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Offlinekatbusa
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Antibacterial Tea Agar * 4
    #23771470 - 10/25/16 08:57 PM (7 years, 3 months ago)

I just recently completed cleaning a culture of Tarragon Oyster that ferather sent me. One of the methods I used to beat the bacterial infection that accompanied the mycelium was to make tea water agar plates. The reason behind this is because I do not like using antibiotics unless it is absolutley necessary. I guess I get this little tick from working in the wholesale plant business since a young age. There is nothing worse than having to deal with a pest that has a resistance to your pesticide or antibiotic. So I always look for natural, cleaner, safer methods before I go full nuclear.

So with that being said. Tea is not a one shot kills all solution. It is however affective against gram-positive and gram-negative bacteria but there are some bacteria that are resistant.

Here is a good research paper published on the web for our reading:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2763290/

Also check out ferathers Pocket Guide. He is doing some really good work with tea, carbon sources, starch free substrates,and more. He knows his shit and is worth a read!
https://www.shroomery.org/forums/showflat.php/Number/23725598/page/1

Here is my thread where I used tea agar to successfully clean a Tarragon oyster culture.
https://www.shroomery.org/forums/showflat.php/Number/23680797/page/1


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

So lets get onto the meat and potatos. The original post in this thread, is in essence going to be a living document. As I learn, muck up and break things I will post them and my thoughts. I also highly recommend people who are trying out tea to post what they observe and learn as well.

My original recipe is:

25 grams black tea
5 grams malt extract
9 grams agar
500ml water

Prepare it as you would for normal nutrient agar and PC for 25-30min. I used the sandwich method to clean the culture. The agar ends up being a nice dark brown color. Makes seeing mycelium a breeze yet you can still shine light through it.


The original recipe was intentionally heavy on the amount of tea used. I did this to find a known baseline for what worked. From here I plan on lowering the concentration till I find the bottom threshold where the effects of tea are minimal on bacteria. Then from there to find an optimal amount to be used.

Some things I noticed:

*At the 25gram per 500ml concentration the mycelium from a variety of different mushrooms expanded across the agar much slower than the normal half concentration malt extract agar that is used to clean mycelium. I believe this is due to the extra nutrients provided by the tea which I did not take into account.

*The agar seemed to be a bit softer than normal. I believe this could be due to a possible lower PH shift. I have had something similar happen when doing plant tissue culture. I need to verify this in future experiments.

*I have no idea if this is even tea related but I noticed that in two of my Oyster cultures the mycelium switched over to Rhizomorphic growth. I will keep an eye on future cultures to see if I can trend data to support this.

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

I had to delete one of my previous experiments due to being out of town and I wanted to reign in the preparation of the tea agar to be more consistent and controlled across the board. I will be using only pure green tea. No flavors or additives.

The Future preparation method will be

- Add desired amount of tea to 500ml simmering water. Let simmer for 5min
- Strain tea from the water and check water amount. Add more water if needed
    to bring back up to 500ml
- Check PH and adjust if desired or if needed for experiment
- Reheat water to simmering point
- Add 10 grams malt extract, stir to dissolve
- Add 9 grams agar, stir to dissolve
- Autoclave in pressure cooker for 25min
 
              **Done under Laminar Flow Hood**
- Let cool so agar container can be handle without burning ones self
- Gently stir solution to redistribute agar
- Add 20ml of tea agar solution to each  100mm plate
- Let agar plates cool and set to full strength under flow hood
- Use as needed

Each plate I will be using will have an X and Y axis written on the lid. The X will represent horizontal growth and the Y will represent the vertical growth. This will be used to measure the amount of growth across the surface of the agar.

I will also be keeping track of the temperature at the time of measurement and the speed of bacterial and mycelial growth over time. Plus any observations I might have.

The planned Tea concentrations are as follows:

25 grams per 500ml agar solution
20 grams per 500ml agar solution
15 grams per 500ml agar solution
10 grams per 500ml agar solution
5  grams per 500ml agar solution

If I get to the 5 grams concentration and there is still active antibacterial properties I will then step down 1 gram at a time till I see active bacterial growth. Then I will increase the concentration by .5 grams till I see antibacterial characteristics again. Hopefully this will be the threshold of tea and its antibacterial properties.

If at any point in time I notice the antibacterial properties stop before I reach the 5 gram concentration I will treat future experiments in the same way as if I reached the 5 gram concentration to find the threshold of tea's antibacterial properties.

Once the working threshold of tea is established the recipe for the TEK will be updated and future experiments pertaining to mycelium growth speed on tea agar will be conducted. Hopefully a happy medium will be established of good antibacterial properties that will not stunt the growth of mycelium.

I also eventually plan on putting all of the info from these experiments into a format that is easy to read.

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

25 gram per 500ml experiment:

Preparation:

- 25 grams of  green tea to 500ml simmering water. Let simmer for 5min
- Strain tea from the water and check water amount. Add more water if needed
    to bring back up to 500ml
- Did not check PH
- Reheat water to simmering point
- Add 10 grams malt extract, stir to dissolve
- Add 9 grams agar, stir to dissolve
- Autoclave in pressure cooker for 25min
 
              **Done under Laminar Flow Hood**
- Let cool so agar container can be handle without burning ones self
- Gently stir solution to redistribute agar
- Add 20ml of tea agar solution to each  100mm plate
- Let agar plates cool and set to full strength under flow hood

Samples:

-Two samples will be used.
  "Sample 1" -  A known active bacteria colony from a contaminated plate that
      is mycelium free. A smudge from the contamination will be made. The
      contaminated agar wedge will be placed next to the smudge.
 
  "Sample 2" - From known bacteria contaminated straw spawn that will have
      acitve mycelium growing along with the bacteria. 

-Four agar plates were prepared. Two plates of MEA nurtient agar with the 25 gram tea concentration and two plates of the MEA nutrient agar without tea.

11/27/2016:
     
    Sample 1 - Samples were established on tea and tea free MEA agar.
           
          Tea:
              - Measurments: X- 8.82mm  Y- 12.03mm
              - Temp: 70 Deg F

          No Tea:
              - Measurements: X- 8.10mm Y- 13.02mm
              - Temp: 70 Deg F

   
      Sample 2 -
       
          Tea:
              - Measurements: X- 22.72mm  Y- 17.70mm
              - Temp: 70 Deg F

          No Tea:
              - Measurements: X- 18.59mm Y- 19.54mm
              - Temp: 70 Deg F

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

11/30/2016:
     
    Sample 1
           
          Tea:
              - No reorganization or no new growth noticed with the bacteria
              - Measurments: X- 8.82mm  Y- 12.03mm
              - Temp: 71 Deg F
             

          No Tea:
              - Noticed the bacterial colony has reorganized and has also
                spread.
              - Measurements: X- 13.15mm Y- 19.02mm
              - Temp: 71 Deg F
             

   
      Sample 2 -
       
          Tea:
              - No new bacterial growth noticed. The straw has begun to
                slightly fuzz up from the mushroom mycelium
              - Measurements: X- 22.72mm  Y- 17.70mm
              - Temp: 71 Deg F
             
         
        No Tea:
              - The straw has begun to fuzz up from the mushroom mycelium. A
                very small string colony of bacteria has been noticed. Also
                a very small circular colony of bacteria has been noticed.
                However very hard to take a photo of.
              - Measurements:
                        String Colony: X- .47mm Y- 1.42mm
                        Circular Colony: X- 1.05mm Y- 1.03mm
              - Temp: 71 Deg F



     
         
     

   
 









Edited by katbusa (11/30/16 02:58 PM)


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OfflineQuadman
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Re: Antibacterial Tea Agar [Re: katbusa]
    #23771507 - 10/25/16 09:09 PM (7 years, 3 months ago)

Quote:



*At the 25gram per 500ml concentration the mycelium from a variety of different mushrooms expanded across the agar much slower than the normal half concentration malt extract agar. I believe this is due to the extra nutrients provided by the tea which I did not take into account.




I'm confused, did you mean to say faster?


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Offlinekatbusa
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Re: Antibacterial Tea Agar [Re: Quadman]
    #23771548 - 10/25/16 09:24 PM (7 years, 3 months ago)

No sir. The mycelium was slow to spread on the original formulation. When cleaning mycelium I use half or a little less than half of the called for nutrients. The reason for this is that the mycelium expands faster and has a chance to out run molds so you can get a small clean sample from the leading edge.

Now if you have a clean culture then I guess the speed at how it expands wouldn`t matter as much.


Edited by katbusa (10/25/16 09:25 PM)


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OfflineQuadman
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Re: Antibacterial Tea Agar [Re: katbusa]
    #23771634 - 10/25/16 10:05 PM (7 years, 3 months ago)

Ok , wanted to make sure I understood. Thanks


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OfflineFerather
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Re: Antibacterial Tea Agar [Re: katbusa]
    #23772284 - 10/26/16 04:06 AM (7 years, 3 months ago)

Very nice writeup, looking forward to more results and tests. Starting a few tests myself.
The extra nutrition part is true, 25g tea is like 60g starch free grain flour.


Quote:

katbusa said:
My one plate of Summer Oyster actually over grew the bacteria while in the fridge. Strong strain!



Summer oyster is more aggressive than Pink oyster, plus it generates a higher yield.


Edited by Ferather (10/26/16 06:42 AM)


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Offlineadadada
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Re: Antibacterial Tea Agar [Re: Ferather]
    #23772375 - 10/26/16 05:39 AM (7 years, 3 months ago)

What is this sandwich method that you speak of?  I searched through the forum and didn't find anything about it.

Quote:

I used the sandwich method to clean the culture.




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Offlinefilyep
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Re: Antibacterial Tea Agar [Re: adadada]
    #23772382 - 10/26/16 05:46 AM (7 years, 3 months ago)

I believe it's where you sandwich the transferred agar wedge between two antibacterial agar wedges. One being the receiving dish of agar and the other a wedge that you cut from another dish of antibacterial agar to cover the contaminated wedge. Hope that makes sense.

Or you do your transfer and make yourself a sandwich.  :smile:


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Offlinefilyep
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Re: Antibacterial Tea Agar [Re: filyep]
    #23772384 - 10/26/16 05:48 AM (7 years, 3 months ago)



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Re: Antibacterial Tea Agar [Re: filyep]
    #23772394 - 10/26/16 05:57 AM (7 years, 3 months ago)

Thank you!  Very interesting approach and a handy tool for cleaning cultures!

Now if only there was a similar method for separating out trich :smile:


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OfflineFerather
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Re: Antibacterial Tea Agar [Re: adadada]
    #23772429 - 10/26/16 06:44 AM (7 years, 3 months ago)

Quote:

adadada said:
Thank you!  Very interesting approach and a handy tool for cleaning cultures!

Now if only there was a similar method for separating out trich :smile:



Cant see why we cant try that after, just a case of energy source.
Something trich hates and your sample likes.


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

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OfflineFerather
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Re: Antibacterial Tea Agar [Re: Ferather]
    #23772860 - 10/26/16 10:12 AM (7 years, 3 months ago)

"Cinnamon verum bark has the highest antimicrobial activity, particularly against antibiotic resistant strains." -- Source.

New recipe: 20g of raw black tea, 1 teaspoon of ground cinnamon.



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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: Antibacterial Tea Agar [Re: filyep] * 1
    #23773521 - 10/26/16 02:39 PM (7 years, 3 months ago)

Quote:

katbusa said:
25 grams black tea
5 grams malt extract
9 grams agar
500ml water




Does it have to be black tea?

Quote:

filyep said:
Or you do your transfer and make yourself a sandwich.  :smile:



Can I use a SAB or do you need a flow hood for this? :wink:


--------------------
: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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Offlinekatbusa
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Re: Antibacterial Tea Agar [Re: flyontoast]
    #23773564 - 10/26/16 02:58 PM (7 years, 3 months ago)

No it does not have to be black tea. It can be either black or green. Green tea however seems to have better antibacterial properties. I chose black tea because my store was out of raw unflavored green tea.

Just an FYI even if you use green tea. It will end up darker much like normal tea after the PC process.

You do not need a Laminar flow hood for this. A SAB will work just fine. Just don't forget to add the pickle!! LOL


Edited by katbusa (10/26/16 03:02 PM)


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Offlinekatbusa
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Re: Antibacterial Tea Agar [Re: Ferather]
    #23773567 - 10/26/16 03:00 PM (7 years, 3 months ago)

Quote:

Ferather said:
Quote:

adadada said:
Thank you!  Very interesting approach and a handy tool for cleaning cultures!

Now if only there was a similar method for separating out trich :smile:



Cant see why we cant try that after, just a case of energy source.
Something trich hates and your sample likes.




Mold has always been a royal pain in the ass. I still have to rely on taking alot of samples till I end up with one or two that give cleanish mycelium to work with. I wish there was something we could do about slowing mold down.


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Offlinekatbusa
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Re: Antibacterial Tea Agar [Re: Ferather]
    #23773589 - 10/26/16 03:06 PM (7 years, 3 months ago)

Quote:

Ferather said:
"Cinnamon verum bark has the highest antimicrobial activity, particularly against antibiotic resistant strains." -- Source.

New recipe: 20g of raw black tea, 1 teaspoon of ground cinnamon.







Cinnamon 0_0. Holy smokes let us know how this works. I use cinnamon often after planting seeds in trays to prevent damping off disease which is a type of fungus. You might be on to something.. I wonder how trich and good mycelium will react to this.


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Re: Antibacterial Tea Agar [Re: katbusa]
    #23773612 - 10/26/16 03:12 PM (7 years, 3 months ago)






Mold has always been a royal pain in the ass. I still have to rely on taking alot of samples till I end up with one or two that give cleanish mycelium to work with. I wish there was something we could do about slowing mold down.



:whathesaid: Feather you need to reverse engineer something for us:grin:


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Re: Antibacterial Tea Agar [Re: Quadman]
    #23773628 - 10/26/16 03:15 PM (7 years, 3 months ago)

I never heard of that Katbusa , always used Benlate . Wonder how mycelium would react to that ?


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Offlinekatbusa
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Re: Antibacterial Tea Agar [Re: Quadman]
    #23773660 - 10/26/16 03:23 PM (7 years, 3 months ago)

Quote:

Quadman said:





Mold has always been a royal pain in the ass. I still have to rely on taking alot of samples till I end up with one or two that give cleanish mycelium to work with. I wish there was something we could do about slowing mold down.



:whathesaid: Feather you need to reverse engineer something for us:grin:




If anyone can do it, Ferather would be the one. His cinnamon idea is brilliant and makes sense. I hope it works


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Offlinekatbusa
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Re: Antibacterial Tea Agar [Re: katbusa]
    #23773670 - 10/26/16 03:28 PM (7 years, 3 months ago)

I know benlate / benomyl is systemic. Isn't is also broadspectrum for fungi?

I would imagine it would toast good fungi.


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Re: Antibacterial Tea Agar [Re: katbusa]
    #23773744 - 10/26/16 03:50 PM (7 years, 3 months ago)

As far as I see it cinnamon could be hit and miss like pine wood. Not all oyster's will eat pine, due to pine oils.
Cinnamon oil stops mold and bacteria for up to 60 days, so wee need to balance it, not too rich.

I don't mind slow as long as it works as intended, to clean samples.


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

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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Re: Antibacterial Tea Agar [Re: Ferather]
    #23774087 - 10/26/16 05:37 PM (7 years, 3 months ago)

Great work guys. Yes if we could find a way of slowing down the unfavourable fungus that would be awesome.


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Re: Antibacterial Tea Agar [Re: filyep]
    #23774138 - 10/26/16 05:51 PM (7 years, 3 months ago)

Benlate isn't organic, but perhaps to clean a culture an extremely low dose might work against mold.


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


Edited by Quadman (10/27/16 08:51 AM)


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Re: Antibacterial Tea Agar [Re: katbusa]
    #23775786 - 10/27/16 07:28 AM (7 years, 3 months ago)

So in terms of how antibacterial is tea, actually its a quadruple punch.

Catechins, Tannins, Theaflavins, Theanine.

Tea is perfect with grains.


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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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Re: Antibacterial Tea Agar [Re: Ferather]
    #23775950 - 10/27/16 08:52 AM (7 years, 3 months ago)

Further investigation suggests lower temperatures are then next logical step.
Even the slowest oyster mycelium can outgrow bacteria and mold.

Common molds are inhibited below 4°C, see here.
I can grow oysters at 2°C, although slow.

Tea agar > Fridge (2°C) > Isolate.


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

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

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

DTS DCH Driver for Realtek [DTS:X] - Unlocked and reprogrammed.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #23775952 - 10/27/16 08:54 AM (7 years, 3 months ago)

Further actions:

> Reduce total water content, 50% or less.
> Reduce O2 levels to minimum.
> Reduce condensate.


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

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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Edited by Ferather (10/27/16 09:48 AM)


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Offlinekatbusa
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Re: Antibacterial Tea Agar [Re: Ferather]
    #23780515 - 10/28/16 04:12 PM (7 years, 3 months ago)

You are a wealth of information Ferather! I get immense joy reading about what you post. Thank you!


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Re: Antibacterial Tea Agar [Re: katbusa]
    #23782335 - 10/29/16 10:56 AM (7 years, 3 months ago)

Another thing you could do, if you have a dropper such as an eye dropper.
Make a solution of apple vinegar and salt, and drop a bomb.

Selective destruction of trich effected areas.


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

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: Antibacterial Tea Agar [Re: Ferather]
    #23803525 - 11/05/16 02:20 PM (7 years, 2 months ago)

Has anybody tried doing tea as an LC? And do you think it could "clean" a contaminated LC?
I did a tea bag with 1.5 cups of water and a teaspoon of honey (assuming tea has nutrients) and made 3 LC jars. I cloned an oyster into 1 of the jars to see if it would work (not seeing much action yet, maybe I didn't put enough honey, but it's dark as hell). Also have a jar of contam oyster LC and was wondering if there was a way to clean it without an agar transfer... I know the answer is 'probably not', but here's to hoping for a silver bullet.


--------------------
:mushroom: My trade list :mushroom:
Looking for strong terrestrial fruiters for an outdoor beds experiment:
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Also, any commercially viable Pleurotus, cold or hot strains.
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Re: Antibacterial Tea Agar [Re: flyontoast]
    #23803834 - 11/05/16 04:16 PM (7 years, 2 months ago)

Tea inhibits bacterial growth, it doesn't really count as cleaning or sterilizing.
Hmmmm, try tea soaked card, then a few drops of LC in the middle.

The bacteria will likely be sugar or starch loving.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #23805737 - 11/06/16 08:46 AM (7 years, 2 months ago)

The tea agar I made is inhibiting pretty much all growth with the exception of blue oyster which is an animal.  King oyster and a couple cubes are just sitting on it and a variety of spores won't germinate on it.  I used Newman's Own and cut the tea content down.  Too rich or other thoughts?

200ml H2O
4.25g black tea (2 bags)
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Re: Antibacterial Tea Agar [Re: Mycolorado]
    #23805792 - 11/06/16 09:08 AM (7 years, 2 months ago)

My King oyster is also slow, with a black ring identical to yours.
I'm using only tea, water and gelatin, no sugar/starch.

Currently mine is attacking a trich contam.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #23805912 - 11/06/16 09:49 AM (7 years, 2 months ago)

That blue is aggressive, wish all others behave that way. My tea soaked dowels all colonized well except the king was slow . There must be something with the tea.


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Re: Antibacterial Tea Agar [Re: Quadman]
    #23805929 - 11/06/16 09:56 AM (7 years, 2 months ago)

Indeed very conclusive. And interesting, there's a particular chemical that is inhibiting it.
Ultimately it's not a big deal, since we will be transferring, just time consuming.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #23805967 - 11/06/16 10:09 AM (7 years, 2 months ago)

My guess would be that the tannins are responsible.  I'll make up some green tea plates to see how they do in comparison.


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Re: Antibacterial Tea Agar [Re: Mycolorado]
    #23805978 - 11/06/16 10:13 AM (7 years, 2 months ago)

Awesome thanks mate, every test helps. I might try adding some soluble fertilizer.


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Re: Antibacterial Tea Agar [Re: flyontoast]
    #23807743 - 11/06/16 06:43 PM (7 years, 2 months ago)

Quote:

flyontoast said:
Has anybody tried doing tea as an LC? And do you think it could "clean" a contaminated LC?
I did a tea bag with 1.5 cups of water and a teaspoon of honey (assuming tea has nutrients) and made 3 LC jars. I cloned an oyster into 1 of the jars to see if it would work (not seeing much action yet, maybe I didn't put enough honey, but it's dark as hell). Also have a jar of contam oyster LC and was wondering if there was a way to clean it without an agar transfer... I know the answer is 'probably not', but here's to hoping for a silver bullet.




Yes Im two weeks in on a tea water lc. I do not like how dark it is and will probably not use tea in LC again. I have several other tea based experiements running as well. I just havent had the time to do a full write up yet. Work has me tied down again. Working out of state leaves me limited time. Once Im back I plan on up dtating everything.

Also mycelium is slow on tea agar. One thing you should try is green tea. See if anything changes. Im also trying the green tea as well. Im starting to think that tea is going to be good for cleaning only. Or maybe the recipe is missing something.

Any thoughts?


Edited by katbusa (11/06/16 06:45 PM)


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Re: Antibacterial Tea Agar [Re: katbusa]
    #23807837 - 11/06/16 07:24 PM (7 years, 2 months ago)

Guys I think I might be on to something. While sitting on the throne typing up my previous comments I had an epiphany.

This whole time the signs were there with the agar but I didn't pay attention to them. I noticed my agar was a bit soft in my plates compared to my non-tea plates. Both have the same amount of agar.

Low PH causes agar to not set as hard as it should. I have experience with this doing tissue culture for plants. If the PH is off the agar sets too watery. We use alot more agar than whats called for in tissure culture but the principle is the same.

I have heard that a low ph can stunt the growth. There also have been a few posts here and there on shroomery and others stating that lower PH caused slower runs / colonization.

So I did a quick test. Tea is pretty acidic. This could be why the mycelium is slow to run on tea agar.

My tap was at 7.0ph with tea im getting a ph down to 5 to 4 depending on the tests. I did two tests. PH test strips and vial dropper type.

If any of you havebthe ability to test please do so to verify what I just did. I was using green tea btw.




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Re: Antibacterial Tea Agar [Re: katbusa]
    #23807930 - 11/06/16 08:02 PM (7 years, 2 months ago)

Low PH is an interesting variable. Would a pinch of lime adjust it, or would that further mess with the agar consistency? (I know nothing about agar, going to pour my first plates this week). I'm tempted to make a lime solution syringe and add it to my bluer oyster clone LC jar because the myc is really not doing anything. Might do that tomorrow if I've got the time.


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Re: Antibacterial Tea Agar [Re: flyontoast]
    #23807978 - 11/06/16 08:20 PM (7 years, 2 months ago)

Most mushrooms like an acidic environment and so does mold. Lime would slow your growth but it should still out compete mold. If I recall line is not recommend for agar though


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Re: Antibacterial Tea Agar [Re: vatman]
    #23808013 - 11/06/16 08:36 PM (7 years, 2 months ago)

I know mushrooms like acidic substrates. I'm just wondering if the PH from tea is too acidic for them.

I'm not trying to rewrite the books on what is considered common knowledge but there seems to be some conflicting info on the inter-web-of-things. Some people say to keep the substrate neutral. Some say low PH is fine. Some say low PH im stunted growth and so on.

I did how ever find this paper for oyster.

http://jabonline.in/admin/php/uploads/58_pdf.pdf

It says they saw optimal growth at a PH of 7 with the highest yeild. It also said this :

" However, the biological efficiency, biological yield and economic yield were increased
with the increase of pH levels up to 5.04 and then decreased"

Kind of conflicting.

I think I will try to adjust the PH to see what happens.



Edited by katbusa (11/06/16 08:50 PM)


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Re: Antibacterial Tea Agar [Re: katbusa]
    #23808022 - 11/06/16 08:40 PM (7 years, 2 months ago)

Looks more like a 5.5 from the photo but I could be wrong.


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Re: Antibacterial Tea Agar [Re: vatman]
    #23808054 - 11/06/16 08:52 PM (7 years, 2 months ago)

Web says black tea has a ph around 4.9.  Aloha meds say optimal sub ph is 5 - 6.5, for oyster anyway. From their site:

he optimal substrate pH value for mycelial growth is 5-6.5, though mycelium can survive between pH 4.2 and 7.5. The mycelium grows slowly as the pH lowers and stops growing at pH 4. If the pH is higher than the optimal value, mycelial growth accelerates but produces an abnormal structure. Optimal pH for primordial induction and fruiting is 5-5.5 though it is possible at 5.5-7.8. The pH of the substrate can be adjusted by the addition of gypsum or lime


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Re: Antibacterial Tea Agar [Re: Mycolorado]
    #23808125 - 11/06/16 09:20 PM (7 years, 2 months ago)

Quote:

vatman said:
Looks more like a 5.5 from the photo but I could be wrong.




The test strip got alot more yellow the longer it sat. The water was slightly red the first time then yellow the second time.

Quote:

Mycolorado said:
Web says black tea has a ph around 4.9.  Aloha meds say optimal sub ph is 5 - 6.5, for oyster anyway. From their site:

he optimal substrate pH value for mycelial growth is 5-6.5, though mycelium can survive between pH 4.2 and 7.5. The mycelium grows slowly as the pH lowers and stops growing at pH 4. If the pH is higher than the optimal value, mycelial growth accelerates but produces an abnormal structure. Optimal pH for primordial induction and fruiting is 5-5.5 though it is possible at 5.5-7.8. The pH of the substrate can be adjusted by the addition of gypsum or lime




I do not refute those PH numbers. I'm just more curious as to how the mycelium reacts to Tea at the low PH. The thing is that mycelium is slow on Tea agar. So every variable needs to be looked at. So I'm going to adjust the PH to 7 in my next agar batch to see if it speeds up. I honestly don't think it will make a difference. Gotta try though.


Edited by katbusa (11/06/16 09:42 PM)


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Re: Antibacterial Tea Agar [Re: katbusa]
    #23808163 - 11/06/16 09:39 PM (7 years, 2 months ago)

Digital ph readers would be the way to. Thought paper has a max time to be in water


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Re: Antibacterial Tea Agar [Re: katbusa]
    #23808164 - 11/06/16 09:40 PM (7 years, 2 months ago)

Fosho...just wanted to throw up some credible numbers.  What are you raising the ph with?


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Re: Antibacterial Tea Agar [Re: Mycolorado]
    #23808177 - 11/06/16 09:46 PM (7 years, 2 months ago)

Quote:

vatman said:
Digital ph readers would be the way to. Thought paper has a max time to be in water




I do have a digital reader but it isn't calibrated. Go figure on of the times I need it and I'm out of the packets to calibrate it.

Quote:

Mycolorado said:
Fosho...just wanted to throw up some credible numbers.  What are you raising the ph with?




I have no idea. I was honestly thinking of just using a pinch of baking soda. I've never had to adjust the PH of agar for mushrooms before so this is totally uncharted territory for me.


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Re: Antibacterial Tea Agar [Re: katbusa]
    #23808200 - 11/06/16 10:01 PM (7 years, 2 months ago)

Ha, that crossed my mind as well.  I was gonna try gyp but it says it won't neutralize acidity...lime will though.


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Re: Antibacterial Tea Agar [Re: Mycolorado]
    #23808300 - 11/06/16 11:17 PM (7 years, 2 months ago)

https://wfs.swst.org/index.php/wfs/article/view/149/149
Quote:

There was a tendency, although not perfect, for content of ethanol-soluble extractives to increase with increasing darkness of wood color. A distinct trend for water solubles in relation to wood color was not present. Table 1 also shows that as content of ethanol solubles increased, weight loss tended to decrease, or ill other words decay resistance appears to have been directly associated with the amount of ethanol-soluble extractives. The strength of this relationship is demonstrated by the results of the correlation analysis shown in Table 2, where the concentration of ethanol solubles accounts for 56 to 69% of the variation




i think this is talking about phobaphenes here, and i wonder if these alcohol soluble extractives can be useful to you guys in your endeavors somehow.


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Re: Antibacterial Tea Agar [Re: katbusa]
    #23808324 - 11/06/16 11:31 PM (7 years, 2 months ago)

Quote:

katbusa said:
Quote:

vatman said:
Digital ph readers would be the way to. Thought paper has a max time to be in water




I do have a digital reader but it isn't calibrated. Go figure on of the times I need it and I'm out of the packets to calibrate it.

Quote:

Mycolorado said:
Fosho...just wanted to throw up some credible numbers.  What are you raising the ph with?




I have no idea. I was honestly thinking of just using a pinch of baking soda. I've never had to adjust the PH of agar for mushrooms before so this is totally uncharted territory for me.




use soda ash. available at pool supply stores or in smaller amounts at aquarium stores as reef buffer.


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Re: Antibacterial Tea Agar [Re: filyep]
    #23808343 - 11/06/16 11:55 PM (7 years, 2 months ago)

Quote:

filyep said:
Great work guys. Yes if we could find a way of slowing down the unfavourable fungus that would be awesome.





I don't know if this will help or not, but according to these guys, garlic mustard has very strong antifungal effects on mycorrhizae.  of course, it might screw with all fungus for all i know.

http://journals.plos.org/plosbiology/article?id=10.1371/journal.pbio.0040140
Quote:

We then conducted additional experiments to confirm that garlic mustard specifically caused AMF decline in the native soils (Experiment 2–4). We grew seedlings of the same three native tree species used in Experiment 1 in uninvaded forest soils that were conditioned for 3 mo with either garlic mustard plants or with one of the three native tree species. All three tree species demonstrated significantly lower AMF colonization in soils conditioned by Al. petiolata (0%–10%) than in soils conditioned by the native plants (20%–65%; Figure 2A). AMF colonization was similar in unconditioned (control) soils and soils conditioned with native plants. In addition, growth of the tree seedlings was the lowest in soils conditioned by garlic mustard (Figure 2B), confirming that garlic mustard plants reduce native plant performance by interfering with the formation of mycorrhizal associations.






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Re: Antibacterial Tea Agar [Re: ballsalsa]
    #23808809 - 11/07/16 07:48 AM (7 years, 2 months ago)

Very nice read guys, I've already calculated the Ph based on data rather than testing.
Using my pocket guide, the average Ph range can be slightly different.

However the base would be 4.0-5.5, this guide, page 48.
Ph 3.5 reduced growth but not as much as Ph 4.5.

It's the chemicals in tea, not the Ph.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #23808834 - 11/07/16 08:05 AM (7 years, 2 months ago)

Found this on lime, apparently only tiny amounts in a fruiting substrate.
Then it becomes overdose, and inhibits all aspects of growth.

Edit: Wrong link to website.


Edited by Ferather (11/07/16 10:46 AM)


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Re: Antibacterial Tea Agar [Re: Ferather]
    #23809248 - 11/07/16 10:27 AM (7 years, 2 months ago)

Quote:

Ferather said:
Found this on lime, apparently only tiny amounts in a fruiting substrate.
Then it becomes overdose, and inhibits all aspects of growth.



you can disregard pretty much any study out of Africa and SE Asia...that's absolutely bogus.  Fungi mineralize rocks FFS.


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Re: Antibacterial Tea Agar [Re: drake89]
    #23809291 - 11/07/16 10:46 AM (7 years, 2 months ago)

Ok, thanks Drake. I also posted the wrong link, my bad. I ment this one for lime.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #23815652 - 11/09/16 09:16 AM (7 years, 2 months ago)

I take my comment back Mycolorado, my King oyster is strong, aggressive and normal speed for 20°C.
Something upsets your balance. Current difference: I am using gelatin, and no starch.

There is by far more energy in T-Gel, than T-Agar, Peptide's C-H-N-O.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #23816085 - 11/09/16 12:19 PM (7 years, 2 months ago)

Further notes:

The original clean Tarragon T-Agar sample sent by katbusa (Thanks again), lacked nutrition, but not energy or water.
The sample was left as assembled for 2 weeks with no action, I got growth in 8 hours on enriched tea.

I have already setup an enriched T-Gel test run, I am almost ready to transfer.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #23817529 - 11/09/16 08:12 PM (7 years, 2 months ago)

The king is starting to move and the blue ripped across it.  I have a gold oyster LC vendor syringe I'm gonna put to it as well as standard MEA to compare.  King and blue:


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Re: Antibacterial Tea Agar [Re: Mycolorado]
    #23818665 - 11/10/16 08:06 AM (7 years, 2 months ago)

Your King definitely suffers, mine is normal, its even strong enough to conquer trich.
Do you want me to send you a sample of this strong Gelatin I have?

You can side-by-side and investigate, no starch.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #23819538 - 11/10/16 11:58 AM (7 years, 2 months ago)

Thanks for the offer, Ferather, but I'll track some down!  This king has had some issues...it had a bacterial contamination when I got it and has never been what I would consider a strong specimen and could just be getting tired.  That said, I have some freshly germinated cube myc on the tea and it's not loving life either.


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Re: Antibacterial Tea Agar [Re: Mycolorado]
    #23819715 - 11/10/16 12:46 PM (7 years, 2 months ago)

Ok no probs. Here is my T-Gel recipe. It does not require starch, and no pressure cooker.
Make sure it's strong pig skin gelatin, other types will liquefy into a myco puddle.

Strong gelatin is much cheaper, 20g of gelatin = 10g agar, 14g starch.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #23820331 - 11/10/16 04:44 PM (7 years, 2 months ago)

Awesome info ferather. So you think the T-Agar I sent you was lacking in nutrients? When you say you go growth on enriched tea. Are you talking about your enriched tea slurry tek?

If not can you shed some light on what nutrients you added to enrich with?

I would not have even thought that there was a nutrient issue for the fact that the tea was adding nutrients. I'm writing this down now and I'm going to try adding more MEA to the agar, maybe even start with the original amount of MEA and just add tea water to see what happens.

Just got home from working out of town! Got my indoor greenhouse set up. The plan is to get some results posted this weekend and dial in my winter greenhouse setup as well. Going to be a good weekend!



Edited by katbusa (11/10/16 04:47 PM)


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Re: Antibacterial Tea Agar [Re: katbusa]
    #23820921 - 11/10/16 07:46 PM (7 years, 2 months ago)

The recipe I used was based on yours, katbusa, but I reduced the tea by over half. It was around 2% tea and 1% ME.  I was thinking along the same lines about MEA made with .5% or less tea.


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Re: Antibacterial Tea Agar [Re: Mycolorado]
    #23822279 - 11/11/16 08:29 AM (7 years, 2 months ago)

All I can say is the samples looked naked and dormant, the other sample still does.
8 hours after rolling and placing the sample it began to return to life.

The ratio is not high enough, or the PC destroys nutrients.

   

Enriched with fertilizer and grain flour.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #23823434 - 11/11/16 03:52 PM (7 years, 2 months ago)

Found a university study on mycelium and yeast, here. Page two I read about digestion and absorption.
Anyhow the answer is the solubility of the fertilizer. It can be instantly absorbed, and used.

The soluble fertilizer in this case counts as a nutritional accelerator.
Too much becomes toxic, like adding 20% sodium.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #23823898 - 11/11/16 07:01 PM (7 years, 2 months ago)

Not that anyone was eagerly awaiting this, but on the note of Ph:
I did make a syringe of lime water and injected it into my tea LC. It's been about 4 days and the blue oyster in there is still doing nothing. It either really doesn't like the amount of tea in there or I didn't put enough honey, or both.


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Re: Antibacterial Tea Agar [Re: flyontoast]
    #23825019 - 11/12/16 06:41 AM (7 years, 2 months ago)

Hmm, I have little use for LC, however I can do some side tests at some point.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #23865478 - 11/25/16 08:46 AM (7 years, 2 months ago)

Just throwing that in there for someone interested:
Oreganum vulgare essential oil  is highly antiseptic.

it contains Thymol and Carvacrol
https://en.wikipedia.org/wiki/Carvacrol

Quote:

Carvacrol inhibits the growth of several bacteria strains, e.g. Escherichia coli[6] and Bacillus cereus. Its low toxicity together with its pleasant taste and smell suggests its use as a food additive to prevent bacterial contamination.[7] In Pseudomonas aeruginosa it causes damages to the cell membrane of these bacteria and, unlike other terpenes, inhibits their proliferation.[8] The cause of the antimicrobial properties is believed to be disruption of the bacteria membrane.[9][10]




http://www.dailymail.co.uk/sciencetech/article-1088893/MRSA-wiped--OREGANO-say-British-scientists.html


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Re: Antibacterial Tea Agar [Re: Chk]
    #23865539 - 11/25/16 09:31 AM (7 years, 2 months ago)

Nice work, now I guess it needs to be tested on a fresh live mycelium sample.
Have you found any data in your research in regards to mycelium?

Another thing to look at is spore germination.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #23865720 - 11/25/16 10:51 AM (7 years, 2 months ago)

Didn't test it,

it's while reading the OP that i remembered it has antibacterial effect
i don't have any currently, could go buy some soon, there is an herbal shop close by.


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Re: Antibacterial Tea Agar [Re: Chk]
    #23865735 - 11/25/16 10:57 AM (7 years, 2 months ago)

Thats your choice either way, if you do get some I am interested in results.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #23872356 - 11/27/16 03:03 PM (7 years, 2 months ago)

I've put your samples on T-Gel (Ph6), one Summer and one Tarragon, they both responded and are growing.
Thank you so much again katbusa, Soon I should have both types growing on clean spawn.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #23872439 - 11/27/16 03:27 PM (7 years, 2 months ago)

http://www.nature.com/articles/srep19710

Is some more detailed evidence supporting ferather. I am still a little skeptical of the antibiotic properties lasting through the PC but green tea would be the way to go.


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Re: Antibacterial Tea Agar [Re: vatman]
    #23872556 - 11/27/16 03:57 PM (7 years, 2 months ago)



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Re: Antibacterial Tea Agar [Re: Ferather]
    #23872566 - 11/27/16 04:01 PM (7 years, 2 months ago)

Quote:

Ferather said:
Active ingrediants: Catechins (175 °C), Tannins (200 °C), Theaflavins (240°C), Theanine (174 °C). Very pressure cookable  :thumbup:



most of those compounds stated are more anti axodaints. Some are anti microbes. The anti microbes are more on the green tea side. So why did you start off black tea over green tea?


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Re: Antibacterial Tea Agar [Re: vatman]
    #23872572 - 11/27/16 04:03 PM (7 years, 2 months ago)

My reason was cheapness and they are easy to obtain, but by all means go for green tea, much clearer.


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Re: Antibacterial Tea Agar [Re: vatman]
    #23873253 - 11/27/16 08:01 PM (7 years, 2 months ago)

Quote:

vatman said:
Quote:

Ferather said:
Active ingrediants: Catechins (175 °C), Tannins (200 °C), Theaflavins (240°C), Theanine (174 °C). Very pressure cookable  :thumbup:



most of those compounds stated are more anti axodaints. Some are anti microbes. The anti microbes are more on the green tea side. So why did you start off black tea over green tea?




I'm sure you know this but green tea and black tea are from the same plant. Black tea is just fermented. Green tea is the stuff you want to use but black can do if green is not available.

Tea does work but there have been some side effects. For example some of my cultures seem to grow pretty slow on the stuff. While on of my blue oyst cultures ripped right through it in a matter of a week. I'm not sure the correlation. I think it might be a nutrient issue. Why would tea change this I have no idea.

It is without a shadow of a doubt anti-bacterial. In fact I have been having a hard time getting some of my tea plates to contaminate. Even trich is really slow on the stuff.

I have quite a few experiments going on right now with tea. I just wish my work would allow me to be home more so I can wrap them up. Winter is supposed to be slow but apparently not this winter. Storm after storm is causing power companies to require my time away.


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Re: Antibacterial Tea Agar [Re: katbusa]
    #23874261 - 11/28/16 06:14 AM (7 years, 2 months ago)

Keep it up katbusa, plenty of holes I missed in research. My best result was at Ph 6 via lime, close to normal in speed.
Bacteria was very aggressive @ Ph 4.5 - 5, Mycelium take acid damage over time, too in low Ph alone.

Blue and a few other types can tolerate Ph 4 - 9. King, Gold and others like Ph 6 - 8.

Bacterial activity at Ph 6 via lime is nil, void, zero.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #23874287 - 11/28/16 06:36 AM (7 years, 2 months ago)

Is that hydrated lime 4 adjusting.


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Re: Antibacterial Tea Agar [Re: Pinpapa]
    #23874844 - 11/28/16 10:52 AM (7 years, 2 months ago)

Quote:

Pinpapa said:
Is that hydrated lime 4 adjusting.



Dehydrated, powdered.

----

I am making an infected LC from dirty WBS spawn, both bacterial and yeast.
Using my own LC recipe, samples produced bubbles in 2 minutes.

When ready I will infect tea infused materials.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #23875037 - 11/28/16 12:04 PM (7 years, 2 months ago)

why lime? why not soda ash?


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Re: Antibacterial Tea Agar [Re: ballsalsa]
    #23875063 - 11/28/16 12:13 PM (7 years, 2 months ago)

Quote:

ballsalsa said:
why lime? why not soda ash?



Sodium, while trace is essential, too much will inhibit growth, gross overdose will kill mycelium.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #23877250 - 11/29/16 04:45 AM (7 years, 2 months ago)

Quote:

Ferather said:
Quote:

ballsalsa said:
why lime? why not soda ash?



Sodium, while trace is essential, too much will inhibit growth, gross overdose will kill mycelium.




Yes it without any doubt inhibit growth,
i tried doing PF-like cake with old bread (so it's around 15gr per Kg sodium chloride)
it worked but the colonization was damn slow, and the yield suffers.


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Re: Antibacterial Tea Agar [Re: Chk]
    #23877304 - 11/29/16 05:42 AM (7 years, 2 months ago)

I tried steamed bread once, in a PC. Breadmakers add salt as a preserve.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #23878505 - 11/29/16 02:41 PM (7 years, 2 months ago)

Turns out black tea is oxidized tea, not fermented, from source.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #23878607 - 11/29/16 03:09 PM (7 years, 2 months ago)

Some (not all) black tea are fermented with Aspergillus Niger

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


"A. niger is the main agent in the fermentation of Pu-erh tea,[10] the Okinawan awamori spirit, and some varieties of shochu"


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Re: Antibacterial Tea Agar [Re: Chk]
    #23878616 - 11/29/16 03:11 PM (7 years, 2 months ago)

help to identify please!


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Re: Antibacterial Tea Agar [Re: Dodgewelder87]
    #23878622 - 11/29/16 03:12 PM (7 years, 2 months ago)

Commonly translated as dark, or black tea (this type of tea is different from what in the West is known as: black tea, which in China is called: red tea).

Pu-erh tea would be labelled "Pu-erh tea" and not black tea.


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Re: Antibacterial Tea Agar [Re: katbusa]
    #23881982 - 11/30/16 03:04 PM (7 years, 1 month ago)

edited and updated


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Re: Antibacterial Tea Agar [Re: katbusa]
    #23882023 - 11/30/16 03:16 PM (7 years, 1 month ago)

Dam you beat me to it, so thats where you have been hiding. Very nice update!



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Re: Antibacterial Tea Agar [Re: Ferather]
    #23882081 - 11/30/16 03:31 PM (7 years, 1 month ago)

Thanks for the kind words ferather!


I had this experiment totally finished but a coworker used my camera and deleted my photos. So I had to start all over again. I noticed this sitting in the back of a helicopter looking at previous powerline photos. I could of strangled the SOB.


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Re: Antibacterial Tea Agar [Re: katbusa]
    #23885730 - 12/01/16 08:24 PM (7 years, 1 month ago)

That blows!


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Re: Antibacterial Tea Agar [Re: Mycolorado]
    #24454009 - 07/03/17 09:11 AM (6 years, 6 months ago)

I am trying another [experimental] recipe today, I've essentially eliminated sugars.
Here is the composition of tea, ready to drink, no additives (sugar, milk).

125g > CaCO3 water, boiling hot.
1 > Cheap black tea bag.
0.4g > 240B gelatin.
2g > Agar.

----

Stew the tea bag into the hot CaCO3 water using a jug, squeeze the bags on the sides for 5 minutes using a fork.
The idea is to extract as much from the tea bag as possible, increasing the carbon and nutrient value.

The CaCO3 water will help keep the pH in check as tea extracts are acidic by nature.
Add your tea extract and calcium water to your agar and gelatin powder.

----

Microwave the mixture for 40-80 seconds, if needed.


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Re: Antibacterial Tea Agar [Re: katbusa]
    #24458305 - 07/05/17 01:03 AM (6 years, 6 months ago)

:takingnotes:


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Re: Antibacterial Tea Agar [Re: Lennybernadino]
    #24458745 - 07/05/17 08:59 AM (6 years, 6 months ago)

Well the "T-Gel" agar is working, my sample peg also has cellulose bacteria. It's very similar in speed as normal agar.
No signs of the cellulose bacteria transferring yet, the wood peg will also provide carbon and nutrients.

This recipe is intended to clean dirty samples, spore's should not germinate.

       


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24458775 - 07/05/17 09:19 AM (6 years, 6 months ago)

The peg will also receive nutrients from all through the agar, depending on the height of the peg.
This is due to evaporation, capillary action and mycelial uptake (distribution).


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24459278 - 07/05/17 01:41 PM (6 years, 6 months ago)

You got to love "pro mycelial" or "carbon controlled" medium's, that have a "live mycelium preferred" label.

List of carbon sources, remaining required nutrients from the tea and gelatin.
The nutrients come in a natural and organic water soluble format.


Calcium bicarbonate: [ Ca | C | H | O3 ] 2.
Polyphenols, varied: [ C6 | H5 | O1 ] x.
Proteins, varied: [ Cx | Hx | Ox | Nx ] x.

Theophylline: C7 | H8 | N4 | O2.
Theobromine: C7 | H8 | N4 | O2.
Caffeine: C8 | H10 | N4 | O2.
Theaflavin: C29 | H24 | O12.
Tannins: C76 | H52 | O46.
Catechin: C15 | H14 | O6.
Carotene: C40 | H56.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24461360 - 07/06/17 08:07 AM (6 years, 6 months ago)

Strong, quick and fairly uniform growth, mycelium wins.

   


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24464543 - 07/07/17 09:19 AM (6 years, 6 months ago)

Sorry for today's image quality, I have condensate, the room hit 28°C, currently at 26°C.
Little regrowth on the peg, lots of mycelial activity on the T-Gel agar medium.

Tea turns black when oxidized, in this case due to laccase.
You should see a black ring around any radials.

   


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24464933 - 07/07/17 11:54 AM (6 years, 6 months ago)

Here are better more clear images for today, no signs of any contamination.
Slightly slower than normal malt extract agar, however still quick.

   


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24471633 - 07/10/17 02:30 AM (6 years, 6 months ago)

Can you just set and forget a foraged wild mushroom, say cordyceps into the T-gel without treating the mushroom body with iodine, iso, etc?

Edit : Sorry I misinterpreted your message on "sample" I thought it was mushroom fruitbody or something else but turns out you meant mycelium


Edited by DailyShroomer (07/10/17 02:34 AM)


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Re: Antibacterial Tea Agar [Re: DailyShroomer]
    #24471828 - 07/10/17 06:13 AM (6 years, 6 months ago)

Yes, any live sample, eventually the tissue will decay, but you should get transferable growth.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24471838 - 07/10/17 06:26 AM (6 years, 6 months ago)

Can you use substitute for CaCO3 like with powdered eggshell for example?? I totally want to try this out.


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Re: Antibacterial Tea Agar [Re: DailyShroomer]
    #24471860 - 07/10/17 06:43 AM (6 years, 6 months ago)

Yes, egg shell is 95-97% CaCO3, other names: Chalk, Limestone, Oyster shell, Egg shell, Agricultural lime.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24471981 - 07/10/17 07:43 AM (6 years, 6 months ago)

Sweet!!

By the way, have you seen this? https://www.shroomery.org/forums/showflat.php/Number/17356799

Is the T-agar essentially the same as the link I'm referencing or is that just some sort of upgraded PDA?? It will be more efficient to use pour agar tek I think if you have smaller PC I think.


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Re: Antibacterial Tea Agar [Re: DailyShroomer]
    #24471995 - 07/10/17 07:51 AM (6 years, 6 months ago)

Thanks for the link, no the T-Gel recipe is not based of that. The recipe is based on carbon content and enzymes.
However, I admit both the tea extract and CaCO3 are considered antibacterial, so yes there's a bonus.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24472044 - 07/10/17 08:22 AM (6 years, 6 months ago)

Can you use pour instead of no pour for T-gel like from my link?? I have a small electric PC, if can I'll buy loads of 45ml glass jars and start with them since it would be uber efficient to not PC the jars and just let the tea, CaCO3 and heat to do the work.


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Re: Antibacterial Tea Agar [Re: DailyShroomer]
    #24472060 - 07/10/17 08:32 AM (6 years, 6 months ago)

Yes, pour or no pour, entirely up to you. Handle as you would with any other agar.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24472083 - 07/10/17 08:50 AM (6 years, 6 months ago)

Sweet! :wink:


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Re: Antibacterial Tea Agar [Re: DailyShroomer]
    #24472085 - 07/10/17 08:52 AM (6 years, 6 months ago)

Oh, one more thing! What's the consensus on green tea?? Is it workable with the beef gelatin and the CaCO3??


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Re: Antibacterial Tea Agar [Re: DailyShroomer]
    #24472089 - 07/10/17 08:56 AM (6 years, 6 months ago)

Yes, a few other community members have tried green tea, and got positive results.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24480435 - 07/13/17 02:34 PM (6 years, 6 months ago)

I added my transfer pegs yesterday, it has already begun transferring.

       


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24486654 - 07/16/17 05:56 AM (6 years, 6 months ago)

The medium is more populated than I expected, as in there is not many gaps between the growth, a wide spread.
There is also signs of genetic differences, several areas are much faster and denser than others.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24486657 - 07/16/17 05:56 AM (6 years, 6 months ago)

In conclusion, my T-Gel agar recipe works as intended, without sugar, starch and cellulose contamination dropped to none.
I suggest assembling and working with it in a SAB, but open air is more than doable without a SAB-other.

Only organisms capable of re-assembling materials into essential sugars will grow.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24488421 - 07/16/17 09:33 PM (6 years, 6 months ago)

This T-Gel works best for dirty wild samples, right? Oh, btw about green tea. How much green tea powder for 125 gr of CaCO3 water?? Green tea usually comes in dissolvable powder and I don't think 12 gr per 125 gr water is a realistic ratio, that'll be too much tea.


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Re: Antibacterial Tea Agar [Re: DailyShroomer]
    #24489133 - 07/17/17 09:32 AM (6 years, 6 months ago)

Any dirty sample, wild or not, the recipe I am using uses 1 tea bag, which is 3.25g.
Given only the extract is used, I'd say that is up to 1g of extract, roughly.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24495218 - 07/19/17 10:34 PM (6 years, 6 months ago)

*noted


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Re: Antibacterial Tea Agar [Re: DailyShroomer]
    #24495723 - 07/20/17 05:53 AM (6 years, 6 months ago)

No problem, the recipe is cheap and easy to make, and works like other antibiotic agar's.
I suggest pH strips, you can add lime to the tea extract to pH 7-8 (oysters).

Then add the mixture to the dry agar and 240B gelatin.

:thumbinator:


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24496149 - 07/20/17 11:28 AM (6 years, 6 months ago)

I'm trying another recipe, using just CaCO3 as the primary carbon source.

125g > Boiling hot water + 10g CaCO3.

0.1g > MG soluble nutrients.
0.2g > 240B gelatin.
2.5g > Agar.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24496152 - 07/20/17 11:29 AM (6 years, 6 months ago)

I am transferring one of the pegs I produced using the T-Gel recipe.

   


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24500279 - 07/22/17 06:27 AM (6 years, 6 months ago)

The C-Gel agar is working, faster than T-Gel agar, and about the same speed as malt extract agar.
Both the MG solubles as nutrients and CaCO3 as carbon, are working as I intended.

Growth is slightly thicker than T-Gel, however still radial as normal.

   


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24500326 - 07/22/17 07:26 AM (6 years, 6 months ago)

The CaCO3 and MG nutrients are classed as inorganic, so essentially the C-Gel agar is inorganic.


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Edited by Ferather (07/22/17 07:39 AM)


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24506767 - 07/25/17 09:11 AM (6 years, 6 months ago)

The low carbon C-Gel agar works, and does not stall, I will be transferring using carbon rich pegs.
Use the C-Gel agar for extreme cleaning or validation of another transfer (2 step).

No contamination, discolouration or signs of any weirdness.

   


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24506776 - 07/25/17 09:19 AM (6 years, 6 months ago)

The next T-Gel agar recipe will be based on eatyualive's, 50% tea extract 50% water method.


Recipe: 62g CaCO3 water, 62g Extract, 2.5g Agar, 0.2g 240B Gelatin, 0.1g MG.

Extract: 3.25g (1 bag) Black tea + 125g CaCO3 water.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24510578 - 07/26/17 09:38 PM (6 years, 6 months ago)

Do you think your use of tea on agar will also have practical use in LC as well?? I'm interested in the tannin and polyphenol contents of tea as well as the acidic nature to deter growth of bacteria and mold in an LC jar, but perhaps for very narrow species of mushrooms.


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Re: Antibacterial Tea Agar [Re: DailyShroomer]
    #24510808 - 07/26/17 11:55 PM (6 years, 6 months ago)

Polysaccharide, polyphenol and tannic acid. With the first as food for your desired mycelium and the latter 2 as antibiotic to fight against contaminants. Don't you think this is how the T-gel works in a nutshell?? Also I think the C-gel might not work as good for acidic loving species e.g wood lovers.


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Re: Antibacterial Tea Agar [Re: DailyShroomer]
    #24511453 - 07/27/17 10:13 AM (6 years, 6 months ago)

T-Gel works in two ways, by restricting the carbon source (no sugar, starch or cellulose), it requires laccase and protease enzymes.
The second is the antibacterial effects of the phenol carbon sources (tannin, catechins), an inhibitory carbon source.

By default a wood lover will decay inhibitory materials in wood, the tree's natural defences.
Not all woodloving mycelium like acidity, in fact BE with oysters is higher @ 7-8.

I don't use liquid cultures, and therefore cannot comment on it.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24511479 - 07/27/17 10:33 AM (6 years, 6 months ago)

Have you seen this, Ferather?? https://www.shroomery.org/forums/showflat.php/Number/15736192/fpart/1/vc/1

Do you think they also have antibacterial properties like tea?? Do you think it will be better if amended with CaCO3?? I thought if that work so why tea wouldn't??

Also in my country there's a traditional wood tea drink made from Sappan Wood (relative to Brazil Wood) and they often sold in dried form so maybe I can do the same as link above with less wood to water ratio since they're dried?? What piqued my interest was the antimicrobial substance called Brazilin from these Sappan Wood (Biancaea sappan).
http://www.sciencedirect.com/science/article/pii/S1995764515000541

What do you think??


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Re: Antibacterial Tea Agar [Re: DailyShroomer]
    #24511603 - 07/27/17 11:50 AM (6 years, 6 months ago)

Wood extracts will work like tea extracts yes, just ignore that and call it "plant extract".
You want the soluble inhibitory (anti-microbial) materials, they are acidic.

The CaCO3 is used to achieve a desired end pH (not too acidic).
I've seen wood extract agar's, works of course.

Example plant products.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24511622 - 07/27/17 11:57 AM (6 years, 6 months ago)

Also to point out, the green machine is not only a mycelium, but also a woodlover.
Therefore other than higher pH, infection from spores is very likely.

So such thing as woodloving mediums without the green.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24511687 - 07/27/17 12:22 PM (6 years, 6 months ago)

Open air safe, non sterile, settings: 100% complex carbon source + pH 7-8.
No soluble or easy carbon sources + undesired pH (no activation).

You will still get "live" mold from spawn or transfers.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24511697 - 07/27/17 12:27 PM (6 years, 6 months ago)

You can also use my cellulose agar alternative, just control the end pH.
Paper pellets are 95-97% cellulose, no lignin, sugar etc.

25g pellets + (75g CaCO3 water + 1g MG).



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Re: Antibacterial Tea Agar [Re: Ferather]
    #24512485 - 07/27/17 06:32 PM (6 years, 6 months ago)

Just like I suspected, CaCO3 amendment for plant extract is a must, eh??


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24512492 - 07/27/17 06:34 PM (6 years, 6 months ago)

Btw, any alternative for MG???


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Re: Antibacterial Tea Agar [Re: DailyShroomer]
    #24513399 - 07/28/17 05:57 AM (6 years, 6 months ago)

look for Nigari, it's magnesiumchloride.


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Re: Antibacterial Tea Agar [Re: LizardWizard]
    #24513664 - 07/28/17 09:33 AM (6 years, 6 months ago)

No sodium or magnesium based pH adjustment, they have known toxicity levels, use calcium based alkali's only.
Any organic or non-organic "soluble nutrients", check the label, you want 1% nitrogen total added.

Gr0wer uses: Espoma organic plant-tone (5-3-3), Mycolorado uses: Age Old Grow (12-6-6).

I'm using Miracle-Gro (24-8-16), there are a few brands around.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24513688 - 07/28/17 09:50 AM (6 years, 6 months ago)

Here are the results of the updated T-Gel recipe, twice as strong, twice as fast.
Sorry for the image quality, there is condensate on the lid currently.

Scrapings also grow out, but growth is best from the peg.

   


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24513715 - 07/28/17 10:04 AM (6 years, 6 months ago)

Wait, hold up, you're trying to tell me Nigari has toxins in it or something? It's extracted out of sea salt, why would that be a problem? I don't see the point here, but maybe I'm not following... Please explain?


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24513735 - 07/28/17 10:17 AM (6 years, 6 months ago)

Btw is it okay to use pour tek without sterilizing plates for T-gel??


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Re: Antibacterial Tea Agar [Re: LizardWizard]
    #24513737 - 07/28/17 10:18 AM (6 years, 6 months ago)

Quote:

LizardWizard said:
Wait, hold up, you're trying to tell me Nigari has toxins in it or something? It's extracted out of sea salt, why would that be a problem? I don't see the point here, but maybe I'm not following... Please explain?



Sodium. Fungi hate that stuff.

Edit : Nevermind. I must be too tired, it was magnesium all along, I don't have much to say. Carry on.


Edited by DailyShroomer (07/28/17 10:21 AM)


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Re: Antibacterial Tea Agar [Re: DailyShroomer]
    #24513964 - 07/28/17 12:47 PM (6 years, 6 months ago)

It's not a problem in small amounts, but both sodium and magnesium cause issues at certain amounts-ratios.
Calcium is safe because it doesn't cause toxicity, only growth reactions to pH level are an issue.

However CaCO3 will stop dissolving above pH 9-10, so it's not actually that bad for pH.
Additionally no one sodium or magnesium bathes straw, because it doesn't work.

Furthermore its suggested to use low magnesium lime for this reason.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24513974 - 07/28/17 12:54 PM (6 years, 6 months ago)

Certain amounts-ratios of sugar and sodium are classed as "preservative", however at lower amounts both are "essential".


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24515017 - 07/28/17 09:18 PM (6 years, 6 months ago)

Do you think the usage of "plant extract" as nutrition for fungi also provide them with humic and fulvic acid which are readily available in nature??


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Re: Antibacterial Tea Agar [Re: DailyShroomer]
    #24515183 - 07/28/17 11:17 PM (6 years, 6 months ago)

Also what do you think about alkaline extraction of plant extracts from tea and wood instead of boiling them?? Just use your CaCO3 water without boiling.


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Re: Antibacterial Tea Agar [Re: DailyShroomer]
    #24515850 - 07/29/17 10:23 AM (6 years, 5 months ago)

CaCO3 does pull out more extractives, but I still use boiling hot water.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24516004 - 07/29/17 11:27 AM (6 years, 5 months ago)

Here are today's images of the peg and scraping on the T-Gel.
Need's a little more tea extract for a bit more carbon.

   


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24521621 - 07/31/17 09:39 PM (6 years, 5 months ago)

Btw can you use colonized toothpicks to inoculate grain jars?? Or are they only for transferring purposes??


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Re: Antibacterial Tea Agar [Re: DailyShroomer]
    #24522147 - 08/01/17 07:06 AM (6 years, 5 months ago)

If they have been sterilized, and you're using correct conditions (SAB, flowhood), do whatever you want.
Bearing in mind that the peg should be fully colonized, leaving little room for bacteria growth.

You may prefer to transfer the peg to an agar medium, and use that instead.
If you have clean agar, you can transfer many pegs, replace them.

:thumbup:


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24522708 - 08/01/17 12:29 PM (6 years, 5 months ago)

I updated my T-Gel antibacterial agar post, see my signature for the two recipe's.


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Re: Antibacterial Tea Agar [Re: DailyShroomer]
    #24523306 - 08/01/17 06:10 PM (6 years, 5 months ago)

Quote:

DailyShroomer said:
Btw can you use colonized toothpicks to inoculate grain jars?? Or are they only for transferring purposes??




If they are taken from wild clones, no. If taken from in vitro clones, maybe. If taken from mono or SGFC type grow, probably not. If taken to agar first to clean up, yes on all three. Grains are too prone to contamination to use colonized toothpicks unless they have been colonized in sterile conditions, so if they were colonized inside a sterile jar with a sterile inoculant with sterile technique, then yes. If it's a wood loving species, you wouldn't need to go to grains first and things would be easier.


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Re: Antibacterial Tea Agar [Re: LizardWizard]
    #24523609 - 08/01/17 08:37 PM (6 years, 5 months ago)

Thanks. I was thinking about agar and I think it will be a neat idea to do transfers like agar 2 agar. But I take that as a yes.


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Re: Antibacterial Tea Agar [Re: DailyShroomer]
    #24523612 - 08/01/17 08:38 PM (6 years, 5 months ago)

Ferather, if I were to use other plant extract other than tea, can you comment on the potency for germinating spores??


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Re: Antibacterial Tea Agar [Re: DailyShroomer]
    #24523690 - 08/01/17 09:23 PM (6 years, 5 months ago)

Transfers from agar to agar using wooden sticks have been around for a while, search for stareatclouds' staresticks TEK.


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Re: Antibacterial Tea Agar [Re: LizardWizard]
    #24524465 - 08/02/17 09:38 AM (6 years, 5 months ago)

I've seen wood extractive agar, however wood is not very macro-micro nutrient rich.
Other than tea, I have not tried much else. Malt extract composition, here.

You want near equal macro-micro nutrients, no sugar-starch.
Here is the composition of ready-to-drink tea.

----

The following replaces starch, sugar and cellulose (carbohydrates).

Calcium bicarbonate: [ Ca | C | H | O3 ] 2.
Proteins, varied: [ Cx | Hx | Ox | Nx ] x.
Polyphenols, varied: [ C6 | H5 | O ] x.

Theobromine: C7 | H8 | N4 | O2.
Theophylline: C7 | H8 | N4 | O2.
Caffeine: C8 | H10 | N4 | O2.
Theaflavin: C29 | H24 | O12.
Tannins: C76 | H52 | O46.
Catechin: C15 | H14 | O6.
Carotene: C40 | H56.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24524677 - 08/02/17 11:23 AM (6 years, 5 months ago)

Whoa, can you explain that in a bit of a layman term, I mean there are some success with wood broth liquid culture for wood lovers like P. cyanescens and all, is that due to starch sugar content in wood? So basically you're saying it is safer to use inorganic macro micro nutrients??


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Re: Antibacterial Tea Agar [Re: DailyShroomer]
    #24524933 - 08/02/17 01:27 PM (6 years, 5 months ago)

Starch and sugar can be utilized (digested) by almost any organism, including humans. Effectively you can germinate anything using them.
Wood loving mycelium germinate using the extractives (water solubles) in wood and other starch and sugar free plants (or nearly).

When making tea or wood extracts, you are extracting the soluble carbon that actually activates and germinates them.
However many of the common contamination you get (non-woodloving) do not utilize or activate using them.

Wood is weak in nitrogen and other macro-micro nutrients, tea leaves are rich in nutrients.
Therefore using wood extracts will give you plenty of carbon, but not much else.

It's not important if the added nutrients are organic or not.


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Re: Antibacterial Tea Agar [Re: Ferather]
    #24525904 - 08/02/17 08:37 PM (6 years, 5 months ago)

Quote:

Ferather said:
Open air safe, non sterile, settings: 100% complex carbon source + pH 7-8.
No soluble or easy carbon sources + undesired pH (no activation).

You will still get "live" mold from spawn or transfers.



Were you referring to the T-gel or C-gel??


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Re: Antibacterial Tea Agar [Re: DailyShroomer]
    #24526948 - 08/03/17 08:34 AM (6 years, 5 months ago)

That's a quick guide for reference to how open air or non-sterile methods work.
If you wanted to make or research a substrate, that's how they work.

Essentially it should have 0 or low germination chance.


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Re: Antibacterial Tea Agar [Re: Ferather] * 1
    #24552712 - 08/14/17 08:25 AM (6 years, 5 months ago)

Fantastic way to inhibit bacterial growth! I especially like the fact that tea is easy to come by and it's cheap.

Ferather could you (or someone else of course) write down the recipe in a way that even dummies (that would be me) can understand? I super like to try this with some bacterial mycelium I have :cool:. I eventually understood the recipe myself but I myself feel that many ppl. could benefit from a user friendly write-up that includes all info needed to prepare it; without the need to do additional research. I don't mean it in a bad way of course! Very good work. I appreciate all the hard work that has gone into it. It's some serious time investment you did there! Thanks a lot.

- What PH range should the TEA agar be, for most mushrooms?
- What do you mean with: "Any soluble macro-micro nutrient source" you mean malt extract or dextrose ect.?
- Is microwaving the tea agar enough? (because that is what it seems like in the recipe)
- What does the gelatin do?
- "Basic materials list, carbon based" (what do you exactly mean by this?)

The above are just some questions I love to have answered in the original recipe. For ease of use. I think it would help out a lot. You could also answer the above questions in a reply (maybe with some questions of others that might still pop up) and I will have a try rewriting your recipe. If necessary I will contribute... no problem...

thanks


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Re: Antibacterial Tea Agar [Re: Shroomway]
    #24555077 - 08/15/17 06:44 AM (6 years, 5 months ago)

I've essentially sat here for over 3 years doing nothing but research and alternative applications, I learned much from the community.
I have ADHD, which is both a gift and a burden, I can pick out things overlooked by others, including patterns or repeats.

So I apologize if my approach of sharing is "different" and perhaps "over informed".

----

1 - What PH range should the TEA agar be, for most mushrooms?
2 - What do you mean with: "Any soluble macro-micro nutrient source" you mean malt extract or dextrose ect.?
3 - Is microwaving the tea agar enough? (because that is what it seems like in the recipe)
4 - What does the gelatin do?
5 - "Basic materials list, carbon based" (what do you exactly mean by this?)

1 - This can change with the mycelium you are growing, which is why I did not state a pH range, 7-8 for oysters.
2 - Any source that contains no usable energy (carbon), you could call this A-Z vitamins and minerals.
3 - Yes, tea is very easy to make, add hot water. The other ingredients are already clean for use.
4 - Adds animal proteins (Gelatin is 97% protein), the proteins provide both carbon and nitrogen.
5 - This is the source of carbon (replaces the need for sugar), which is used as energy-other.


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Re: Antibacterial Tea Agar [Re: Ferather] * 1
    #24555514 - 08/15/17 10:53 AM (6 years, 5 months ago)

Ferather, thanks for the clarifications... Btw: we all have our strengths and pitfalls :smile:

I decided to keep it simple... I am cleaning myc with an tea agar. Experimenting with malt extract agar. Only using half the malt extract and tea in stead of water:
recipe for 500 ml's:
500 ml tea extract (made from 25 grams of green tea and water)
5 grams of malt extract
9 grams of agar

Will definately report back if I succeeded to clean out a really badly bacterialy infected petri...


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Re: Antibacterial Tea Agar [Re: Shroomway]
    #24556077 - 08/15/17 03:19 PM (6 years, 5 months ago)

Quote:

Shroomway said:
Ferather, thanks for the clarifications... Btw: we all have our strengths and pitfalls :smile:

I decided to keep it simple... I am cleaning myc with an tea agar. Experimenting with malt extract agar. Only using half the malt extract and tea in stead of water:
recipe for 500 ml's:
500 ml tea extract (made from 25 grams of green tea and water)
5 grams of malt extract
9 grams of agar

Will definately report back if I succeeded to clean out a really badly bacterialy infected petri...




:thumbup:


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