|
 Some of these posts are very old and might contain outdated information. You may wish to search for newer posts instead.
|
 BlueDisa
Friend
Registered: 03/02/17
Posts: 170
Last seen: 11 days, 22 hours
|
 Bacteria inhibiting spore germination 
#24329582 - 05/18/17 10:20 AM (6 years, 10 months ago) |
|
|
Good Day Shroomers
I have some prints of different species that I am trying to germinate.
Some of the prints are contaminated with bacteria.
When I put spores on agar to germinate, the bacteria germinates first and consequently my spores do not germinate (as the bacteria grows all over the spores) so I do not even have a chance to isolate some healthy mycelium. How do I get these infected spore prints to germinate and produce some mycelium? I have read about antibiotic agar, but I live rural and won't be able to get any of the mycologically useful antibiotics any time soon.
I have got some Wine Caps that managed to germinate right through the bacterial contamination, I did an agar sandwich with them and got some clean mycelium.
Are there any teks I could follow to help with this problem.
|
 bodhisatta  
Smurf real estate agent
Registered: 04/30/13
Posts: 61,891
Loc: Milky way
|
Re: Bacteria inhibiting spore germination [Re: BlueDisa]
#24329596 - 05/18/17 10:24 AM (6 years, 10 months ago) |
|
|
How are you putting the spores on the agar?
I suggest if from a spore print to take a loop to remove some spores from the print then take a small amount of sterilized water in a shot glass and dip the loop in. This dilutes the concentration or spores down and also bacteria. Then dip a new sterile loop into the water/spore solution then make streak plates.
Streaking out spores rather than blobbing a drop in the middle of a plate will greatly increase your chance of success
|
36fuckin5
Alchemycologist
Registered: 08/11/03
Posts: 12,086
Loc: Diving into Mystical Territori...
|
Re: Bacteria inhibiting spore germination [Re: bodhisatta]
#24329712 - 05/18/17 11:04 AM (6 years, 10 months ago) |
|
|
Quote:
bodhisatta said:
How are you putting the spores on the agar?
I suggest if from a spore print to take a loop to remove some spores from the print then take a small amount of sterilized water in a shot glass and dip the loop in. This dilutes the concentration or spores down and also bacteria. Then dip a new sterile loop into the water/spore solution then make streak plates.
Streaking out spores rather than blobbing a drop in the middle of a plate will greatly increase your chance of success
Exactly what I was gonna suggest. With clean prints I don't bother with this, but if it's an outdoor print or something just not grown in the cleanest place then streaking is the way to go.
--------------------
Redd Foxx said:
If you're offended I don't give a shit and don't come see me no more.
Pat The Bunny said:
A punk rock song won't ever change the world, but I can tell you about a couple that changed me.
bodhisatta said:
i recommend common sense and figuring it out.
These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.
|
BlueDisa
Friend
Registered: 03/02/17
Posts: 170
Last seen: 11 days, 22 hours
|
Re: Bacteria inhibiting spore germination [Re: 36fuckin5]
#24331716 - 05/19/17 12:37 AM (6 years, 10 months ago) |
|
|
I have just been scraping spores off prints with a scalpel blade and placed it all in one spot on the dish , so that is probably my mistake.
Thank you for the advice, I will streak some plates and update with results. 
|
Alan Rockefeller
Mycologist
Registered: 03/10/07
Posts: 48,364
Last seen: 8 hours, 38 minutes
|
Re: Bacteria inhibiting spore germination [Re: BlueDisa]
#24332040 - 05/19/17 04:22 AM (6 years, 10 months ago) |
|
|
You can mail order antibiotics for your agar, here are the ones I use:
http://www.the-odin.com/ampicillin-sodium/
http://www.the-odin.com/kanamycin-monosulfate/
I did a test yesterday where I tried adding them before and after pressure cooking, and then put 3 species of bacteria on the dishes. Will know in a couple days if you can autoclave them or if you need to add them afterwards. I suspect that you can autoclave them.
|
gillaputty
Stranger
Registered: 05/11/17
Posts: 266
Loc: Earth
Last seen: 6 years, 10 months
|
|
Use dasani to reintake the minerAL intake of the gullet of the Mushroom veils.
Edited by gillaputty (05/19/17 03:50 PM)
|
bodhisatta
Smurf real estate agent
Registered: 04/30/13
Posts: 61,891
Loc: Milky way
|
Re: Bacteria inhibiting spore germination [Re: gillaputty] 6
#24333208 - 05/19/17 02:32 PM (6 years, 10 months ago) |
|
|
Quote:
gillaputty said:
Try excluding potatoes flakes in your agar.
Use dasani to reintake the minerAL intake of the gullet of the Mushroom veils.
|
c10h12n2o
serial dilutor
Registered: 01/21/15
Posts: 3,200
Loc: the abyss
|
Re: Bacteria inhibiting spore germination [Re: bodhisatta]
#24333757 - 05/19/17 06:20 PM (6 years, 10 months ago) |
|
|
thanks Alan! let us know how that turns out, I've never worked with those antibiotics
I have only used gentamicin and chloramphenicol, and only VERY rarely, since well-timed transfers and streaking techniques usually make it pretty simple to separate from bacteria
I've only come across two prints that were so dirty they required antibiotics (due to the bacteria suppressing it). also, sometimes I use them as a precaution on the final plate before making master slants
--------------------
  
C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide "Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing." "Convictions are more dangerous enemies of truth than lies" ― Friedrich Nietzsche
|
gillaputty
Stranger
Registered: 05/11/17
Posts: 266
Loc: Earth
Last seen: 6 years, 10 months
|
Re: Bacteria inhibiting spore germination [Re: c10h12n2o]
#24333773 - 05/19/17 06:26 PM (6 years, 10 months ago) |
|
|
How about it mixed.
Try 5 ml solution.
It exists to he six ml currently in my make-up.
Change the colingal passage way up and reuptake nakephederyn.
Edited by gillaputty (05/19/17 06:27 PM)
|
gillaputty
Stranger
Registered: 05/11/17
Posts: 266
Loc: Earth
Last seen: 6 years, 10 months
|
Re: Bacteria inhibiting spore germination [Re: gillaputty]
#24334142 - 05/19/17 08:36 PM (6 years, 10 months ago) |
|
|
|
36fuckin5
Alchemycologist
Registered: 08/11/03
Posts: 12,086
Loc: Diving into Mystical Territori...
|
Re: Bacteria inhibiting spore germination [Re: gillaputty]
#24339675 - 05/22/17 03:56 AM (6 years, 10 months ago) |
|
|
Quote:
gillaputty said:
Use dasani to reintake the minerAL intake of the gullet of the Mushroom veils.
Are these even words? WTF are you talking about?
--------------------
Redd Foxx said:
If you're offended I don't give a shit and don't come see me no more.
Pat The Bunny said:
A punk rock song won't ever change the world, but I can tell you about a couple that changed me.
bodhisatta said:
i recommend common sense and figuring it out.
These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.
|
Lennybernadino
Amazon grower
Registered: 09/16/09
Posts: 770
Loc: Iquitos, Peru
Last seen: 2 months, 12 days
|
|
Any luck with autoclaving your two trial antibiotic media?
|
Alan Rockefeller
Mycologist
Registered: 03/10/07
Posts: 48,364
Last seen: 8 hours, 38 minutes
|
Re: Bacteria inhibiting spore germination [Re: Lennybernadino]
#24386992 - 06/08/17 01:30 AM (6 years, 10 months ago) |
|
|
Quote:
Lennybernadino said:
Any luck with autoclaving your two trial antibiotic media?
The antibiotics still worked after being autoclaved.
I was using Ampicillin and Kanamycin. I am going to start adding chlorampheticol too - bacteria that are resistant to the ampicillin type antibiotics are pretty common in nature, far fewer are resistant to chlorampheticol, and you don't need to add very much.
|
c10h12n2o
serial dilutor
Registered: 01/21/15
Posts: 3,200
Loc: the abyss
|
|
great info as always, thanks alan!
i use gent + chloramph in combination when i use antibiotics (which isnt very often), really handy for knocking out hitchhiking bacteria or on the last plate before making slants
--------------------
C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide "Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing." "Convictions are more dangerous enemies of truth than lies" ― Friedrich Nietzsche
|
Chwyn
Bacteria Rancher
Registered: 08/21/16
Posts: 1,238
Last seen: 1 year, 1 month
|
Re: Bacteria inhibiting spore germination [Re: c10h12n2o]
#24387750 - 06/08/17 10:12 AM (6 years, 10 months ago) |
|
|
A user named ferather showed me this the other day. this is an old thread but id like to add.
Antibacterial agar: https://www.shroomery.org/forums/showflat.php/Number/23771470
although semi-experimental it does work.
It also works with gelatin + agar + lime + ME mixes.
|
|
Threaded
Previous
Index
Next
Edit 
Reply 
afoaf
Yachaj
trade_om
comario2
1 2 3 all )
MycoFactory
