|
Some of these posts are very old and might contain outdated information. You may wish to search for newer posts instead.
|
Mycolorado
Hobbyist


Registered: 07/23/16
Posts: 8,538
Loc: Interdimensional Bootcamp
|
Cleaning dirty cultures with tea agar 9
#23860431 - 11/23/16 12:12 PM (7 years, 2 months ago) |
|
|
Just wanted to bring this to the attention of mush cult. katbusa and Ferather have been experimenting with tea agar to clean up contaminated spawn and cultures. I thought it would fit in over here as a great alternative to antibiotic agar...tea is readily available and not everyone wants to use antibiotics. Also, bacterial contams are often present when starting from prints and syringes, and can really be discouraging to those starting out. I modified the original recipe to use less tea as the original amount seemed a bit excessive. Here are a couple APE cultures and a malabar. All had some degree of bacterial contam and are now growing clear. Growth is very slow on this medium but will result in a clean culture. NOTE: spores will not germinate on this agar!

200ml H2O 4.25g black tea (2 bags) 2g ME 4g Agar

katbusa's tea agar thread https://www.shroomery.org/forums/showflat.php/Number/23771470
|
Josex
#cheat_code



Registered: 11/13/15
Posts: 8,995
Loc:
|
Re: Cleaning dirty cultures with tea agar [Re: Mycolorado]
#23860444 - 11/23/16 12:17 PM (7 years, 2 months ago) |
|
|
|
concretelush
shadow cast light mass



Registered: 08/22/16
Posts: 251
|
Re: Cleaning dirty cultures with tea agar [Re: Mycolorado]
#23860446 - 11/23/16 12:17 PM (7 years, 2 months ago) |
|
|
interesting might have to give this a whirl.
|
Ferather
Mycological



Registered: 03/19/15
Posts: 6,325
Loc: United Kingdom
Last seen: 1 year, 3 months
|
Re: Cleaning dirty cultures with tea agar [Re: concretelush] 1
#23860530 - 11/23/16 12:51 PM (7 years, 2 months ago) |
|
|
Thats almost the exact amount of Tea I use in my T-Gel, 6g per 250g water. Maximum stable Ph is 6, some lime balancing may be required.
Read more here and here. Thanks again Mycolorado.
|
Ferather
Mycological



Registered: 03/19/15
Posts: 6,325
Loc: United Kingdom
Last seen: 1 year, 3 months
|
Re: Cleaning dirty cultures with tea agar [Re: Ferather] 2
#23860533 - 11/23/16 12:52 PM (7 years, 2 months ago) |
|
|
T-Gel is experimental, however does not need pressure cooking, only a microwave.
|
Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,810
Loc: Canada
|
Re: Cleaning dirty cultures with tea agar [Re: Ferather] 3
#23860537 - 11/23/16 12:53 PM (7 years, 2 months ago) |
|
|
Nice work. I know of some who also use activated charcoal to help inhibit bacteria. Just goes to show that antibiotics are the last resort not the first.
|
Ferather
Mycological



Registered: 03/19/15
Posts: 6,325
Loc: United Kingdom
Last seen: 1 year, 3 months
|
Re: Cleaning dirty cultures with tea agar [Re: Pastywhyte] 3
#23860545 - 11/23/16 12:55 PM (7 years, 2 months ago) |
|
|
The most potent is cinnamon, according to experts it's the most potent natural antibacterial there is.
|
cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
Last seen: 2 years, 26 days
|
Re: Cleaning dirty cultures with tea agar [Re: Ferather]
#23860629 - 11/23/16 01:18 PM (7 years, 2 months ago) |
|
|
This is something I've seen before n the only drawback is exactly what you warned about. .spores do not germinate on it and this goes for mold spores as well...these you simply drag to to the next plate or to your jars where they will germinate. Nice write up
--------------------
  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
|
cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
Last seen: 2 years, 26 days
|
Re: Cleaning dirty cultures with tea agar [Re: cronicr]
#23860645 - 11/23/16 01:22 PM (7 years, 2 months ago) |
|
|
--------------------
  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
|
enlightenment
alchemist


Registered: 08/09/09
Posts: 1,647
Loc: Europe
Last seen: 9 months, 8 days
|
Re: Cleaning dirty cultures with tea agar [Re: cronicr]
#23860666 - 11/23/16 01:27 PM (7 years, 2 months ago) |
|
|
Quote:
spores do not germinate on it and this goes for mold spores as well...these you simply drag to to the next plate or to your jars where they will germinate.
Do you think the mold spores are spread by the mycelium growth? I never had this issue when I isolated healthy mycelium away from trichoderma.
If you transfer a piece of agar that is about 1,5 cm away from the "inoculation place" to another dish and make one more transfer for extra safety I doubt you have problems with trich in your jars.
|
cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
Last seen: 2 years, 26 days
|
Re: Cleaning dirty cultures with tea agar [Re: enlightenment]
#23860703 - 11/23/16 01:36 PM (7 years, 2 months ago) |
|
|
That's just it though....without trich germinating how do you know if it's there
--------------------
  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
|
Ferather
Mycological



Registered: 03/19/15
Posts: 6,325
Loc: United Kingdom
Last seen: 1 year, 3 months
|
Re: Cleaning dirty cultures with tea agar [Re: enlightenment] 2
#23860732 - 11/23/16 01:43 PM (7 years, 2 months ago) |
|
|
The benefit for spores not germinating is the huge reduction of accidental contamination. Both T-Gel and T-Agar can be assembled open air, not saying to do that.
I agree with enlightenment when it comes to Trich.
----
My first test sample of King oyster was infected with both bacillus cereus and trich. The bacillus stalled after 24-48 hours, only mycelium grew, even the trich.
I isolated several sections using sterilized toothpicks. Only the King oyster passed on to all the pegs.
|
Ferather
Mycological



Registered: 03/19/15
Posts: 6,325
Loc: United Kingdom
Last seen: 1 year, 3 months
|
Re: Cleaning dirty cultures with tea agar [Re: Ferather]
#23860739 - 11/23/16 01:46 PM (7 years, 2 months ago) |
|
|
Here is the same sample a few days previously.
|
cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
Last seen: 2 years, 26 days
|
Re: Cleaning dirty cultures with tea agar [Re: Ferather]
#23860742 - 11/23/16 01:47 PM (7 years, 2 months ago) |
|
|
I'm not saying there is anything wrong with it but just like anti biotic agar I would put through a standard recipe before going to grain spawn.
--------------------
  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
|
enlightenment
alchemist


Registered: 08/09/09
Posts: 1,647
Loc: Europe
Last seen: 9 months, 8 days
|
Re: Cleaning dirty cultures with tea agar [Re: cronicr]
#23860751 - 11/23/16 01:49 PM (7 years, 2 months ago) |
|
|
That would be a problem if the tea dish is used to inoculate. Maybe I don't get you right or don't understand what you mean, sorry.
After further transfers it eliminates the risk IMO.
Example what I am thinking about: - A tiny piece of an outdoor fruit (clone) to tea agar. - Mycelium grows, trich or bacteria mostly stays in the middle of the plate - Transfer a piece far away from the spot you placed the fruit sample to an agar dish without tea. - Wait for healthy growth. - Make another transfer - Use this to inoculate grain
I give it a try with a contaminated Mexicana A. syringe I did not toss yet. I keep you posted. I hope it works because this syringe has cost time and nerves. 
//edit
Quote:
cronicr said: I'm not saying there is anything wrong with it but just like anti biotic agar I would put through a standard recipe before going to grain spawn.
That's what I tried to say
|
Edmunter
Mr



Registered: 05/01/13
Posts: 5,699
Last seen: 1 month, 5 days
|
Re: Cleaning dirty cultures with tea agar [Re: cronicr]
#23860778 - 11/23/16 01:56 PM (7 years, 2 months ago) |
|
|
Can bacteria piggy back on mycellium from plate to plate. I really dont know what to do with these P.Sandose plates
Transfer 2 
Transfer 3 
Do you think this could help?
|
Ferather
Mycological



Registered: 03/19/15
Posts: 6,325
Loc: United Kingdom
Last seen: 1 year, 3 months
|
Re: Cleaning dirty cultures with tea agar [Re: enlightenment] 1
#23860786 - 11/23/16 01:58 PM (7 years, 2 months ago) |
|
|
Sorry you guys are right, I have not mentioned the order of process I mentioned in my main post.
Dirty sample > T-Agar or T-Gel > Isolate > Transfer, agar > Validate > Spawn.
|
cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
Last seen: 2 years, 26 days
|
Re: Cleaning dirty cultures with tea agar [Re: Edmunter]
#23860792 - 11/23/16 02:02 PM (7 years, 2 months ago) |
|
|
Quote:
Edmunter said: Can bacteria piggy back on mycellium from plate to plate. I really dont know what to do with these P.Sandose plates
Transfer 2 
Transfer 3 
Do you think this could help?
Yes this would help or try sandwich it
--------------------
  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
|
enlightenment
alchemist


Registered: 08/09/09
Posts: 1,647
Loc: Europe
Last seen: 9 months, 8 days
|
Re: Cleaning dirty cultures with tea agar [Re: Edmunter]
#23860794 - 11/23/16 02:02 PM (7 years, 2 months ago) |
|
|
Quote:
Edmunter said: Can bacteria piggy back on mycellium from plate to plate. I really dont know what to do with these P.Sandose plates
Transfer 2 
Transfer 3 
Do you think this could help?
You should give it a try. It is hard to get rid off bacteria. I gave up some dishes because of massive problems. Made up to 5 transfers. Bacteria is harder to defeat than trich because bacteria can grow very fast. It is hard to isolate healthy mycelium away. Antibiotic agar sounds like a godsend for this purpose.
Try it and enrich this thread with your experience.
|
Edmunter
Mr



Registered: 05/01/13
Posts: 5,699
Last seen: 1 month, 5 days
|
Re: Cleaning dirty cultures with tea agar [Re: cronicr]
#23860798 - 11/23/16 02:04 PM (7 years, 2 months ago) |
|
|
I will try this tek and post here, maybe along side a sandwich to compare
|
|