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Cubensis [Data Log]


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#21 Ferather

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Posted 10 November 2018 - 09:55 AM

Some further thoughts, as I was looking at the black oxidation ring (tea phenols turn black when they decay to oxygen).
I may need to increase GE (oxygen intake), more than normal (for example sugars), for improved decay.
 
I cannot conclude without a LAB, what types of laccase and-or peroxidases Cubensis uses.
 
----
 
Next test: 100g Water, 2.4g Agar, 1.6g Black tea, 0.8g Sucrose, 0.04g MG.
 
Note: Around 25-33% of the tea is soluble, not all of it.
 
 
Nutrients:
 
 
 
Proteins, varies:  [ Cx | Hx | Ox | Nx ] x. -- Trace, minimal.
Polyphenols, varies:  [ C6 | H5 | O ] x. -- Phenols.
Cellulose: [ C6 | H10 | O5 ] x. -- Trace, debris.
 
Theobromine:  C7 | H8 | N4 | O2. -- Phenol.
Theophylline:  C7 | H8 | N4 | O2. -- Phenol.
Caffeine:  C8 | H10 | N4 | O2. -- Phenol.
Theaflavin:  C29 | H24 | O12. -- Phenol.
Tannins:  C76 | H52 | O46. -- Phenol.
Catechin:  C15 | H14 | O6. -- Phenol.
 
Carotene:  C40 | H56. -- Hydrocarbon.
 
 


#22 Ferather

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Posted 11 November 2018 - 09:17 AM

I transferred the peg yesterday (24 hours ago), I forgot to wipe the inside of the lid after sterilizing it.
Regardless, the Cubensis has recovered and started growing out (also on the agar).
 
The black oxidation ring has already appeared due to active enzymes.
 
IMG_20181111_124111.jpg IMG_20181111_124208.jpg IMG_20181111_124228.jpg
 
Now with more GE, more oxygen.


#23 Ferather

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Posted 12 November 2018 - 04:03 PM

[Recipe B + More GE]
 
48 hours: Speed, decay rate and morphology have all changed.
 
Oxidation of the phenols is more obvious (black ring), speed has improved, the peg is still producing growth.
Morphology of the mycelium on the agar is no longer cottony, but instead it's flat and linear.
 
Damage from transfer is almost gone, peg growth appears more pushed out.
 
IMG_20181112_192529.jpg IMG_20181112_193112.jpg

----
 
Just noticed a green-blue-black tint, in various areas on the peg, just about visible in the images.
I guess increased nitrogen, and-or increased nitrogen, phosphorus and oxygen?
 
----
 
Baeocystin:  [C11]--[H15]--[N2]--[O4]--[P]
Norbaeocystin: [C10]--[H13]--[N2]--[O4]--[P]
 
Psilocybin:  [C12]--[H17]--[N2]--[O4]--[P]
Psilocin:  [C12]--[H16]--[N2]--[O]
 
 
From above:
 
"Psilocybin is a tryptamine compound with a chemical structure containing an indole ring linked to an ethylamine substituent. It is chemically related to the amino acid tryptophan, and is structurally similar to the neurotransmitter serotonin."
"Psilocybin is a member of the general class of 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"
"Psilocin is broken down by the enzyme monoamine oxidase. Some psilocin is not broken down, and forms a glucuronide"
 
 
Notes:
 
Nor-baeocystin are analogs of psilocybin, meaning single or various elements are added or removed.
It appears they can be phosphorylated or dephosphorylated (cleaved, added, a cycle?).
All four compounds contain carbon, hydrogen, nitrogen and oxygen.
 
 
Serotonin:  [C10]--[H12]--[N2]--[O]

Edited by Ferather, 12 November 2018 - 07:20 PM.


#24 Ferather

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Posted 13 November 2018 - 06:12 PM

Today's image, with a modified image to better see the oxidation.
Growth is generally flat, linear, and also populated.
 
IMG_20181113_222310.jpg IMG_20181113_222310_E.jpg


#25 Ferather

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Posted 14 November 2018 - 05:58 PM

Today's image, I conclude Cubensis has the required enzymes to decay cellulose and phenols.
I therefore would take note of the various nutrients and ratios in dung, and it's pH.
 
This makes Cubensis a primary decomposer, but niche (N%, pH or both).
Cubensis also produces various types of protease enzymes.
 
IMG_20181114_221037.jpg
 
----
 
I suggest using 0.5-2g of normal additive with T-Gel, such as malt extract.
 
----
 
T-Gel (plain) and pH modification (not this test):
 
Increasing the pH to ~7, caused 3 types of mycelium to germinate.
The higher pH likely causes the phenols to break down.
 
I got pin mold, trich (green), and a random.
 
----
 
[More images soon]

Edited by Ferather, 14 November 2018 - 08:00 PM.


#26 Ferather

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Posted 15 November 2018 - 07:24 AM

15/11/18 | Morning
 
Visible adaptation, growing towards oxygen, high population rate.
Not the fastest speed, however growth looks robust.
 
IMG_20181115_115217.jpg IMG_20181115_115351.jpg


#27 Ferather

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Posted 17 November 2018 - 11:03 AM

Working as intended, not stalling. My camera, torch and lid are making image quality lower (sorry).
 
Growth is generally uniform, no visible rhizomorphs or overly cottony growth.
Best described as flat(ish), linear and generally populated.
 
IMG_20181117_153424.jpg IMG_20181117_153602.jpg
 
Might need a bit more MG.


#28 Ferather

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Posted 18 November 2018 - 06:42 PM

My Cubensis sample has changed a bit, due to the lid removal and lid change (more O2).
The new growth appears to be absorbing more phenols (slightly yellower).
 
I am happy with the results, although growth is a bit slow.
 
IMG_20181118_232405.jpg IMG_20181118_232502.jpg


#29 Ferather

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Posted 19 November 2018 - 04:27 PM

19/11/18 | Evening
 
It now looks like there are knots forming on the peg, however I think the peg is too dry for them.
Images 2 and 3 have some blueing, apparently there is a lot of oxidizing going on.
 
The pH, and acidity buildup might also be causing mycelial damage.
 
IMG_20181119_204132.jpg IMG_20181119_204221.jpg
IMG_20181119_204901.jpg IMG_20181119_204934.jpg


#30 Ferather

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Posted 22 November 2018 - 10:12 AM

Lignicolous Cubensis (which can be seen fruiting from "enriched" soils and composts), images:
 
T-Gel, test recipe (minimum sugars, sucrose, additional macro-micro nutrients):
 
IMG_20181111_124228.jpg IMG_20181122_133240.jpg IMG_20181122_133646.jpg
 
Spent tea bag (minimum sugars, sucrose, spent until roughly pH 6.5):
 
IMG_20171122_142056.jpg IMG_20171130_213226.jpg IMG_20171203_140116.jpg
 
In both cases the spores where germinated on WL-Tek, using a wooden peg with minimum sugars.
The pegs where later transferred to a lignicolous material, in this case black tea leaves.
 
Cubensis produces enzymes to decay cellulose, proteins, and phenols.
 
----
 
Inhibitory effects and lower pH's (acidic) effect general growth speed negatively.
Good growth was observed when additional nitrogen was added.
 
Tea leaves contain up to 3% nitrogen, none added.
 
----
 
Illustrations of what type of visible growth to look for:
 
IMG_20161009_183512.jpg IMG_20161009_184151.jpg
IMG_20161009_184506.jpg IMG_20161009_184555.jpg
955507891-PC_1.jpg 955507975-PC_2.jpg
 
Can be bushy when pushing up, or when populated.
 
955508050-PC_3.jpg 955508095-PC_4.jpg
996535041-Cubensis-CaCO3-5.jpg 996535791-Cubensis-CaCO3-7.jpg
 
Yes it's growing on CaCO3 rocks.

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#31 Ferather

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Posted 29 November 2018 - 08:41 AM

WL-Tek test: 112g Water, 25g Paper Pellets, 12.5g Black Tea Leaves, 6g Ultra Fine CaCO3, 1g MG, 0.1g YN.
 
I started with 160g of water, it was then cooked down to 112g (~72% water), using a microwave.
The water was added to the additives, mixed, then added to the pellets.
 
----
 
T-Gel test: 100g Water, 2.4g Agar, 1.6g Black Tea, 0.04g MG, 0.02g YN  |  No sugar, or flour.
 
I transferred a wedge 12 hours ago, and have visible growth already.
It was relocated it after the drop (hence the left patch).
 
314867889-IMG_20181125_115425.jpg 349204190-IMG_20181129_112006.jpg
 
----
----
 
Now +6.5 hours from the previous image, and it's a personal record.
Probably the nutrient combination (MG + YN) + phenols.

 

351770545-IMG_20181129_184327.jpg 351771396-IMG_20181129_184422.jpg


Edited by Ferather, 29 November 2018 - 02:03 PM.


#32 Ferather

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Posted 30 November 2018 - 05:55 PM

Growth is faster, thicker and very populated, carbon (phenol) utilization has increased with additional macro-micro nutrients.
Current speed is faster than Tarragon oyster on T-Gel (plain, no flour) but enriched (MG @ 0.1g, no YN).
 
Even the mycelium left from the wedge relocation has regenerated and is growing out.
The wedge now appears to be turning darker (phenols oxidizing).
 
IMG_20181130_215309.jpg IMG_20181130_215436.jpg
 
----
 
MG-YN.jpg
MG.png

Edited by Ferather, 30 November 2018 - 06:29 PM.


#33 Ferather

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Posted 01 December 2018 - 06:07 PM

Working as expected, and it's fast, the added array of macro-micro nutrients combined with the carbon rich phenols makes a viable media.
This suggests enzymes and-or reactions needed to decay and utilize the carbon source (phenols) have increased.
 
Note: There is almost no simple sugar (0.012%, 100g of T-Gel, from tea) and no starches.
 
I conclude Cubensis can and will target phenols for decay.
 
IMG_20181201_212951.jpg IMG_20181201_213543.jpg
 
----
 
Now to focus more on grow medias.


#34 Ferather

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Posted 02 December 2018 - 09:41 AM

02/12/2018 - 14:30 (+15 hours)
 
The WL-Tek test is working, the phenols are speeding up decay. However, more of the phenols are being spent on additional enzymes-other.
Instead of just decaying phenols and growing out, more resources are being spent on decaying other materials (slower).
 
In addition, the soluble nutrients are more locked into the paper cellulose, and less free.
The wedge is a little close to the lid, and producing condensate.
 
IMG_20181202_134307.jpg IMG_20181202_135901.jpg


#35 Ferather

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Posted 03 December 2018 - 04:44 PM

The next test I will make sure the wedge is not too close to the lid, sorry for the images.
Regardless, I have lignicolous Cubensis on WL-Tek (cellulose) + tea.
 
IMG_20181203_210012.jpg IMG_20181203_211227.jpg
 
Speed is improving.


#36 Ferather

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Posted 05 December 2018 - 06:33 PM

Growth is very slow, spending it's time and resources on slow release cellulose.
This time it's not stalling like the 100% cellulose sample did.
 
Not as fast as WL-Tek + 8g sucrose.
 
IMG_20181205_230316.jpg IMG_20181205_225613.jpg

Edited by Ferather, 05 December 2018 - 06:37 PM.


#37 Ferather

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Posted 06 December 2018 - 10:52 AM

WL-Tek + Tea, +16 hours, with visible growth progression. T-Gel + MG-YN growth is weak.
Evidently, the Cubensis sample grows better with cellulose than without.
 
Decay of the phenols appears to be partial, not complete.
The sucrose in T-Gel test 1 improved growth.
 
IMG_20181206_153730.jpg IMG_20181206_153543.jpg





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