|
Zoda
Pilzkopf



Registered: 11/17/19
Posts: 39
Loc: Germany
Last seen: 2 days, 7 hours
|
Agar Dishes Infected or Not?
#26555614 - 03/24/20 05:58 PM (3 years, 10 months ago) |
|
|
Hello Shroomery,
after many failed attempts with liquid culture to grain, I have now prepared some Agar dishes and put single colonized grains onto them in my diy still air box.
The problem is, that most of them I guess are still contaminated with a bacteria that survives on the surface of the mycelium and is also white but looks more like a little cloud.
My question for the cutting in this specific case is, do I cut a pizza slice or just the crust so to speak? Since the outher rim seems to be able to flee from the bacteria,but they eventually always catch up ;(




Thank you for your help!
-------------------- “I’m growing mushrooms… because I can’t kill them. They just keep multiplying…and it’s like…I’m in service to them.”
|
Strainsfordaze


Registered: 05/10/18
Posts: 669
|
Re: Agar Dishes Infected or Not? [Re: Zoda]
#26555661 - 03/24/20 06:36 PM (3 years, 10 months ago) |
|
|
You can take an small slice anywhere that isn’t contaminated. Metabolic activity can be higher in the mycelial edge which is why some people tend to sample from there. Keep doing successive transfers away from the contaminations.
|
Sockadin



Registered: 01/03/10
Posts: 7,244
Last seen: 2 months, 20 days
|
|
Some of it looks good, what are you taking your plates with?
|
rumfor69
Bodhicitta Cultivator



Registered: 08/05/11
Posts: 6,767
Loc: In the Gills
Last seen: 44 minutes, 32 seconds
|
Re: Agar Dishes Infected or Not? [Re: Sockadin]
#26555786 - 03/24/20 07:45 PM (3 years, 10 months ago) |
|
|
That's some good looking mycelium in there. Time to do some transfers to new plates like suggested.
|
Zoda
Pilzkopf



Registered: 11/17/19
Posts: 39
Loc: Germany
Last seen: 2 days, 7 hours
|
Re: Agar Dishes Infected or Not? [Re: Sockadin]
#26556350 - 03/25/20 04:47 AM (3 years, 9 months ago) |
|
|
Quote:
Sockadin said: Some of it looks good, what are you taking your plates with?
Im not sure I understand the question?
I use
130ml Water 4g Malt Extract 2g Agar Powder
I pressure cook at 115 PSI for about 15-20 min, let it all cool down and then use my self made still air box. I will cut with a flame sterilized blade from the edges like suggested.
Thank you for the replies, I will do my first transfers today. My idea is that I take the 3 healthies looking plates and transfer two areas from them into a new petri dish.
-------------------- “I’m growing mushrooms… because I can’t kill them. They just keep multiplying…and it’s like…I’m in service to them.”
|
LogicaL Chaos
Ascension Energy & Alien UFOs




Registered: 05/12/07
Posts: 69,333
Loc: The Inexpressible...
Last seen: 1 hour, 37 minutes
|
Re: Agar Dishes Infected or Not? [Re: Zoda]
#26556467 - 03/25/20 07:29 AM (3 years, 9 months ago) |
|
|
It doesnt look infected to me They all look healthy from here.
|
jcm4620
Stranger


Registered: 05/26/19
Posts: 6,700
Last seen: 1 year, 11 months
|
|
he was prolly asking you what are you taping your plates with. lol to me it looks like moving box tape or something lol but hey use whats available i guess. your prolly better off just going and buying a roll of saran/cling wrap and cut it into 2 inch rolls. you can put in ziplock bags for extra protection if wanted also. it would just be easier to see and to prolly remove come transfer time. that sticky fuckin kinda tape is not your friend when wearing gloves lol👍
Edited by jcm4620 (03/25/20 08:33 AM)
|
Zoda
Pilzkopf



Registered: 11/17/19
Posts: 39
Loc: Germany
Last seen: 2 days, 7 hours
|
Re: Agar Dishes Infected or Not? [Re: jcm4620]
#26556627 - 03/25/20 08:58 AM (3 years, 9 months ago) |
|
|
Quote:
jcm4620 said: he was prolly asking you what are you taping your plates with. lol to me it looks like moving box tape or something lol but hey use whats available i guess. your prolly better off just going and buying a roll of saran/cling wrap and cut it into 2 inch rolls. you can put in ziplock bags for extra protection if wanted also. it would just be easier to see and to prolly remove come transfer time. that sticky fuckin kinda tape is not your friend when wearing gloves lol👍
haha yea it is moving tape so to speak, but i figured that also sterile and yes its a bitch with gloves on, but i somewhat managed to get all transfers going today:

And to the people wondering where the contamination is...its the stupid bacteria or whatever it is thats there that almost looks like mycelium just finer i would say...i tried to cut where i thought it was the "cleanest". one of the other dishes already has a reaction drop again:
-------------------- “I’m growing mushrooms… because I can’t kill them. They just keep multiplying…and it’s like…I’m in service to them.”
|
Zoda
Pilzkopf



Registered: 11/17/19
Posts: 39
Loc: Germany
Last seen: 2 days, 7 hours
|
Re: Agar Dishes Infected or Not? [Re: Zoda]
#26570382 - 04/01/20 04:39 AM (3 years, 9 months ago) |
|
|
Alright heres my update after 1 week:
The dishes look good, but Im starting to wondering if they just seem this way and the bacteria is still there.

Take a look at both samples from nr.2 in front of light they both look like mycelium, but the other has MUCH more white stuff on top of it. (/which part here is the bacteria?)


My specific questions are:
1. Is it possible that the bacteria is so interwoven with the mycelium (that it keeps growin on top of it), that I will never be able to seperate the two?
2. Are there other steps besides ordering another sterile sample that I can try to recover my strain?
3. How do I know that my sample is clean and uncontaminated if the bacteria basically looks like mycelium and also does not hinder its growth too much? It basically looks like healthy mycelium, but its contaminated (as multiple substrate bags that even fully colonized have shown)
-------------------- “I’m growing mushrooms… because I can’t kill them. They just keep multiplying…and it’s like…I’m in service to them.”
|
rumfor69
Bodhicitta Cultivator



Registered: 08/05/11
Posts: 6,767
Loc: In the Gills
Last seen: 44 minutes, 32 seconds
|
Re: Agar Dishes Infected or Not? [Re: Zoda]
#26570395 - 04/01/20 04:59 AM (3 years, 9 months ago) |
|
|
That looks like healthy mycelium in that most bottom pic.
I dunno about the interwoven not ever getting clean part but theres no such thing as sterile spores really, they grow in the open air and stuff gets on them.
If you had a fully colonized bag and it still had some bacteria in it you should still try to spawn it to bulk and try to get some fruit. You only need one fruit to grow so you can take a sample from the inside of the stem where the sample will be completely sterile. Do that in front of a flowhood and you're in like Flynn...or whatever I dunno who tf Flynn is or why he always gets in.
|
|