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


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#41 jkdeth

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Posted 09 November 2017 - 10:02 AM

I'm curious about that ph range. I was always taught that ph should be above 7, although part of the reason was to create an unfavorable environment for Trichoderma.
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#42 Ferather

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Posted 09 November 2017 - 11:53 AM

I'm not 100% sure of its default, but I know coir and grain are 5.5-6.8, below 7.

Increasing the pH to say 7.5-8 will lower chances of germination.

 

I am working with spores, so I went with pH 6.5.



#43 jkdeth

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Posted 09 November 2017 - 12:09 PM

Ok, I see, good info, the 7 plus I'm thinking about was generally for substrate. Although I just done a bunch of grain steeped in lime water at about 7.5. Too soon for results though.

#44 Ferather

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Posted 09 November 2017 - 12:12 PM

I like pH 7.5-8.5 for "live" oyster substrates, from reading cubensis is no different. You don't want spores after you have live mycelium.


Edited by Ferather, 09 November 2017 - 12:13 PM.


#45 Ferather

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Posted 09 November 2017 - 12:33 PM

I had better mention the down side to lime, lime can become soluble, although only instantly when it reacts, and else it's slow to react (over time).

While the higher pH deters spore germination, it also adds a soluble carbon source, which acts like a very weak soluble sugar.

 

I been successful with germinating spores using calcium carbonate (soluble end product), so it's not foolproof.

However, it should be no cause for concern on 100% cellulose, some concern with wood.

 

The lime causes decay of acids, which in turn can produce more solubles.

9 times out of 10, you will be successful with using lime.


Edited by Ferather, 09 November 2017 - 12:36 PM.


#46 jkdeth

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Posted 09 November 2017 - 01:19 PM

I should mention I've used your lime water on a stubborn pinner, with good results.
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#47 Ferather

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Posted 09 November 2017 - 03:28 PM

Yes CaCO3 water, or wood ash water, will trigger pinning with almost all types. I'm not 100% sure what causes it, here are a few reasons:

Reaction to sudden pH changes, sudden change in carbon-to-nitrogen ratios (soluble carbon) and decay of materials via pH.

 

Wood ash its self adds elemental "oxides", mycelium love oxides and O2 in general, they use it to decay matter.

All of this adds up to, either singularly or in combination with water, "I need to fruit" or "I am happy".

 

Pleurotus nebrodensis, will fruit specifically from limestone (CaCO3) rocks, yes rocks.

 

CaCO3 (inorganic), is also used by animals, to make eggs, or shells.

 

Ash.jpg



#48 Ferather

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Posted 09 November 2017 - 04:02 PM

I'm going to start using sugar pegs as standard.

 

IMG_20171109_200409.jpg



#49 jkdeth

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Posted 09 November 2017 - 05:20 PM

Any particular wood ash? Hard wood versus softwood etc.

#50 Ferather

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Posted 09 November 2017 - 05:23 PM

I don't think it matters, there is variation but they all contain CaO which converts to CaCO3.



#51 Ferather

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Posted 15 November 2017 - 11:25 AM

Exports:
 
----
 
13/11/17 Uk format, I assembled ~14% sucrose and water, 4g to 25g water, into a syringe, made clean.
I used it to sugar drop my peg, cellulose as carbon is too complex for spores used alone.
 
I am using WL-Tek [Enrichment tek], which add's nutrients, but no carbon.
 
IMG_20171113_142322.jpg
 
----
 
Yesterday (14/11/17, UK format):
 
Finally got the response I was looking for, it seems the sugar peg is mostly good for germinating.
After sugar dropping my test sample with ~14% sucrose water, it has regenerated better.
 
Growth is about twice as lively than on the trace sugars in the peg.
Still no contamination, just weak cubensis on a peg.
 
IMG_20171114_190630.jpg IMG_20171114_190857.jpg
 
----
 
Today (15/11/17, UK format):
 
Cubensis var. Burma, dry spores onto WL-Tek:
 
The 14% sugar solution, is being spent rapidly as expected.
My cubensis has accelerated, overnight.
 
IMG_20171115_123906.jpg
 
Heat production on site too.
 
IMG_20171115_124045.jpg

Edited by Ferather, 15 November 2017 - 11:28 AM.


#52 Ferather

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Posted 15 November 2017 - 05:37 PM

Here is an easy comparison:
 
Substrate recipe:
 
> 85g Boiling Hot Water, 25g Paper Pellets, 2g MG, 0.25g YN.
> Sturdy microwave container, I'm using a micro PC.
 
----
 
[Sample A]: Tarragon oyster (lignicolous), small tissue and paper sample (no need to germinate), two weeks to accumulate enough resources, to be "weak".
[Sample B]: Cubensis, from dry spores, sugar (sucrose) assisted to provide soluble starter resources, and improve growth and speed.
 
Both samples where assembled onto the same substrate, at roughly 2.4% nitrogen, approximately 20:1 carbon-nitrogen.
The "soluble" additives also provide other needed essential macro-micro nutrients, but no carbon source.
 
Both samples where assembled in fully exposed conditions, with [Sample A] having a film lid.
The film lid has added holes, and no filters, it is covered with dust and debris.
 
Neither sample has produced any visible signs of third party decay.
 
 
Images: [Sample A]:
 
IMG_20171115_214135.jpg IMG_20171115_214327.jpg
 
Images: [Sample B]:
 
IMG_20171115_213421.jpg IMG_20171115_213523.jpg
 
----
 
The products being used are intended to be added to "carbon" rich materials.
Which is why "plant food" is not a food, but however an additive.
 
MG - Soil, adds no carbon to the end media.
YN - Beer making, adds trace carbon.


#53 Ferather

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Posted 16 November 2017 - 03:14 PM

Exports, 16/11/17 UK format:
 
----
 
Cubensis var. Burma, WL-Tek @ 2.4% nitrogen. Growth is visibly similar to rich grain, but slower.
This is due to cellulose, decay is improved via addition of a fast release glucose source.
 
The added macro-micro nutrients are near enough equal to grains ratios, 20:1.
 
50:50 dry-to-dry spawn to WL-Tek @ 2.4%, should be equal to all grain.
Except the cellulose, which will decay faster due to the starch.
 
Enriched cellulose, should not contaminate alone.
 
IMG_20171116_170505.jpg
 
----
 
Final notes:
 
Cubensis produces cellulase, however like all other mycelium which produce cellulase, it must be assisted, it's too complex (slow release).
Without huge experimentation, the two easiest ones are starch-sugars, in terms of the safest, trace starch as sugar is too "ready".
 
Sugar (sucrose, in this case), should be utilized my almost all organisms, whereas starch has a chance of partial decay.
There are 3 main types of starch enzymes, some will only utilize a small portion of starch, some decay all of it.
 
Cubensis utilizes the additives in the same way the intended organism does, onto carbon rich products.
However, do not, enrich grain-seed with any product, it's simply not needed and a bad idea *.
 
Enrichment, as seen commonly in gourmet growing, is for cold (low nutrient) products.
 
* Other than for grit, or to capture falling condensate (verm).
 
 
Note: Amylase works best at a pH of 6.7–7.0.
 
----
 
Also here are today's surface images.
 
IMG_20171116_190409.jpg IMG_20171116_191704.jpg


#54 Ferather

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Posted 17 November 2017 - 05:20 PM

Free guide (export from another forum):    Attached File  Ferather's Journal.zip   1.15MB   44 downloads

 

Thanks everyone for your posts, images and data, enjoy my consolidation!



#55 Ferather

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Posted 30 April 2019 - 06:57 PM

27/04/19
 
Azure wood sample to T-Gel plain (no additives), test:
 
IMG_20190427_003650.jpg IMG_20190427_003745.jpg
 
----
 
29/04/19
 
Azure sample to T-Gel, plain, no additives, 3 days:
 
The increased oxygen has made all the difference, the previous test took 7 days to respond, and then stalled after 11 days.
Currently I am using the same oxygen value I used in my Cubensis test, the results are more normal.
 
Un-adapted first transfer oyster mycelium takes around 3-4 days to respond.
 
IMG_20190429_214509.jpg IMG_20190429_214547.jpg
 
The black tint is oxidation.
 
----
 
30/04/19
 
My Azure sample to T-Gel, plain (no additives), has landed exactly within the 3-4 day window.
Growth is linear and populated, not rhizomorphic, it's also quite quick.
 
Reminder: There is no sugar or starch in T-Gel.
 
IMG_20190430_215723.jpg IMG_20190430_215924.jpg


#56 Ferather

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Posted 30 April 2019 - 07:10 PM

T-Gel Agar
 
This is not a one shot clean all method, however it does limit bacterial activity.
 
----
 
 
 
----
 
Recipe:
 
100g > Water + Cook Water  |  2g > High Quality Agar Powder  |  3.25g > Black Tea Bag  |  Optional: 1g > Malt Extract.
 
Black tea should be used, it has already undergone oxidization, and produces more solubles.
 
I am using Intralabs's 100% pure food grade agar.
 
----
 
Bacterial peg to T-Gel, the bacteria was slow, and mostly overrun. It required 3 transfers and validation.
 
IMG_20171207_232124.jpg     IMG_20171207_232251.jpg     IMG_20171207_232348.jpg     IMG_20171012_213838.jpg
 
I suggest using standard (more clear) agar recipe's for validation.
 
----
 
Cubensis Burma on T-Gel (no ME), 3rd transfer:
 
The temperature is ~8°C, and the Cubensis has produced a white-rot effect, due to enzymes-reactants (see here).
 
Decay and absorption goes beyond the physical mycelium, it's also producing heat.
 
IMG_20190308_154526.jpg     IMG_20190308_154617.jpg     IMG_20190308_154712.jpg
 
IMG_20190413_112757.jpg     IMG_20190413_113014.jpg     IMG_20190413_113143.jpg
 
The end pH is around 6.0, using hard water.
 
 
Here is a single wedge to brown rice.
 
IMG_20190205_133013.jpg     IMG_20190205_133120.jpg
 
IMG_20190208_202026.jpg     IMG_20190208_202038.jpg
 
----
 
Nutrients:
 
 
 
Proteins, varies:  [ Cx | Hx | Ox | Nx ] x. -- Trace, minimal.
Polyphenols, varies:  [ Cx | Hx | Ox ] x. -- Phenols.
Cellulose: [ C6 | H10 | O5 ] x. -- Trace, debris.
 
Theobromine:  C7 | H8 | N4 | O2. -- Methylxanthine.
Theophylline:  C7 | H8 | N4 | O2. -- Methylxanthine.
Caffeine:  C8 | H10 | N4 | O2. -- Methylxanthine.
 
Theaflavin:  C29 | H24 | O12. -- Polyphenol.
Tannins:  C76 | H52 | O46. -- Polyphenol.
Catechin:  C15 | H14 | O6. -- Phenol.
Flavonoids:  Cx | Hx | Ox. -- Phenol.
 
Carotene:  C40 | H56. -- Hydrocarbon.

Edited by Ferather, 30 April 2019 - 07:30 PM.

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#57 Ferather

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Posted 02 May 2019 - 08:59 AM

Azure sample to T-Gel, plain (no additives):
 
Visible oxidation (black ring), which is normal (tea turns black as it oxidizes).
 
IMG_20190502_144332.jpg IMG_20190502_145358.jpg IMG_20190502_145404.jpg

Edited by Ferather, 02 May 2019 - 09:01 AM.


#58 Ferather

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Posted 07 May 2019 - 06:37 AM

Azure sample to T-Gel:
 
Fresh growth has a different colour from the original growth on the wood, it's more blue-black-green.
In addition best growth is growing towards GE, where the front has the highest exchange.
 
IMG_20190503_151201.jpg IMG_20190503_151341.jpg IMG_20190503_151510.jpg
 
----
 
Extract from my journal:
 
Baeocystin:  [C11]--[H15]--[N2]--[O4]--[P]
Norbaeocystin: [C10]--[H13]--[N2]--[O4]--[P]
 
Psilocybin:  [C12]--[H17]--[N2]--[O4]--[P]
Psilocin:  [C12]--[H16]--[N2]--[O]
 
 
"Psilocybin is a member of the tryptophan-based compounds that originally functioned as antioxidants in earlier life forms before assuming more complex functions in multicellular organisms."
 
"Psilocin is relatively unstable in solution due to its phenolic hydroxy (-OH) group. In the presence of oxygen it readily forms bluish and dark black degradation products"
"Most species of psilocybin-containing mushrooms bruise blue when handled or damaged due to the oxidization of phenolic compounds"
 
----
 
Since phenols require oxidative agents, such as laccase or radicals, the mycelium likely produces more anti-oxidants.
Increasing the tolerance to oxidants, increases the rate, and ability to decay with oxygen.
 
 
----
 
Various lignicolous mycelium such as oyster's decay phenols using laccase and peroxidases, such as lignin peroxidase.
Some methods may require oxygen to function properly, in order to "oxidize" phenols-other (decay them).
 
Here is Tarragon oyster on phenol based T-Gel Agar (plain, no further additives).
About 3.6mm of the surface has turned clear, not much remains.
 
IMG_20180125_124225.jpg IMG_20180124_125441-B.jpg
 
----
 
Since T-Gel plain essentially contains no notable sugar, cellulose, starch and proteins, the main carbon source is phenols-other.
The mycelium produced new cells, stored sugars and other essential macro-micro nutrients, via the phenols-other.
 
The tea extract also provides the minimum essential macro-micro nutrients needed for growth.
Adding additives increases biological efficiency by 25-50% via various nutrients.
 
====
 
Notes:
 
Based on my experience with Cubensis Burma, I will need to transfer 3-4 times before best growth is observed.
The overall speed is slower than sugar-starch, as the process is longer and requires more oxygen.
 
----
 
Adapted Cubensis to T-Gel, second transfer (1,2). The third transfer produced white-rot (3).
 
IMG_20190107_130036.jpg IMG_20190115_215545.jpg IMG_20190413_113143.jpg
 
----
 
Single Cubensis T-Gel wedge to brown rice, first time decaying starch and grain.
 
IMG_20190205_133120.jpg IMG_20190208_202026.jpg IMG_20190208_202038.jpg
 
Tip: Mix and wait for re-colonization, 2-3 times.
 
----
 
Notice the oxidation of actives after spawning (black-blue):
 
IMG_20190212_183504.jpg IMG_20190212_183509.jpg
 
====
 
How I germinated the Cubensis spores:
 
First I cut down a wooden toothpick to the size I wanted using wire cutters, and pressure cooked it with sugar water (10%).
The peg was flamed over before use, to remove excess water and further cleaning, using metal tweezers.
 
Using the tip of the peg I collected spores from a print and plugged it into WL-Tek (general).
After 3 days the peg area was fed with additional sugar water (sucrose, syringe).
 
After 5-7 days the peg should be ready enough to transfer to agar-other.
 
IMG_20181104_193018.jpg IMG_20181022_192802.jpg IMG_20171116_170505.jpg
IMG_20181108_122512.jpg IMG_20181108_123034.jpg IMG_20181122_133646.jpg
IMG_20171122_142056.jpg IMG_20171124_132944_-_E.jpg IMG_20171203_140303.jpg
 
====
 
I forgot to mention, the first transfer for Cubensis on T-Gel was experimental, and included additional nutrients, it produced knots but no fruits:
 
IMG_20181222_124016.jpg IMG_20181222_124329.jpg IMG_20190101_115805.jpg IMG_20190101_120008.jpg IMG_20190101_120201.jpg
 
1.6g Black Tea (50%, 0.6g Soluble), 0.8g Sucrose, 0.04g MG (see here).
 
====
 
Azure to T-Gel, 1st transfer:
 
Slowly producing a radial, growth follows the preceding decay of enzymes-reactants.
Speed of growth is slightly slower than transfer 2 Cubensis to T-Gel.
 
IMG_20190506_143246.jpg IMG_20190506_143430.jpg
 
====
 
Azure sample to T-Gel, transfer 2 will have additional MG added to see if it speeds up growth via nutrients.
The enzymes-reactants needed to decay the phenols contain other elements (e.g. copper).
 
IMG_20190507_130323.jpg IMG_20190507_130426.jpg IMG_20190507_130513.jpg IMG_20190507_130556.jpg
 
In addition, proteins and other materials require nitrogen.

 


Edited by Ferather, 07 May 2019 - 07:30 AM.


#59 Ferather

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Posted 08 May 2019 - 08:36 AM

====
 
Agar:
 
Nearly all agar recipes used are sugar or starch based, T-Gel contains no sugar or starch (reduces contamination).
Many organisms die to oxygen (see my T-Gel recipe), phenols-other require oxidation to decay.
Lignicolous-other mycelium produce enzymes-reactants to oxidize materials.
 
 
Actives:
 
Active materials are a type of antioxidant, antioxidant's protect the mycelium from oxidative damage.
If the mycelium is oxidizing a substrate-material, it must protect it's self from decay.
 
----
 
Cubensis used substrate sample, fruiting in high O2. Enriched paper.
 
Cubensis-Fruits-2.jpg     955508151-PC_5.jpg
 
The second image was a decay test.
 
====
 
Today's images:
 
Growth is highly populated, linear, not rhizomorphic, it's vigorous but slow (the process-conversion takes longer).
The adaptation is nearly produced a radial, it's likely genetics-messages where passed around.
 
IMG_20190508_135620.jpg     IMG_20190508_135738.jpg     IMG_20190508_140233.jpg     IMG_20190508_140236.jpg
 
It's producing sugars-cells-other from the decay of phenols-other.


#60 Ferather

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Posted 09 May 2019 - 08:56 AM

The Azure appears to be more focused on population, decay and resource collection (and conversion), I also see possible varied genetics.
In comparison to Cubensis Burma on T-Gel, plain, population rate is about 75% higher, outgrowth is about 75% lower.
 
IMG_20190509_140120.jpg   IMG_20190509_140126.jpg       IMG_20190509_140225.jpg   IMG_20190509_140233.jpg
 
The surface should eventually be completely populated.





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