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Cubensis [Data Log]


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#1 Ferather

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Posted 28 January 2018 - 02:27 PM

I decided to try and put all of my Cubensis data into one post for better reading, I may update this thread over time.
Feel free to ask questions, correct or add relevant information in regards to Cubensis and growth.
 
Most of the information will be exports of previous posts, with editing to suit.
 
----
----
 
Cubensis does not expel laccase enzymes, and does not target phenol's as carbon, it requires starch-sugar instead (work around).
Dung has a pH of 7+, it also contains ammoniacal and ureic nitrogen, along side proteins and other nitrogen sources.
 
Dung is also composed of plant fiber, where all or most of the starch-sugar has been absorbed.
The dung will be cellulose based, plus other materials the animal cannot decay.
 
Cellulose as almost inert, non-reactive, and very slow to decay.
Decay is improved via an easier carbon source.
 
----
 
Mycelium like to oxidize substrates as part of their normal decay and feed routine.
Cubensis, when fed, can decay phenol's (untargeted) via oxidization.
 
----
 
Enzymes
 
Disclaimer: Optimal pH varies with the organism, and it's enzymes.
 
 
Cellulase (cellulose): 4.5-8.5.
Laccase (phenol's): 5.5-6.0. < Lignicolous mycelium.
Amylase (starch): 3.0-7.0.
 
Protease (protein): 2.5-11.
 
 
For cellulase there are four types:
 
Alkaline cellulase: 7.2-8.5.
Neutral cellulase: 6.0-8.0.
Hybrid cellulase: 4.5-7.0.
Acid cellulase: 4.5-5.0.
 
 
For amylase there are three types:
 
α-Amylase: 6.7-7.0. < Any amount of glucose.
β-Amylase: 4.0-5.0. < Cleaves two units.
γ-Amylase: 3.0. < Cleaves one unit.
 
 
Laccase remains stable at pH 5.0-10.0.
 
 
For protease there are three types:
 
Alkaline protease: 7.0-11.0.
Neutral protease: 6.0-9.0.
Acid protease: 2.5-4.0.
 
----
 
Proteins count both as a carbon source and as a nitrogen source.
 
----
 
Manure:
 
Contains: Ammoniacal and ureic nitrogen, P2O5, K2O plus other nutrients.
 
Manure.jpg
 
----
 
Fertilizer, such as MG:
 
Contains: Ammoniacal and ureic nitrogen, P2O5, K2O plus other nutrients.
 
IMG_20160718_170209.jpg IMG_20160718_170210.jpg
 
----
 
We know Cubensis expels cellulase, and likes pH 7 or higher. And what nitrogen it likes.
 
Conclusion: Paper pellets (no phenol's, pH 7-7.5) + nitrogen + starch spawn.
 
The same method also works on lignicolous mycelium.
 
----
 
You can refer to my T-Gel to see lignicolous mycelium decay and spend phenol's like they where easy sugars.
Lignicolous mycelium will germinate using phenol's, and spend the phenol's on cellulose-other.
 
----
 
Cubensis will also decay lignicolous materials if the pH is changed, and sugar-starch is added.
 
----
 
WL-Tek: Macro-micro nutrient enriched cellulose, pH 7-7.5.
 
 
Here is Cubensis spores on WL-Tek + wood peg with trace sucrose.
It germinated and grew after 4 days, but required sucrose.
 
IMG_20171116_170505.jpg IMG_20171117_185534.jpg
 
 
Here is Cubensis grain spawn to WL-Tek recycled paper.
 
955507891-PC_1.jpg 955507975-PC_2.jpg
955508050-PC_3.jpg 955508095-PC_4.jpg
 
 
It's better to enrich the paper with "carbon free" additives, to keep it clean.
Any extra carbon may be "available" to invader contamination.
 
----
 
Here I spent a tea bag in 250g CaCO3 water (pH 8.0), to remove the phenol's.
I added trace sucrose, because Cubensis is "not" lignicolous.
 
IMG_20171122_142056.jpg IMG_20171122_142204.jpg
 
It regenerated and grew out after only 8 hours.
 
IMG_20171122_142402.jpg IMG_20171122_180413.jpg
IMG_20171123_131246_-_E.jpg IMG_20171123_131352_-_E.jpg
IMG_20171124_132944_-_E.jpg IMG_20171125_214256_-_E.jpg
IMG_20171130_213226.jpg IMG_20171203_140303.jpg
 
The end pH was roughly 6.5.
 
----
 
Tea is rich in macro-micro nutrients, but required a carbon source to replace the phenol's.
 
Carbon, nitrogen and other nutrients: Millet, Brown Rice, Tea Leaves.
 
Tea leaves carbon sources: Phenol's, cellulose, proteins.
 
----
 
The science currently applies, I will explain. Most of you will be using grain and coir, maybe with vermiculite for more water-trace other.
Coir is composed of cellulose, phenol's (lignin-other) and trace proteins as carbon, carbon-nitrogen, but it needs treating.
 
 
"Because coir pith is high in sodium and potassium, it is treated before use as a growth medium for plants or fungi,
by soaking in a calcium buffering solution; most coir sold for growing purposes is said to be pre-treated." -- Source.
 
 
The coir you should be using will be more like paper pellets, and have a higher pH.
Ideally the coir should have a pH of 7-7.5 to mimic dung as a substrate.
 
pH 7.0 is also ideal for α-Amylase, the best starch enzyme.
 
----
 
So now you have coir, ready to use on fungi, it's high in cellulose and "should" only germinate molds on the trace phenol's.
You add spawn as normal, the mycelium will be spending the starch, and macro-micro nutrients on the coir.
 
Mycelium make substrates acidic over time, when α-Amylase destabilizes, cellulose takes over.
β-Amylase if far less efficient and releasing carbon and nutrients from the grain.
 
The mycelium must now rely on the coir for both carbon and nutrients.
 
----
 
Wood pulp (paper) and Acid-free paper. For me its miles cheaper than coir.
 
IMG_20160907_224246.jpg IMG_20170420_162701.jpg
 
----
 
Notes:
 
Coir is a modified lignicolous based material (like paper).
The nutrient additives in WL-Tek are "soluble".
 
----
 
Here is some data on acid hydrolysis of starch, unfortunately it's does not apply at room temperature much.
In case you where wondering if the acidity decays starch, it does, but it's not effective below 85°C.

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#2 Ferather

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Posted 30 January 2018 - 04:31 PM

Flow guide of enzymes and feeding:
 
----
 
Tip: Enzymes effect biological efficiency (no feed, no efficiency).
 
----
 
Nutritional media > enzymes > settings |> small-soluble units > units absorbed through cell wall > metabolize > produce materials > grow, do other.
 
Using the flow guide for specific materials:
 
Caffeine > laccase > settings |> ...
 
Settings explained:
 
There are various settings that ultimately change or stop enzyme performance.
Using the laccase example, it would become unstable below pH 5.
 
If the media pH is 4, the flow stops at "settings".
 
----
 
Next steps:
 
A nutrient additive your mycelium utilizes, due to deficiencies.
Know what ratio it wants, and the ratio of your substrate.
 
Example:
 
The media I am using is 500:1, and my mycelium does best on 25:1.
I will need to add nutrients to improve the total C:N ratio.
 
Example website here, composting (decay).
 
----
 
Cell walls are composed of proteins, chitin and glucans.
Both the proteins and chitin contain nitrogen.

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#3 Ferather

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Posted 30 January 2018 - 07:33 PM

Soil Amendments and Fertilizers (Some irrelevant info, but a good read).



#4 Ferather

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Posted 01 May 2018 - 08:07 AM

Cubensis also utilizes calcium carbonate (CaCO3) when it becomes soluble. Growth is more vigorous at pH 7-8 than at pH 5-6, this is likely due to lack of laccase, and inhibitory effects of phenols-other.

I conclude recycled paper pellets can be used in replacement of coir, in cases such as mine, where the paper pellets are cheaper and easier to use, they also have a higher pH of 7.

 

If you intend to use 10% or less spawn the paper will need to be enriched, this is because wood has a very low nitrogen content (the acidic phenols are also removed).

Spawn will be required to utilize cellulose, this is due to the slow release nature of the cellulose, where the starchy spawn accelerates decay.

 

Alternatively add starch or sugars to the cellulose, however, this will increase invader growth chances significantly.



#5 Ferather

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Posted 01 May 2018 - 08:10 AM

In addition: Coco coir is a lignicolous material (cellulose-lignin), which naturally has a high sodium and potassium content, and must be treated.

 

https://en.wikipedia.org/wiki/Coir


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#6 Microbe

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Posted 01 May 2018 - 11:53 AM



In addition: Coco coir is a lignicolous material (cellulose-lignin), which naturally has a high sodium and potassium content, and must be treated.

https://en.wikipedia.org/wiki/Coir


I didn't learn about this until i started growing plants using a coir based media. Using coir untreated has never been a issue for me. I want to use a premium coir that has been rinsed and treated. It even has some trichoderma in it but this shouldnt be a issue because of the pasteurization process. I know this seems like jumping into a pool that has down powered lines laying it and your pretty sure they are not charged but you never know until its too late.

I think EC is something i want to look into with my mushroom substrates. Here is a link that talks about it with coir and this is referencing washed coir.

https://www.maximumy...ng-media/2/1318


I bet the standard bricks we use that are intended for reptile bedding have a a ppm of salt near 1500. Salt is detrimental to mycelium and there is no doubt that you can probably establish better growth parameters with washed coir.

#7 Ferather

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Posted 01 May 2018 - 01:34 PM

I've seen people use untreated coir on certain cultures and it produce bad-odd-stalled growth. My guess is that some coir is not only untreated but also too fresh.

The higher salt would probably be beneficial as pet bedding, as it should deter mold, enough to make it last a bit longer (like bread).

 

Adding 10% CaCO3 to pH 7 paper produced pH 8.5, it increased BE (due to solubles) and reduced invader chances.

I managed about 4-6 months, using paper (WL-Tek) with 10% CaCO3 without invaders.

 

The same setup outdoors, grew algae after about 3-4 months.


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#8 Ferather

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Posted 04 November 2018 - 11:43 AM

This post is old, and has been updated in my journal (see my signature). I am continuing from this post, in order to not take over someone else's thread.

 

----

 

The Cubensis sample is now producing a radial around the peg, and slowly speeding up.
All growth is at the tip of the peg, and on the agar, attracted to the tea.
 
The Cubensis appears to be decaying and utilizing phenols.
 
----
 
Yesterday's images:
 
IMG_20181103_145816.jpg IMG_20181103_145816_E.jpg IMG_20181103_145947.jpg IMG_20181103_145947_E.jpg
 
Today's images:
 
IMG_20181104_184436.jpg IMG_20181104_184436_E.jpg IMG_20181104_184554.jpg IMG_20181104_184554_E.jpg

Edited by Ferather, 04 November 2018 - 02:22 PM.


#9 Ferather

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Posted 05 November 2018 - 10:31 AM

Well it's decaying the phenols, and also using them in replacement of sugars.
Growth is accelerating daily, and is highly populated (tomentose).
 
Growth on the peg is now increased due to the agar.
 
IMG_20181105_150907.jpg IMG_20181105_150907_E.jpg
IMG_20181105_151027.jpg IMG_20181105_151027_E.jpg
 
Typically short in length.
 
----
 
Notes:
 
The Cubensis is oxidizing the T-Gel as a lignicolous mycelium would, and is producing a dark ring.
Tea extract is nutritional like grain water (nitrogen, potassium, vitamins, and so on).
The mycelium, or capillary action, or both, is drawing up various solubles.
No simple sugar or starch is present in T-Gel agar (plain).
 
----
 
I will take further images in around 5 days for better observation and results.


#10 Ferather

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Posted 08 November 2018 - 08:06 AM

A little early, but here are some images. The Cubensis does decay the phenols, however it's less tolerant of the inhibitory effects than a lignicolous mycelium.

As decay of the cellulose (peg) improves and more sugars are released, the Cubensis is more able to tolerate and decay the inhibitory effects.

 

Decay of the phenols is also slower, but more rewarding than sugars (more carbon per gram).

I will try a modified T-Gel attempt, with added sucrose and essential nutrients.

 

IMG_20181108_122512.jpg IMG_20181108_123034.jpg



#11 PJammer24

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Posted 08 November 2018 - 11:09 AM

Just wanted to mention that most manures have PH levels of 7- and not 7+... It depends on the manure, some can be slightly higher than 7 but typically they are a little more acidic. Manure can have a PH level in the low to mid 4 range.

 

This is why it is sometimes beneficial to buffer the PH when using manure based substrates. I think for the most part, when mixed with typical substrate components like vermiculite and coir or straw which have a neutral PH, the buffer isn't necessary because when combined, the overall PH is favorable to the mycelium... At least with cubes. Cubes like PH levels that are neutral or slightly acidic. The combination of the manure with a PH level of less than 7 with two neutral additives like coir and verm, make for a comfortable growing environment for cubes. If you are working with manure that is at the low end in terms of PH level, like around the mid 4 range, the PH buffer becomes beneficial.

 

I kinda got off topic there and I apologize but my point is that the PH of poo can vary and I don't think 7+ as you indicate in your earlier post is accurate.



#12 Ferather

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Posted 08 November 2018 - 02:59 PM

It's accurate, the dung which Cubensis will fruit from (because it won't fruit from all dung) has a pH of 7+, or at least 6.5 or higher.

In addition, the pH of the manure can increase when composted, however you are correct with the mixtures.

 

Fig4_aop1113.jpg

 

Elephant dung pH test video here.

 

----

 

As mentioned this log is old, and has been updated (see my signature below).

You can say anything you like, I am 100% willing to accept corrections.

 

You can also read my CaCO3 test data in the same journal.



#13 PJammer24

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Posted 08 November 2018 - 03:44 PM

It's accurate, the dung which Cubensis will fruit from (because it won't fruit from all dung) has a pH of 7+, or at least 6.5 or higher.

In addition, the pH of the manure can increase when composted, however you are correct with the mixtures.

 

Fig4_aop1113.jpg

 

Elephant dung pH test video here.

 

----

 

As mentioned this log is old, and has been updated (see my signature below).

You can say anything you like, I am 100% willing to accept corrections.

 

You can also read my CaCO3 test data in the same journal.

 

This is from that same journal:

"Manure pH (except for poultry manure) reduced significantly with composting (Figure 4). The pH of buffalo manure was 8.7 in fresh and reduced to 7.7 in composted sample. For camel manure, the pH was 8.6 in fresh and 8.5 in compost, for cow manure pH was 8.5 in fresh and 7.4 in manure compost. Goat fresh manure showed pH of 8.9 and 7.8 in composted manure. Poultry manure had a pH of 8 in fresh and 8.4 in compost. This decrease in pH could be attributed to the modified chemical composition of manure through microbial action, notably the production of organic acids."

 

 

 

it say right here in the paragraph above the bar graph you posted that and I quote "Manure PH (except for poultry manure) reduced significantly with composting" 

 

Then there is a bar graph that shows that the PH increased with composting... I have a hard time taking as fact things stated in a journal that has a bar graph which contradicts the previous paragraph on the same topic.... Or am I missing something...???

 

 

 

That being said, The information I was posting was for Horse manure which is not on the chart you provided. It is my understanding that horse manure is the best substrate for the mushroom in question... Following is a little more data that I pulled from various sites asserting that Horse manure has a PH of 7 or below. (This is just data from the internet, please bear with me, I will test the real stuff at home when I get a chance.)

 

Horse Manure Compost: The Benefits

  • Fully organic soil improver
  • Increases the level of rich organic material in your soil
  • Increases the level of nutrients in your soil
  • Perfect for mulching roses
  • Contains no pests or pathogens
  • Reduced odour

Product Composition

  • Well-rotted horse manure
  • pH Level: 5.83
  • Ammonia: 374 mg/kg
  • Calcium: 37152 mg/kg
  • Copper: 71 mg/kg
  • Dry Matter: 24.7%
  • Magnesium: 0.34%
  • Nitrate: <0.1 mg/kg
  • Phosphorus: 0.51%
  • Potassium: 0.84%
  • Sodium: 0.15%
  • Sulphur: 0.32%
  • Zinc: 120 mg/kg
  •  

 

 

 

"Adding pure, fresh horse manure will make your soil more acid. 
Adding well-rotted pure horse manure - less acid, but depending on whether the muck was covered up/outdoors, some of the nutrients, especially Nitrogen, may have leached out / evaporated. 
Fresh manure with straw : straw [high Carbon] will have the effect of 'locking up' some of the Nitrogen until the straw breaks down, when you get a more stable [hohoho] form of N, which is a good thing - slower release for plants. 

Well-rotted manure with straw - straw should have broken down at least partially [see immediately above] 

Adding manure will always make your soil more acid. "

https://forum.downsi...4f7a13d9b2476fd

This is just from a horticulture message board and has less validity in my opinion though it supports my assertion so I will include it.

 

 

 

"The pH of peat moss is around 4, making it a suitable soil amendment for plants that thrive in acid soil. Aged manure averages somewhat higher: from 4.6 to 7.4, depending on the type of manure."

https://homeguides.s...nure-37732.html

 

 

"Most sources report the pH of compost between 6 and 8, without specifying a source for this information. Reference 1  shows the pH of various composts and most are between 7 and 8. For example: yard debris 7.7, mixed manure 7.9 and leaf 7.2. The ones below 7 include horse manure at 6.4 and bark compost at 5.4. Reference 2 studied home composts and found a mean pH of 7.0 to 7.5. "

https://www.gardenmy...es-acidic-soil/

 

 

"Another possibility is to test the pH of the horse’s fresh manure, using a “food probe” pH meter. Normal pH is about 6.8, and anything at 6.5 or below is indicative of hindgut acidosis and the circumstances for colonic ulcers to form."

http://www.succeed-e...-in-your-horse/

 

 

"The pH in the hindgut is generally 6.5-7"

https://www.hygain.c...osis-in-horses/

 

 

I can attest to these figures in that horse manure has a ph of around 7 or less from having horses who are suffering from colic and having the condition diagnosed by vets.

 

If the chart you provide IS accurate, using any of those manures means that certainly no calcium type additives should be used in the substrate to buffer the PH. If anything, The PH should be lowered when using any of the manures detailed in the bar chart you provided. IF THAT INFO IS CORRECT

 

Horse manure, which is what I have worked with most often (I have worked with cow also) is considered the most appropriate manure to be using in this hobby, at least that is my understanding. Of the manures in question it is the lowest in terms of PH. My assertion was specific to the manure I have worked with and I should not have spoken so generally. Perhaps certain manures are more appropriate for certain types of mushrooms. I might be mistaken, but I believe commercial Agaricus are grown using cow manure. Perhaps they appreciate a more alkaline growing substrate, I would have to look as I have not attempted to grow the finicky little bastards. 

 

If I get around to it, i will test the PH of the raw manure and the compost I have sitting around and see what we come up with.

 

compost
 
The data I posted is from sites on the net... i will try to find time to acquire some data on this personally rather than believe what is on the internet... I know I know right... I am spinning my wheels for no reason... Everything posted on the internet is TRUE... 

Edited by PJammer24, 08 November 2018 - 04:04 PM.

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#14 Cuboid

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Posted 08 November 2018 - 04:19 PM

Check the units on the vertical axis of the bar graph. It's not pH directly. It's conductivity. Not sure exactly how that translates to pH value. Presumably there is an inverse relationship though.
Edit: After a quick Gulag search I conclude there is no general relationship between pH and electrical conductivity of soil/manure.

Edited by Cuboid, 08 November 2018 - 04:29 PM.


#15 Ferather

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Posted 08 November 2018 - 04:33 PM

That's quite a bit of data. I'm glad you are willing to include science and varied data sources (plus testing). I'm not saying don't add lime, since I add lime to my substrates (various reasons, and purposes).

However at pH 4.5 Cubensis stalled, at pH 6.5 (acids removed) it grew but fairly slowly. Acidic phenols and and other acids can be inhibitory, when removed growth improved.

 

Various acids can breakdown to alkali's, decay overtime, be saturated in water, removed (washed away), or diluted prior to use (as you mentioned).

I can germinate Cubensis on cellulose + essential macro-micro nutrients at pH 7.5-7.8, with lime seems to be slightly preferred.

 

In my journal I placed a CaCO3 barrier onto brown rice, which later I transferred to unmodified paper (test).

There is much more info in my Journal (Including enzymes, which are effected by pH).

 

996533651-Cubensis-CaCO3-1.jpg 996534178-Cubensis-CaCO3-2.jpg 996534562-Cubensis-CaCO3-3.jpg 996534653-Cubensis-CaCO3-4.jpg 996535041-Cubensis-CaCO3-5.jpg 996535428-Cubensis-CaCO3-6.jpg 996535791-Cubensis-CaCO3-7.jpg 996536199-Cubensis-CaCO3-8.jpg 996536294-Cubensis-CaCO3-9.jpg

 

At pH 7-7.5, decay of cellulose and starch via enzymes was improved.

 

----

 

Acidification (Simple edition):
 
Acidification can cause mycelium to fruit (various reasons), allowing for mild acidification and then increasing the pH, increases the number of cycles.
 
Once the media pH reaches a level the mycelium and-or it's enzymes are no longer stable, it will not produce any more fruits.
At this point the media pH must be increase (reset to original), any waste potential should also be removed.
 
This is where 'some' secondary decomposers would come in, tolerant to the acidification (acid stable enzymes and low pH tolerance).
The dung which cubensis likes, is not acidic, it has a pH of 6.5+, so some decomposers come in when the pH has increased.
 
As acidification takes place, and enzymes begin to destabilize, mycelium will start to respond to low(er) nutrients or low(er) uptake.
Usually there a few different types of enzyme that target a material, the main setting difference is the stable pH range.
 
Acidification can also effect various protease enzymes, but not other enzymes (causing a low nitrogen uptake).

 

----

 

I hope we can agree on 6.5+ for starting a colony, I accept corrections.


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#16 PJammer24

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Posted 08 November 2018 - 04:35 PM

Ahhh, Now I'm a little embarrassed that I did not fully read the journal either...  :tinfoil:   :blush:  :blush:  Thank you Cuboid... Admittedly, I know nothing of the electrical conductivity of soils and how that impacts growth or PH.



#17 PJammer24

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Posted 08 November 2018 - 04:37 PM

I really should be working right now...  I don't feel like searching for sources at the moment.... 

 

But

 

It is my understanding that the strain in question, cubensis, grow best at a neutral PH or slightly less that neutral....


Edited by PJammer24, 08 November 2018 - 04:37 PM.


#18 PJammer24

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Posted 08 November 2018 - 04:38 PM

So yes, we can agree on that!! Cheerio mate!!


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#19 Ferather

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Posted 08 November 2018 - 04:42 PM

@Cuboid

 

Good spot, no idea why that came up in the search (that's what I get for trying to get a combined image).

 

https://patents.goog...2017140947A1/en (Image near the bottom).

 

----

 

@PJammer24

 

No problem, let me know if you have any questions, or think anything is wrong.

Thanks for you input, I am taking notes from your pH tests.


Edited by Ferather, 08 November 2018 - 04:44 PM.


#20 Ferather

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Posted 09 November 2018 - 07:27 AM

I have modified-updated yesterdays post (another forum)
 
----
 
Here are yesterdays images (08-11-18). The Cubensis does decay the phenols, however it's less tolerant of the inhibitory effects than a lignicolous mycelium.
As decay of the cellulose (peg) improves and more sugars are released, the Cubensis is more able to tolerate various inhibitory effects.
 
Decay of the phenols is much slower than sugars, but also more rewarding than sucrose (more carbon per gram).
I will try a modified T-Gel attempt, with added sucrose and soluble essential nutrients.
 
The yellowing at the tip is either the light or my camera.
 
IMG_20181108_122512.jpg IMG_20181108_123034.jpg
 
----
 
Additional data drawn from Mycolorado's T-Gel with Malt extract (image here).
 
Note: I may need to alter the end pH of the T-Gel.
 
----
----
 
Todays images, looks like a Cubensis tree (overall shape), which is fairly funny based on the composition (and test).
Growth on the peg is continuing at a good speed, growth on the T-Gel is now extremely slow.
 
IMG_20181109_115357.jpg IMG_20181109_115529.jpg
 
I will transfer soon.
 
----
 
Mycolorado's alternative T-Gel recipe: 100g > Water | 2g > Agar | 2g > Black Tea | 1g > Malt Extract.

Edited by Ferather, 09 November 2018 - 07:58 AM.





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