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Ferather's Corner | Experiments, tests and data [Magic edition]


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#21 Ferather

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Posted 14 October 2017 - 12:43 PM

Ergosterol: [C-28]--[H-44]--[O]
Glycogen: [C-6]--[H-10]--[O-5]
 
 
Cellulose: [C-6]--[H-10]--[O-5] x z  (same as glycogen)
Starch: [C-6]--[H-10]--[O-5] x z  (same as glycogen)
Sugar: [C-z]--[H-2]z--[O-z]  (sugar varies)
 
z = Number of total units linked.
 
----
 
Baeocystin: [C-11]--[H-15]--[N-2]--[O-4]--[P]
Psilocybin: [C-12]--[H-17]--[N-2]--[O-4]--[P]
Psilocin: [C-12]--[H-16]--[N-2]--[O]
 
----
 
Serotonin: [C-10]--[H-12]--[N-2]--[O]
 
----
 
Potency is genetically capped, and also capped by nutrients.
Nitrogen and phosphorus, are key for active materials.
 
Yield could be lower, as carbon is consumed.


#22 Ferather

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Posted 14 October 2017 - 03:01 PM

Next test recipe for cubensis, dry spores:
 
pH: 7-7.5, roughly.
 
75g > Boiling hot water.
25g > Paper pellets.
2g > 240B gelatin.
1-2g > Sucrose.
 
The cubensis should activate on the sugar, it will also decay the proteins (from gelatin).
Hopefully there is enough other macro-micro nutrients present in the mixture.
 
There is ample carbon, nitrogen and water, essentially trace other.
If it germinates, I will be able to isolate cellulose growth.
 
----
 
I could use flour instead of sugar, for more nutrients.
However I would then need to fully sterilize.

Edited by Ferather, 14 October 2017 - 03:30 PM.


#23 Ferather

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Posted 15 October 2017 - 10:16 AM

Cubensis: 15/10/2017 | 16:00 | UK format.
 
The first image is pre-inoculation.
 
1.jpg 2.jpg


#24 mjroom

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Posted 24 October 2017 - 03:44 PM

I was reading in Keeper Trouts interview (https://trichoseriou...rview-with.html) regarding cactus and alkaloids the following statement that "A useful rule of thumb for nitrogen sources is that nitrates favor vegetative growth, while ammonium favors alkaloid production." I was wondering if this would hold true for mushrooms? any thoughts? Thanks. Regards mjroom.
 

 



#25 Ferather

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Posted 24 October 2017 - 04:02 PM

As far as I know, mycelium normally convert proteins and some vitamins into a soluble nitrogen format, which I think is Ammoniacal nitrogen (NH3-N)

If the case, mycelium will also convert other sources of nitrogen in the same way, so either way ammonical nitrogen, or ureic.

 

Nitrates: The main nitrate fertilizers are ammonium, sodium, potassium, and calcium salts.

This is different to a proteins or vitamins, as they are carbon based (linked).

 

1.jpg



#26 Ferather

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Posted 26 October 2017 - 04:03 PM

Cyan's on calcium bicarbonate (as carbon) + soluble micro-macro nutrients.
It took 6 days to get full activation, there is a lot of mycelium.
 
The recipe, is essentially just for spore activation.
 
IMG_20171026_214325.jpg IMG_20171026_214311.jpg
 
I will be testing the result.

Edited by Ferather, 26 October 2017 - 04:03 PM.

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#27 Ferather

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Posted 28 October 2017 - 03:14 PM

Data share:
 
Both samples where inoculated using a wood peg from the same culture.
The recipe's are void of starch and sugars, using only cellulose.
 
----
 
[Sample A]: 85g Water, 25g Paper Pellets, 1g MG, 0.12g YN, 0.5g Liquid Seaweed.
[Sample B]: 85g Water, 25g Paper Pellets, 1g MG, 0.12g YN.
 
Sample A:
 
Took 2-3 weeks to fully colonize, and a further 2-3 weeks to colonize 125g (dry), of the same recipe.
 
Sample B:
 
Took 4 months to fully colonize, and a further 2 months to colonize 125g (dry), same again.
 
----
 
Next test recipe:
 
85g Water, 25g Paper Pellets, 2g MG, 0.25g YN, 0.5g Liquid Seaweed.
 
Double strength, 18:1, it's essentially cellulose grains.
 
----
 

Seaweed: Beneficial bacteria, data here and here.
USDA: Seaweed, kelp, raw, 100g dry.

 


Edited by Ferather, 28 October 2017 - 05:03 PM.


#28 Ferather

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Posted 28 October 2017 - 03:24 PM

The entire recipe costs me 20p (UK), or £1.60 per kilo, dry. It does not require sterilization (no sugar or starch) and lowers the failure rate.

Please feel free to replace the brands and-or products I have used for alternatives with similar or the same ratios-results.

 

There are thousands of sources of macro-micro nutrients you need to add to wood, for example plant leaves.

Leaves can be very nitrogen rich, and mixing them with wood, is ideal for a complete substrate.

 

A good example is tea leaves, see here, which contain 3.1% nitrogen (rich).

They also contain ample macro-micro nutrients, and little sugars.

 

Leaves are acidic, so check and balance pH with CaCO3.



#29 Ferather

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Posted 01 November 2017 - 11:58 AM

I forgot I had these images, P.Cubensis var. Cambodian, brown rice spawn to WL-Tek.
And a fruit sample I took for testing, which looks rather strong to me.
 
No I did not eat them, it was hobby research!
 
PC 1.jpg PC 2.jpg
PC 3.jpg PC 4.jpg
PC 5.jpg PC 6.jpg

Edited by Ferather, 01 November 2017 - 04:27 PM.

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#30 Ferather

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Posted 01 November 2017 - 04:29 PM

I'm going to go ahead and say this, but please take no offense if you have tried it before.
 
DO NOT, ever, enrich products such as grain, with any type of solubles.
This includes sugar, you will increase risk of failure by 10 fold.
 
Save enriching for products that are otherwise weak.
Or use 50-60% spawn, and skip enriching.
 
----
 
Notes:
 
Cubensis utilizes cellulose just fine, you will need to calculate it's carbon content.
Additionally, it will utilize "soluble" macro-micro nutrients, as normal.
 
Other than no or little laccase, it's not much different.
It's primary to cellulose, and various proteins.
 
----
 
If a material cannot be made soluble (or small enough), it cannot be utilized.
Using utilizable soluble nutrients guarantees usage of the nutrients.
 
It's possible grains, wood, straw contain various insoluble's.
 
Spores love soluble nutrients + soluble carbon.
 
----
 
100% cellulose can be enriched with soluble macro-micro nutrients without risk.
If you add starch, sugar, CaCO3 (becomes soluble), be fully aseptic.
 
Cellulose is very complex and decay's very slowly.
Spawn provides the starch to grow well.
 
----
 
Cubensis primary enzyme list:
 
Proteins | Protease | Carbon and nitrogen source.
Cellulose, other | Cellulase | Carbon source.
Starch | Amylase | Carbon source.
 
Sugars, Carbon source.
 
----
 
Glycogen: [C-6]--[H-10]--[O-5]
 
 
Cellulose: [C-6]--[H-10]--[O-5] x z  (same as glycogen)
Starch: [C-6]--[H-10]--[O-5] x z  (same as glycogen)
Sugar: [C-z]--[H-2]z--[O-z]  (sugar varies)
 
z = Number of total units linked.
 
----
 
Paper pellets can hold up to 80% water content by dry weight.
However 75% was the better average, various brands.
 
100g dry x 3 = 300g water, 75%, roughly.


#31 Ferather

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Posted 03 November 2017 - 10:42 AM

Here I am testing another method of germinating dry spores from scratch, I'm using P. Cubensis var. Burma.

Recipe: 85g Water, 25g Paper Pellets, 2g MG, 0.25g YN. 1 x Cooked Wooden Peg, Total : £0.25.

 

The pellets contain soluble nutrients and complex cellulose, the peg contains the sugar.

 

IMG_20171103_143705.jpg IMG_20171103_143753.jpg

 

----

 

Making the peg:

 

I sterilized the peg in 50g water + 4g sucrose, to make a sugar peg (use tweezers).

Remove the peg, flame it to remove excess water, or use tissue and alcohol.

 

Using a dry spore print, scratch and collect some spores on the tip.

 

 

Making the sub:

 

Tip: Use boiling hot water, as it breaks up the paper pellets almost instantly.

 

Add the MG and YN to a jug, add the water, mix, add to the pellets.

Mix and mash for 5 minutes, microwave for 48 seconds.

 

Mix, and assemble, microwave for 48 seconds.

Add the lid, and allow cooling for 2 hours.

 

----

 

Infections should be limited to-around the sugar peg.
 
Hopefully growth will be vigorous from the start

Edited by Ferather, 03 November 2017 - 12:24 PM.


#32 Arathu

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Posted 04 November 2017 - 10:53 AM

Take a few of those to one of them wild stone circles you got there in the "old country" and hang with the spirits for a while...............hahahahaha................

 

As usual nice work man....... they do appear rather blue don't they?  :biggrin:

 

A


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#33 Ferather

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Posted 04 November 2017 - 02:20 PM

Nothing unusual, you wouldn't fruit either Oyster or Cyans (both lignicolous) on 100% grain, they taste bad, and don't put out.

Cubensis pretty much fails to detect and-or digest laccase requiring materials, but, it does detect-digest cellulose.

 

Gourmet oysters, are richer and more robust when grown from cellulose in optimal conditions.

 

Not all materials can be made soluble, using solubles almost guarantees usage.

For example, button mushrooms only detect certain proteins (nitrogen).



#34 Ferather

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Posted 04 November 2017 - 02:23 PM

Organically sourced soluble nutrients additives (free from starch and sugar, example fertilizer) do exist, I am not bias.



#35 Ferather

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Posted 04 November 2017 - 06:28 PM

Just to clarify the cellulose, glycogen and starch simplification:
 
 
Polysaccharide, source:
 
"When the repeating units in the polymer backbone are six-carbon monosaccharides, as is often the case, the general formula simplifies to (C6-H10-O5)n"
 
 
 
Cellulose, source:
 
"Cellulose is an organic compound with the formula (C6-H10-O5)n ... Cellulose was discovered in 1838 by the French chemist Anselme Payen, who isolated it from plant matter and determined its chemical formula"
 
 
Glycogen, source:
 
"The empirical formula for glycogen of (C6-H10-O5)n was established by Kekule in 1858"
 
 
Starch, source:
 
"Chemical formula (C6-H10-O5)n"
 
 
 
Simplifies to:
 
Monosaccharide, source:
 
"The general formula is (Cn-H2n-On)"
 
----
 
How to calculate dry weight:
 
Molecular Weight Calculator, select cellulose.

Edited by Ferather, 04 November 2017 - 06:29 PM.


#36 Ferather

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Posted 05 November 2017 - 03:50 PM

Some data I found in regards to microwaves, they are not perfect, however some positive info:
 
 
"Microwaves are as effective as conductive heating in killing B. subtilis spores, but the microwave E-field induces changes in the structural and/or molecular components of spores that differ from those attributable only to heat." -- Source
 
 
"The effects of this form of thermal excitation were studied in terms of morphological changes (light and electron microscopy) and also release of constituents (DNA and calcium ions) from the core of the spore.
Spores which are highly resistant to conventional heating such as autoclaving were fragmented by microwaves, albeit only at a greater intensity than was required for spores of mesophilic species." -- Source
 
 
"..bacterial endospores, which contain almost no water, are not destroyed by microwaves." -- Source (endospores must be hydrated, that doesn't mean it can't be HOT!)
 
----
 
General endospore data:
 
 
It takes 8-10 hours, for example, B. cereus to multiply by 1 million, that's a lot more endospores.
Also, it takes 8-10 hours to make a new endospore using 1 cell, how many will vary.
 
Quote: "Germination and growth generally occur between 10 °C and 50 °C".


#37 Ferather

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Posted 07 November 2017 - 11:07 AM

https://mycotopia.ne...-2#entry1342033

 

It's working @ 2.2% nitrogen, I have a tiny bit just breaching the surface, 4 days.
I will take images within the next 2 days for better visual quality.
 
Should also work for lignicolous mycelium, probs better.
I'm using WL-Tek max, grain ratios, 75% water.

Edited by Ferather, 07 November 2017 - 11:08 AM.


#38 Ferather

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Posted 07 November 2017 - 01:55 PM

I just about managed to get images of day 4. Young mycelium, on my "Sugar peg tek".
It's growing from the peg, no doubt for the sugar, no obvious contams.
 
Hopefully it will be better fed by day 7-8, no issues so far.
 
IMG_20171107_182942.jpg IMG_20171107_183301.jpg


#39 Ferather

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Posted 09 November 2017 - 06:30 AM

I transferred the cubensis peg, to verify cellulose genetics, it took only 2 days to regenerate and regrow.
The growth appears to prefer the enriched cellulose, speed is normal for this cellulose method.
 
Growth is more populated than oyster in the same environment, but less pushing out.
Refer to my cellulose agar alternative for start and end images of oyster.
 
As expected the sample and substrate(s) are clean of mold-other.
Can be used as a germination media, and pass to agar.
 
Slow to start, but it should eventually go pop! 
 
 
IMG_20171109_104606.jpg IMG_20171109_105117.jpg IMG_20171109_105309.jpg


#40 Ferather

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Posted 09 November 2017 - 07:42 AM

Here is some surface shots of 10% brown rice spawn to WL-Tek. The brown rice acts as the fast release nutrient source.
Spawn at the bottom, 15cm (6 inches) below, it took 2.5 weeks to fully colonize, [alternative method].
 
Ideally spawn should be clean, however a mild bacteria-yeast infection is "ok", but not ideal.
Yeast and bacteria can adapt to other carbon sources, even cellulose (plant fiber).
 
1.jpg 2.jpg
 
----
 
Cubensis notes:
 
Carbon sources: Cellulose, starch, monosaccharide's (sugars), other.
Nitrogen sources: Dung (ammoniacal, ureic), proteins, other.
Lacks: Laccase or detection of relevant materials.
 
Fruits from:
 
Dung with straw, plain straw and spawn, enriched soils.
Please note dung is a fertilizer, and is decayed.
 
Sub notes:
 
The preferred pH range appears to be 5.5-6.8.
Inhibited by plant defenses (extractives).

Edited by Ferather, 09 November 2017 - 07:58 AM.





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