual test reports. This includes a claim like 'fruiting body extracts are better than mycelium-based extracts' or vice versa. Nice statement, but where is the proof ? Also see our 'I'm confused by all the information online' entry, above. Using ‘we use fruiting bodies only!!’ has the subtext ‘it’s better!’ but actually only reveals the ignorance of the vendor, and the actual specifications, if any (% of glucans, terpenes etc.) usually have no verifiable source or backup in the form of official documents. Only specifying active ingredients is acceptable as proof of superior quality.

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Going back to fruiting bodies vs pure Mycelium for medicinal content.

Research has found that pure mycelium has most of the medicinal content! Companies like Oriveda uses a method called deep layer cultivate mycelia which is basically what we call "liquid culture" but they grow pure mycelium in MASSIVE amounts and extracts the medicinal content from there. NOT GRAINS. Companies like Paul and Aloha medicinals just want to sell you starch, is a FACT. They only care about money, not curing your mom or dad, if they would care they would share LAB TEST RESULTS. 

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filthyknees said:
Clearly there are different levels of effectiveness associated with different proccessing. Anyone with more than extremely basic research and mushroom knowledge would understand the myceliated rice may be more like a microdose, very clearly a different product than whole mushroom and of course extracts are MUCH different and for different applications.

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iwh678 said:

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filthyknees said:
Clearly there are different levels of effectiveness associated with different proccessing. Anyone with more than extremely basic research and mushroom knowledge would understand the myceliated rice may be more like a microdose, very clearly a different product than whole mushroom and of course extracts are MUCH different and for different applications.

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Tomorrow I'm going to start a 1 gallon liquid culture to get idea of what kind of yields you can from this method. I'll keep this thread updated on the progress.

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Wayward Place said:
I would like to comment on the breakingnof the cell wall issue. Now i am a noob but i am not dumb. As i understand it the mycelial structure is like a continous straw. So, if the mycelium, with or without rice ir starch, were broken in the grinding process, the the ends of each fragment of the straw-lime structure would be open for stomach acids to digest. If this is true, then the breaking of the cell wall, unless it co tains a valuable chemical, would be a pointless issue because the acids in your digestive track would hollow out the mycelial structure from both ends.

If i am wrong, please tell me and explain why.

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iwh678 said:
I ended up innocultaing two half gallon jars each with with 1.3 L of 4% LME.

I'm still new to making liquid cultures so I wouldn't be too surprised if I run into issues with these.

I also may end up doing g a few more quart jar with different mixes. I'm thinking I'll try a 2% LME LC and probably a grain water LC. Does anybody have experience with LC mixes that work well with links mane?

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dub rules said:
I am also not sure whether it is necessary to lyse the cells, but the supplements that Stamets sells under the Host Defense name are freeze dried. That process includes freezing and thawing under vacuum which I am assuming would cause the cells to lyse.

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No, freeze drying avoids the formation of ice crystals of the water present. And those ice crystals are what are responsible for screwing up tissue with regular freezing. And yes the freeze-thaw cycles.

Fungal cell walls are not only composed of chitin but also of polysaccharides which can be dealt with in various ways. The question is how this destabilizes the matrix and how big a molecule can be absorbed out of the cells how easily.



If we eat dried mushies we trip, right? But if you eat bigger chunks and don't chew well, its absorbed over a lot more time and the trip is weaker or rather stretched out.
Pulverized mycelium should be pretty good but freeze-thawing before drying and powdering may help make compounds more available but could potentially also mess up their structures so this is the tradeoff we are presented with, apparently. It can also make compounds more liable to degradation (for example oxidation) break down faster as it is not as well protected anymore inside the cells.

I know i am not staff, but idk which forum would be better for the discussion. It is about preparation of medicinal mushrooms, the angle is not to talk about chemistry tho sometimes it is hard to avoid.

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Fungusam said:
Have you seen this research paper about different liquids to use for the culture?
Here

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This paper has been my main point of reference so far:




As we can see from the study's conclusion, oatmeal is the best readily available option for making LC with a high erinacene content.
But how would one go about making an oatmeal LC that is expressly for this purpose?

Also, I'm having trouble figuring out the following table:




Is it saying that the end product would have the same concentration of erinacenes regardless of what concentration the media was,
or are the numbers in the table referring to the overall end amount? In other words, which concentration would net the largest total amount of erinacenes?

I'm looking forward to working together with y'all to figure this out!

So many aspects of this hobby haven't been thoroughly explored, so there is so much we can do that is on the cutting edge!

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iwh678 said:
Sorry for the late update guys.

Unfortunately there still isn't much progress on the liquid cultures. I think 4% was too high because they are growing awfully slow. I may try another culture with 2% LME this week.

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Fungusam said:
Pictured liquid culture is still steadily growing. I am going to start 6 new jars this weekend, but havent decided which solutions to use yet. Does anyone have suggestions? I'd prefer to make something that will be clearer this time, but at the same time be something that the lions mane mycelium will love to eat.

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Really not too sure tbh. Having a little read of the forums, most LC teks use a slow-burn sugar substrate - i.e. LME, because you want the mycelium to transfer to grains nicely. They mention something about not "shocking" mycelium that has grown on LC in a sugar substrate by transferring it to a complex sugar (grain), which is harder to metablise. That seems to be a key reason for choosing LME over sugars that metabolise faster.

Your goal is to just grow LC, so previous advice might not work in this case? This is a bit of a new project so maybe you should experiment? It is difficult because your intention is to maximise erinacine synthesis based on your substrate but, I am assuming so may be wrong, that you don't actually have the facilities to measure this once you have grown the LC?

I have tried to look into the literature but I have had trouble finding studies that are about growing mycelium. Are you uni-affiliated and able to get behind paywalls? We should try and do a literature review to get more information on techniques n shit. I can search the databases and try and find studies looking into this.

I had a look at that paper you linked. I gotta say their methods section is wildly confusing. Maybe I am just a bit dyslexic but it doesn't look to me that they actually fully explain their mediums - always a good sign in a paper
Firstly, they have a pre-culture medium which is a malt-yeast preparation. Then they transfer onto their experimental mediums, which I don't think are plain molasses, oatmeal, etc. The table you linked above looks to me like they had 8 mediums which were a combination of all the ingredients listed at those g/L amounts, minus certain ingredients. I think they were testing which components of the medium affected erinacine C production. That is how they get the preparation without oatmeal performing worse than preparations with oatmeal, i.e. prep. 3 vs prep 1 (which is the control preparation with all ingredients) respectively. The key performing preparations I think are prep 2 and prep 4.

It is annoying that they also had this mystery component, edamin K. I googled that and it is some weird powder that Merck produces to increase yield. The product page mentions a nitrogen analysis, so I am thinking it is some kind of fertiliser... thing.

I mention edamin K because the line graphs in the paper are comparing performance of edamin K with and w/o oatmeal in preps 7 and 8. They don't include prep 8 in the bar graph, which is curious. Seems they are hiding that result for some reason.

Anyway, if you are interested in trying different kinds of preparations, maybe try something like preps 2 and 4 from the paper? I would be interested in performance of preps 2 and 4, as well as four additional preps looking at removing the sucrose and mannitol (KISS - the fewer ingredients the better). Six preps all up.

1. Prep 2
2. Prep 2 minus sucrose
3. Prep 2 minus mannitol
4. Prep 4
5. Prep 4 minus sucrose
6. Prep 4 minus mannitol


The bar graph is interesting because it suggests that there is more eranacine production with less sugar in the preparations. Removing molasses and glucose have the biggest effects, but what if you removed molasses and sucrose, or molasses and mannitol, or glucose and sucrose, or glucose and mannitol? Again you probs don't have the facilities to measure eranacine production after these but they are a good start for thinking about this question anyway.

Ammonium sulphate and calcium carbonate are easy to come by, they are commonly used gardening compounds.

Also I just want to say, this paper is looking at maximising a single erinacine (C), when the mycelium produces tons of compounds. I think it is generally a mistake to try and isolate compounds from organic materials and then try and say this single one out of hundreds is the most important. There is going to be an important synergistic effect of all of the compounds in this mycelium, not just one they think has the greatest effect on nerve growth factor (NGF).

Anywho, that is my two-cents. I am super interested in this topic so will be perusing the literature. Will share my findings if I end up having any. Good luck with your current LCs, keen to see how they go.

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skywhisp said:
Still watching.

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Fungusam said:
Pictured liquid culture is still steadily growing. I am going to start 6 new jars this weekend, but havent decided which solutions to use yet. Does anyone have suggestions? I'd prefer to make something that will be clearer this time, but at the same time be something that the lions mane mycelium will love to eat.

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Really not too sure tbh. Having a little read of the forums, most LC teks use a slow-burn sugar substrate - i.e. LME, because you want the mycelium to transfer to grains nicely. They mention something about not "shocking" mycelium that has grown on LC in a sugar substrate by transferring it to a complex sugar (grain), which is harder to metablise. That seems to be a key reason for choosing LME over sugars that metabolise faster.

Your goal is to just grow LC, so previous advice might not work in this case? This is a bit of a new project so maybe you should experiment? It is difficult because your intention is to maximise erinacine synthesis based on your substrate but, I am assuming so may be wrong, that you don't actually have the facilities to measure this once you have grown the LC?

I have tried to look into the literature but I have had trouble finding studies that are about growing mycelium. Are you uni-affiliated and able to get behind paywalls? We should try and do a literature review to get more information on techniques n shit. I can search the databases and try and find studies looking into this.

I had a look at that paper you linked. I gotta say their methods section is wildly confusing. Maybe I am just a bit dyslexic but it doesn't look to me that they actually fully explain their mediums - always a good sign in a paper
Firstly, they have a pre-culture medium which is a malt-yeast preparation. Then they transfer onto their experimental mediums, which I don't think are plain molasses, oatmeal, etc. The table you linked above looks to me like they had 8 mediums which were a combination of all the ingredients listed at those g/L amounts, minus certain ingredients. I think they were testing which components of the medium affected erinacine C production. That is how they get the preparation without oatmeal performing worse than preparations with oatmeal, i.e. prep. 3 vs prep 1 (which is the control preparation with all ingredients) respectively. The key performing preparations I think are prep 2 and prep 4.

It is annoying that they also had this mystery component, edamin K. I googled that and it is some weird powder that Merck produces to increase yield. The product page mentions a nitrogen analysis, so I am thinking it is some kind of fertiliser... thing.

I mention edamin K because the line graphs in the paper are comparing performance of edamin K with and w/o oatmeal in preps 7 and 8. They don't include prep 8 in the bar graph, which is curious. Seems they are hiding that result for some reason.

Anyway, if you are interested in trying different kinds of preparations, maybe try something like preps 2 and 4 from the paper? I would be interested in performance of preps 2 and 4, as well as four additional preps looking at removing the sucrose and mannitol (KISS - the fewer ingredients the better). Six preps all up.

1. Prep 2
2. Prep 2 minus sucrose
3. Prep 2 minus mannitol
4. Prep 4
5. Prep 4 minus sucrose
6. Prep 4 minus mannitol


The bar graph is interesting because it suggests that there is more eranacine production with less sugar in the preparations. Removing molasses and glucose have the biggest effects, but what if you removed molasses and sucrose, or molasses and mannitol, or glucose and sucrose, or glucose and mannitol? Again you probs don't have the facilities to measure eranacine production after these but they are a good start for thinking about this question anyway.

Ammonium sulphate and calcium carbonate are easy to come by, they are commonly used gardening compounds.

Also I just want to say, this paper is looking at maximising a single erinacine (C), when the mycelium produces tons of compounds. I think it is generally a mistake to try and isolate compounds from organic materials and then try and say this single one out of hundreds is the most important. There is going to be an important synergistic effect of all of the compounds in this mycelium, not just one they think has the greatest effect on nerve growth factor (NGF).

Anywho, that is my two-cents. I am super interested in this topic so will be perusing the literature. Will share my findings if I end up having any. Good luck with your current LCs, keen to see how they go.

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What a thorough and well-written reply! Thank-you for your input    I will do my best to address all the points you've brought up.

Yes, I agree that some of the current advice on LC's is not applicable since this is not being transferred to grains. We can see that another experimenter in this thread tried using LME as the sole ingredient in their media preparation, and it didn't work out very well.

You are correct in your assumption that I don't have the means to measure Erinacene content, and you make a good point about different compounds having a synergistic effect. The main goal is to produce mycelium with a healthy percentage of beneficial compounds; so I'm not married to the idea of maximizing the yield of this sole Erinacene. However, I do think the percentage of erinacene C in this study could be an indicator of how well the production of other medicinal compounds is faring.

I have access to journal databases, but have never done a "literature review" or anything of that nature. You're right though, it would definitely be beneficial to get more info about "techniques n' shit"   I just don't know how to go about such a thing.

I'm glad it's not just me that found the methods section to be wildly confusing. I showed it to a friend of mine who is a fairly advanced amateur mycologist, and he had hard time understanding it too! It was so difficult trying to interpret how they were making those experimental mediums..

I'm not interested in using Edamin K. I'll be sticking with more commonly available ingredients, because one of the things I'm hoping to accomplish here is to create a Tek for the community. Once we make enough progress with this project, the knowledge that we've learned should be shared with everyone!! An obscure ingredient could definitely be a barrier to our fellows. So yeah I think it's best to avoid that if we can.

I have to admit, I've read through this paper a handful of times already, but I still don't understand how to reproduce the preparations described therein. I have six perfect LC lids ready to go for wide mouth quart jars, but I'm waiting to make anything until I know I'm making solid choices that will advance this experiment! Can you, or anyone, help me figure out many grams per liter of the ingredients to use for these preparations? No matter how much I study the table and corresponding description, I can't comprehend it enough to translate it to what I need to do.


As I touched on before, it would be a helpful to future growers if we stick with easily obtainable ingredients as much as possible. Maybe do something like that, but in a few different concentrations? i.e. honey, cane sugar, maybe a little LME, all those things w/ or w/o gypsum? The oatmeal LC I have right now is pretty foggy, which makes it difficult to identify whether there is any contamination. So I'm a little hesitant to continue using that as an ingredient (unless there's a way to make it into a clear solution, which I think might be impossible unless it's extremely diluted).

Regardless of which ingredients we choose for this experiment, I believe this study could be a great reference to determine what the optimal concentrations of different sugars are. If only we can interpret it!

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Shroomhunter53 said:
Don't forget that if you improve neural adaptation with disregard to the hedonic level personally and collectively you will become a parasite to the world.

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Shroomhunter53 said:
But this mushroom shows great promise to Severe Brain Injury patients in physical therapy.

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Fungusam said:
Also, I had a bicycle day thought. Would it be possible to grow mycelium in big buckets, kind of like how kombucha brewers do? Could it be a similar process? Or am I way off?

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Forrester said:

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Fungusam said:
Also, I had a bicycle day thought. Would it be possible to grow mycelium in big buckets, kind of like how kombucha brewers do? Could it be a similar process? Or am I way off?

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It would be nice, but I think it would be very difficult if not impossible, unless the buckets were inoculated in sterile conditions and sealed while fermenting.

Kombucha works for growing in such a nutrient-rich environment (there's a lot of sugar in there) because of the high acidity level mostly, (that's why when you first start brewing you have to use so much vinegar to start it or it will mold) and also because the kombucha culture has it's own bacteria and yeast that outcompete any unwanted ones very quickly.  After your first batch, the culture will produce it's own acid and keep the ph low enough to keep bacteria and molds from taking hold.  Also I think the scoby helps by sealing off the top from any spores landing, that's one of the reasons I believe it makes one.

I think to make something like this work with a mushroom mycelium, you would have to first have a mycelium that would be able to survive in a very acidic environment.  You'd also have to keep adding an acid of some sort because the mycelium will not produce it's own acid. 

Just my thoughts though, I could be wrong and maybe it's been done?

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Thanks for your input I knew very little about kombucha, so I learned a lot from your post

I was just looking over some research today, and incredibly, it turns out that lion's mane mycelium produces its own organic acids during growth! Which makes the growth medium significantly more acidic over time.

I didn't see this post before I was looking through papers, so I wasn't paying attention specifically to that aspect though. I think it's not uncommon for it to reach a pH of at least 4, if I'm remembering correctly. So maybe it is possible!

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Fungusam said:
Checking out some more research, I took some screenshots so I can share what I'm learning with anyone else who may be interested. Sorry that the formatting isn't so great, I don't have a lot of experience writing forum posts. Some of the info I've come across is quite thought-provoking!




This one came from an article titled "Erinacine C: A novel approach to produce the secondary metabolite by submerged cultivation of Hericium erinaceus". This reveals that Edamin K is no longer in production, and that its function in this context is to serve as a complex nitrogen source. Maybe this information can help us find alternatives?




This excerpt suggests that the pH of the media should be taken into consideration




Finally, an understandable version of a recipe! This is for the ideal media for erinacene C production. 2.73 g/L sounds like an impressive yield, especially considering it's only one of the compounds that the mycelium is producing




Some info about glucose concentration. This excerpt, as well as the following one, are from a the paper titled "Effects of Ascorbic Acid and Uracil on Exo-polysaccharide Production with Hericium erinaceus in Liquid Culture".




Wow, another comprehensible recipe! Really on a roll now, this is great
However, this recipe was formulated to optimize polysaccharide production. For the record, I am not yet familiar with what the function of the polysaccharides are. Or whether their production is an indicator of the presence of other medicinal compounds. So that's something else I/we should probably look into before moving forward

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Reish said:

This is all great information, but at about 2.5 g/l are you really getting more medicinal properties more efficiently than just growing the fruit bodies and just taking more as prescribed?

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Fungusam said:

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Reish said:

This is all great information, but at about 2.5 g/l are you really getting more medicinal properties more efficiently than just growing the fruit bodies and just taking more as prescribed?

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Just to clarify: that's 2.5 g/L of a single medicinal substance that was analyzed in the sample, not 2.5 g/l of biomass.

That substance was Erinacene C, one of the more frequently studied medicinal diterpinoids found in lion's mane. This substance is only found in the mycelium. So in the case of that specific substance, it definitely wouldn't be more efficient to grow fruit bodies, since the fruiting bodies don't contain any erinacenes at all. Hericenones are the aromatic compounds typically found in the fruiting bodies.

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Djchrist said:
Amazing thread

I was wondering if an extraction from the millet mycelium would be possible? Or would it work better from the fruit body?

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Again, you generally don't extract from mycelium.  That's like extracting the medicinal componenets from honey.  Most just eat the honey

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Forrester said:

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Djchrist said:
Amazing thread

I was wondering if an extraction from the millet mycelium would be possible? Or would it work better from the fruit body?

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Again, you generally don't extract from mycelium.  That's like extracting the medicinal componenets from honey.  Most just eat the honey

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Fungusam said:
Can someone point me to that John Holliday study? All I can find is stuff on cordyceps. I know that for example with John Stamets "mycelium" which is grown on grain, the end product is 60%+ starch.

Yeah I'd definitely prefer to do this in liquid. I'd prefer an end-product that is pure mycelium and extractable; and I'm pretty sure that you cant properly extract stuff that has grains in it. But I could be wrong.

Maybe someone can speak with more experience here, but when I think of lions mane mycelium, I think of myc that doesnt eat through all the grain before it starts trying to pin and put off fruits. And sometimes even starts fruiting before fully colonized. That doesnt happen in liquid though. Any thoughts?

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Fungusam said:
I know that with mushrooms you really want to extract them since they're not super bioavailable. Is the same not true for mycelium?

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No, it really isn't.  Look at pretty much every product Stamets sells, or Aloha medicinal, or 95% of all other mushroom supplements out there.  Mycelium grown on grain, dried, powdered, encapsulated.

Ever just munched a bag of the active mushrooms?  Worked fine, didn't they? 

And those are dried, tough fruit bodies.  Mycelium is microscopic thin strands.  Your stomach acid does a fine job extracting pretty much everything you need from it.

*** I'm not trying to knock your idea of growing in liquid culture, I just want to make sure it's going to be worth all the added effort, because it is going to be a lot of extra effort.  And I just don't see what the gains are, really...

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Forrester said:

*** I'm not trying to knock your idea of growing in liquid culture, I just want to make sure it's going to be worth all the added effort, because it is going to be a lot of extra effort.  And I just don't see what the gains are, really...

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Forrester said:
Nah I don't think you were off that it would be a better product, pure mycelium would be better if it were possible, it's just growing in liquid requires so much more effort and cost since it's so easily contaminated that way, and also more dangerous because contaminants aren't seen.  So you could easily be consuming something that wasn't lion's mane mycelium, without knowing, unless you test every batch.

I do remember the grain mentioned was either millet or milo/sorghum, so those might be a good place to start.  I used millet when I did mine, and it worked well.  A little tricky to get the moisture right for PC'ing without destroying the grain and ending up with mush, but it can be done.

For drying, I just cut up the whole substrate into slices after the prescribed amount of time, and put them in a food dehydrator, the same kind people use for cubes around here.  Once dried, a coffee grinder works fine.  The powder can be stored in mason jars and added to shakes or encapsulated if you want to take the time.

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Sorry, didn't see this before I submitted my last post!

Yes, I would definitely want to test each batch before consuming. However, can't contaminants also grow along with mycelium on grain, being virtually indetectable?

Edit: This is the method I was planning on using for cultivation of grain-based supplements. The main questions I have right now are listed above about how to proceed with the LC, but I will be attempting both of these methods either way

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Djchrist said:
Well regarding a lc batch you can always test it on agar to see where you at. Any results on the lc you did with the lion's main

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