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OfflineFerather
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Testing antibacterial starch free T-Gel
    #23770605 - 10/25/16 04:28 PM (7 years, 3 months ago)

Decided to join katbusa at testing ways to make antibacterial agar mixtures.
I will be using sugar free jelly, so I can make a test run tomorrow.

So far katbusa has been successful with tea slurry agar.
I intended to test additional additives, and ratios.

And yes I have a powerful torch.


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Edited by Ferather (11/10/16 04:02 PM)


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Offlinekatbusa
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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23771157 - 10/25/16 07:23 PM (7 years, 3 months ago)

I will be writing up my tea agar methodology for you soon. Ive been super busy the last few days. Now that I think about it I might do it now since Im being lazy and sitting on my couch.


Edited by katbusa (10/25/16 07:23 PM)


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OfflineFerather
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Re: Testing antibacterial starch free agars [Re: katbusa]
    #23772285 - 10/26/16 04:06 AM (7 years, 3 months ago)

Just read it, great writeup so far. And I'm joining in the fun.
Everyone feel free to join in, post here if you like.


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OfflineFerather
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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23790569 - 11/01/16 07:38 AM (7 years, 2 months ago)

Triple test, I am using extra strong gelatin, and tea water. No starch. Open air assembly.

Test A, will I get an infection from the air on this composition.
Test B, will the gelatine turn to liquid, most probably.
Test C, will the mycelium expand and grow.

   

Set like silicone, hard to penetrate.


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OfflineMattisfat
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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23790603 - 11/01/16 08:01 AM (7 years, 2 months ago)

what you need to do is get a bacteria culture on agar - swab that onto a standard agar recipe and then swap it onto the tea recipe so you can see the difference in growth side by side.

maybe drop some mycelium in each container aswell to see how that does next to the bacteria.

Too many variables in your current way to get a clear view of the difference.

Currently im trying 10g black tea 9g malt extract 9g agar 500 ml water as im struggling with bacteria contams atm hopefully tea might help.


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OfflineFerather
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Re: Testing antibacterial starch free agars [Re: Mattisfat]
    #23790925 - 11/01/16 10:33 AM (7 years, 2 months ago)

Yea this one is semi un-serious due to the high chance the gelatin will turn to liquid.
Decided since that is going to do that, may as well test a few other things.

You are right though, just using up this gelatin.


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OfflineFerather
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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23791750 - 11/01/16 03:33 PM (7 years, 2 months ago)

4 hours, the gelatin seems to liquefy (digestion) then set again.

   

Mycelial debris growing, too fast for spores.
Will most likely end with a liquid.


--------------------
                   

Growing mushrooms, general guide and information (Ferather's Journal), https://ibb.co/rG3rML2

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OfflineFerather
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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23793835 - 11/02/16 07:13 AM (7 years, 2 months ago)

Turns out the liquefying was bacteria, the good news is, it has pretty much stalled.
The mycelium from the grain is growing out roughly at normal speed.

No obvious liquefying by any of the mycelium either.

Seems to be working, Images soon.


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OfflineFerather
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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23793848 - 11/02/16 07:25 AM (7 years, 2 months ago)

Side note:

No infection from open air assembly, most likely due to 0% starch and sugars.
This particular recipe will likely only work with live transfers.


--------------------
                   

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OfflineFerather
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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23800794 - 11/04/16 02:21 PM (7 years, 2 months ago)

Well, it works as intended, bacteria stalled after 24 hours, the mycelium won by day 4.
Not a single infection from open air assembly, possibly got lucky.

The strong gelatin is working 100% like agar.

   

No mycelial liquefying.


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OfflineFerather
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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23800828 - 11/04/16 02:34 PM (7 years, 2 months ago)

P.S. this was microwaved for 88 seconds then left to set, no pressure cooker.


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InvisibleMarty Mycfly
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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23800951 - 11/04/16 03:26 PM (7 years, 2 months ago)

Wow, that's pretty interesting! Only 88 seconds in the microwave?


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Mycfly's King Oysters                                       

Mycfly's Reishi    

                                                 


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OfflineFerather
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Re: Testing antibacterial starch free agars [Re: Marty Mycfly]
    #23801181 - 11/04/16 04:48 PM (7 years, 2 months ago)

Well not 100% true, for the whole recipe, which is as follows:

> 20g extra strong gelatin, 12g tea.
> 500g freshly boiled water.

Stew the tea for 5 minutes using a spoon, squeezing the bags.
Microwave for 2 minutes, and repeat again one more time.

Remove the bags and then microwave for 2 minutes.
Add the gelatin and microwave for 88 seconds.

Use as normal and leave to set, covered.


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OfflineFerather
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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23805789 - 11/06/16 09:08 AM (7 years, 2 months ago)

Today, Trich, under attack from King oyster.

   

The trich is weak, and cut off.


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OfflineQuadman
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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23805916 - 11/06/16 09:50 AM (7 years, 2 months ago)

Weak and cut off, but unfortunately still there.


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OfflineFerather
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Re: Testing antibacterial starch free agars [Re: Quadman]
    #23805939 - 11/06/16 09:59 AM (7 years, 2 months ago)

If the King destroys or outgrows it, it may inhibit it enough to prevent further growth.
That would allow me to continue growing and isolate a clean sample.


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OfflineQuadman
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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23806034 - 11/06/16 10:32 AM (7 years, 2 months ago)

If you want clean I'd be transferring from the opposite side now. That mean green will be all over with more time. You can let it grow out after the transfer for experimental purposes.


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OfflineFerather
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Re: Testing antibacterial starch free agars [Re: Quadman]
    #23806052 - 11/06/16 10:40 AM (7 years, 2 months ago)

I have stupid amounts of spawn, this is just a test. I isolated a sample and left it out for 8 days.
Got both bacteria and mold, the bacteria may well have already been there though.

Good enough for testing, not going to buy bacteria or make mold.


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Growing mushrooms, general guide and information (Ferather's Journal), https://ibb.co/rG3rML2

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OfflineFerather
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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23808892 - 11/07/16 08:35 AM (7 years, 2 months ago)

Well, the King is eating the trich for dinner.

   

The trich has also receded.


--------------------
                   

Growing mushrooms, general guide and information (Ferather's Journal), https://ibb.co/rG3rML2

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OfflineFerather
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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23808945 - 11/07/16 08:58 AM (7 years, 2 months ago)

Name of recipe: T-Gel Tek, read my copyrights.


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OfflineFerather
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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23809304 - 11/07/16 10:50 AM (7 years, 2 months ago)

Here is something I will attempt this evening or tomorrow. Using cocktail sticks (random link).
All I do is cook them and pin the Gelatin. When ready transfer to T-Gel or Agar asap.


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OfflineFerather
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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23809586 - 11/07/16 12:35 PM (7 years, 2 months ago)

The cocktail sticks took roughly 10-15 minutes to sink, not very long. Left to cool to room temperature.
Simple Guide: Cut or snap the sticks, boil some water, stew the sticks in a container with lid.

I will be testing fertilized T-Gel to see the results of added nutrients.
Also, I recycled the tea bag, and cooked it in grain water.


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OfflineFerather
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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23809776 - 11/07/16 01:40 PM (7 years, 2 months ago)

So I pegged my T-Gel carefully in a SAB, It took seconds for the cling film to attract dust after removing.
I used sterilized metal tweezers to remove the pegs and place them into my T-Gel sample.

In a few days I can attempt another transfer to T-Gel.

   

These are made from birch wood.


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OfflineFerather
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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23811908 - 11/08/16 07:46 AM (7 years, 2 months ago)

18 hours later, it has started to grow on the birch wood pegs.
When ready I will test a transfer to enriched T-Gel.

   


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Growing mushrooms, general guide and information (Ferather's Journal), https://ibb.co/rG3rML2

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OfflineFerather
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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23815472 - 11/09/16 08:06 AM (7 years, 2 months ago)

I will be able to transfer this evening, or tomorrow.

   

The trich suffers greatly in high CO2.


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Invisibleflyontoast
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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23816367 - 11/09/16 02:20 PM (7 years, 2 months ago)

Not fully catching the purpose to the sticks. You are letting the sticks get colonized and then are going to transfer the stick to a new dish rather than cut-out a wedge?

Separate question: how long can you leave myc on gelatin for? I was reading-up on pectin-"agar" and most people were saying that the myc digests the gelatin and the plate turns into a puddle. Is that not your experience, or is it true but you just don't leave the myc on one plate long enough for that to be a problem?


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:mushroom: My trade list :mushroom:
Looking for strong terrestrial fruiters for an outdoor beds experiment:
Agaricus Bitorquis, Agaricus Augustus, Agaricus blazei/subrufescens, Stropharia Rugoso-annulata, Clitocybe Nuda (blewits), and any species or other genus that you think work outdoors.
Also, any commercially viable Pleurotus, cold or hot strains.
Thanks for the Q&A, trades, and all the posters & teachers that have come before us  :skol:


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OfflineFerather
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Re: Testing antibacterial starch free agars [Re: flyontoast]
    #23816511 - 11/09/16 03:02 PM (7 years, 2 months ago)

The sticks are being used to transfer, correct. There are three types of Gelatin: Acid, alkaline or water extracted.
So far the strong Gelatin (acid) has stayed whole, no obvious mycelial liquefaction, still firm.

One transferred, The sample longer is no longer required.
The purpose is to clean, isolate and transfer.


--------------------
                   

Growing mushrooms, general guide and information (Ferather's Journal), https://ibb.co/rG3rML2

https://www.shroomery.org/forums/showflat.php/Number/27857366#27857366

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Invisibleflyontoast
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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23817416 - 11/09/16 07:34 PM (7 years, 2 months ago)


Did you come-up with this stick idea or was it already a tek? It's a brilliants device for making deep-dish and pre-poured agar plates easily workable,and all you need is tweezers! These colonized sticks would probably makes great slants too. I dunno, but when you combine pre-poured "agar" dishes, without the need for a scalpel, and less paranoia about bacteria, it really makes working with and storing plates a super low bar of entry for noobs. Not to mention yanking a handful of sticks out a plate to dump into grain has got to take way less time than making that many wedges.
I'm PCing a pre-poured agar now and am going to jam all sorts of toothpicks and chopsticks and whatever other sticks I have into it.


--------------------
:mushroom: My trade list :mushroom:
Looking for strong terrestrial fruiters for an outdoor beds experiment:
Agaricus Bitorquis, Agaricus Augustus, Agaricus blazei/subrufescens, Stropharia Rugoso-annulata, Clitocybe Nuda (blewits), and any species or other genus that you think work outdoors.
Also, any commercially viable Pleurotus, cold or hot strains.
Thanks for the Q&A, trades, and all the posters & teachers that have come before us  :skol:


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OfflineFerather
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Re: Testing antibacterial starch free agars [Re: flyontoast]
    #23818684 - 11/10/16 08:14 AM (7 years, 2 months ago)

Indeed flyontoast, the easiest transfer I have ever done, by far. It took about 45 seconds.

   

If this was a super rare wild clone, I now have a clean strong sample.
And yes this tek was my idea. It is free for all to use.


--------------------
                   

Growing mushrooms, general guide and information (Ferather's Journal), https://ibb.co/rG3rML2

https://www.shroomery.org/forums/showflat.php/Number/27857366#27857366

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Offline0yster
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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23819752 - 11/10/16 12:57 PM (7 years, 2 months ago)

:takingnotes:


Edited by 0yster (10/23/18 07:57 AM)


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OfflineFerather
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Re: Testing antibacterial starch free agars [Re: 0yster]
    #23819874 - 11/10/16 01:43 PM (7 years, 2 months ago)

After this test I will be transferring Black Poplar spawn to T-Gel.
Depending on test results, either normal or enriched.


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InvisibleMycolorado
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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23820020 - 11/10/16 02:46 PM (7 years, 2 months ago)

Quote:

Ferather said:
Indeed flyontoast, the easiest transfer I have ever done, by far. It took about 45 seconds.

   

If this was a super rare wild clone, I now have a clean strong sample.
And yes this tek was my idea. It is free for all to use.



That's a thing of beauty, Billy!


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OfflineFerather
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Re: Testing antibacterial starch free agars [Re: Mycolorado]
    #23820045 - 11/10/16 02:56 PM (7 years, 2 months ago)

If I did not have trich, I could have continued with open air assembly throughout.
However, the trich has not passed on the transfer, and I am successful.

My intention now is to test a variety of cultivated mycelium.
When complete I will post the T-Gel tek in full.

Any testing and input is welcome.


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Growing mushrooms, general guide and information (Ferather's Journal), https://ibb.co/rG3rML2

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OfflineFerather
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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23822283 - 11/11/16 08:31 AM (7 years, 2 months ago)

Here is 24 hours after the birch peg transfer.

   


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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23822813 - 11/11/16 12:04 PM (7 years, 2 months ago)

:pirate: You are a pirate Ferather! Good work!


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Re: Testing antibacterial starch free agars [Re: Marty Mycfly]
    #23823565 - 11/11/16 04:55 PM (7 years, 2 months ago)

Well the only thing to do now, other than side experiments, is test various mycelium and clean them.
If I can turn dirty spawn clean, I can clean anything that likes T-Gel, or adjusted T-Gel.

That means I can clone wild mushrooms, clean dirty vendor spawn, and so on.
Mycolorado tested several types of spores, most failed to germinate.

However that was T-Agar and not T-Gel, inconclusive.


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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23825472 - 11/12/16 10:22 AM (7 years, 2 months ago)

Here is 50% less tea but enriched with soluble nutrients, 48 hours.
Next I will try the normal amount of tea but enriched.

   


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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23831796 - 11/14/16 10:17 AM (7 years, 2 months ago)

So the King oyster was getting most of its energy and nutrition from the tea solubles.
The 50% less tea + nutrition is only half as strong as full tea power.

I have setup full power enriched for testing.

   

Peptides provide 2% energy overall.


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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23831797 - 11/14/16 10:17 AM (7 years, 2 months ago)

Here we have full power enriched, but different mycelium. Black poplar and Nameko.
Left, Black poplar, bacteria infected. Right, Nameko, yeast infected.

Both samples may be infected with mold.

   


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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23832127 - 11/14/16 12:43 PM (7 years, 2 months ago)

Results in 2 hours, the bacterial Black poplar is the same as the bacterial King oyster.
Bacterial liquefying and some sinking. Some myc debris growing on the right.

The yeast infected Nameko has no liquefying, no yeast activity.

Here is the bacterial Black poplar sample.

 

Both samples have fresh growth.


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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23832130 - 11/14/16 12:45 PM (7 years, 2 months ago)

The bacteria should stall in 24-48 hours like before.


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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23834916 - 11/15/16 09:47 AM (7 years, 2 months ago)

Enriching the T-Gel is a no no, it increases bacterial activity x10, and counters all inhibitory chemicals.
However, the yeast has stalled totally. The yeast infected Nameko has now only Nameko myc.

           

If you need more potency add more tea, use lime water to balance Ph if needed.
I will make a second attempt for the Black poplar on normal T-Gel.


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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23839094 - 11/16/16 03:02 PM (7 years, 2 months ago)

I'm looking forward to seeing the black poplar:thumbup:


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Re: Testing antibacterial starch free agars [Re: Marty Mycfly]
    #23840980 - 11/17/16 09:04 AM (7 years, 2 months ago)

I've decided to drop and replace my fertilizer for tea. More tea, more nutrients and antibacterial effects.
Tea contains both soluble and non soluble nutrients and energy. So now on only tea fertilizer.

The enriched T-Gel Nameko test, has failed with yeast and a small amount of bacteria.

Adding anything other than tea should be considered a backwards step.
Unless the materials used are antibacterial, and safe to use.

   

It is easy to spot contamination.


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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23840982 - 11/17/16 09:05 AM (7 years, 2 months ago)

So back to the original T-Gel recipe. I will do Black Poplar first.


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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23844783 - 11/18/16 10:08 AM (7 years, 2 months ago)

I made a batch of TGF-Tek today. After the microwave PC for 20 minutes, I took roughly 120g to use as spawn.
It then was assembled and pressure cooked, the rest I have placed in the freezer, ready to cook.

The substrate is perfect, not sticky, well hydrated and crumbly after the microwave.
I inoculated my sample with a King oyster peg from my first T-Gel test.

I'm doing 80% water content + 280g of water for cooking.


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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23848134 - 11/19/16 10:42 AM (7 years, 2 months ago)

Here is the King oyster peg onto TGF-Tek after 24 hours.
Recovered and re-growing, all nice and clean.

   


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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23850626 - 11/20/16 07:06 AM (7 years, 2 months ago)

Conclusively the mycelium brings up nutrients on dead hydrated wood.
Look no further than tea for a potent antibacterial fertilizer.

King oyster peg onto TGF-Tek after 48 hours.

   

Another peg onto plant food after 48 hours.

   


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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23857136 - 11/22/16 12:02 PM (7 years, 2 months ago)

I'm doing some further testing now on Ph, based on katbusa's findings. I am still waiting for my kit to arrive.
However whilst I wait, I assembled some T-Gel using lime water that I made, just to test it out.

The gelatin set faster and harder, no obvious difference in colour or smell.
I will use the same King oyster spawn used in my first T-Gel test.

Hopefully the lime does not neutralize any effects.


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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23859677 - 11/23/16 06:53 AM (7 years, 2 months ago)

Ok so after some research on the 4 active antibacterial ingredients in tea: Catechins, Tannins, Theaflavins, Theanine.
I can only go as far as Ph 6 before they become unstable. Thankfully oysters like a Ph range of 6 - 8.

With a tea based substrate a casing of Ph 7 - 8 can be used when fully colonized.

If are you looking for quick energy then destabilize tea.


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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23859762 - 11/23/16 07:53 AM (7 years, 2 months ago)

Here is the pre Ph kit sample I just assembled.

   

I made another to Ph test.


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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23862860 - 11/24/16 07:59 AM (7 years, 2 months ago)

Best educated guess I have ever made, managed to get Ph 6.2 - 6.4.

   

No active bacteria on the new T-Gel, KO regrowing in 24 hours.
Previous tests took 2-3 days for good regrowth.

   

No bacterial pooling or gelling.


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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23862912 - 11/24/16 08:19 AM (7 years, 2 months ago)

Mycelium almost always lowers pH to ~5 so when you raise it, it's just taking longer because growth is inhibited.


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Re: Testing antibacterial starch free agars [Re: drake89]
    #23862939 - 11/24/16 08:30 AM (7 years, 2 months ago)

I knew about the mycelium producing acids, I did not equate the reduction though, so good point.
On the lower Ph the bacteria stalled after 24 - 48 hours, so this will be interesting.

Will it stayed stalled, or grow a bit then stall again?


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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23863154 - 11/24/16 10:11 AM (7 years, 2 months ago)

Here is Ph 4 - 4.5 after 4 hours. Lots of bacterial activity.

   


Here is Ph 6.2 - 6.4 after 24 hours.

   


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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23863856 - 11/24/16 02:48 PM (7 years, 2 months ago)

This is the fastest and cleanest T-Gel yet, 30 hours:

   

No bacterial presence on the T-Gel.


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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23865530 - 11/25/16 09:26 AM (7 years, 2 months ago)

:threadmonitor:


Edited by 0yster (10/23/18 07:57 AM)


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Re: Testing antibacterial starch free agars [Re: 0yster]
    #23868540 - 11/26/16 10:46 AM (7 years, 2 months ago)

Here is day 3, at 20 - 22°C, fully dark in a cupboard.

   

Image quality due to condensate.


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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23868668 - 11/26/16 11:24 AM (7 years, 2 months ago)

Why do you need 2 accounts?


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Re: Testing antibacterial starch free agars [Re: spacechildo]
    #23868680 - 11/26/16 11:27 AM (7 years, 2 months ago)

Quote:

spacechildo said:
Why do you need 2 accounts?




I dunno but check out that ridiculous self rating.




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Edited by Inocuole (11/26/16 11:54 AM)


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Re: Testing antibacterial starch free agars [Re: Inocuole]
    #23869019 - 11/26/16 01:16 PM (7 years, 2 months ago)

Thats was before I got supporter for extra images, I forget to logout first sometimes  :blush:


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Re: Testing antibacterial starch free agars [Re: Ferather]
    #23869390 - 11/26/16 03:31 PM (7 years, 2 months ago)

Quote:

Ferather said:
Thats was before I got supporter for extra images, I forget to logout first sometimes  :blush:



But you rated yourself...


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