Home | Community | Message Board


High Mountain Compost
Please support our sponsors.

Mushrooms, Mycology and Psychedelics >> Advanced Mycology

Welcome to the Shroomery Message Board! You are experiencing a small sample of what the site has to offer. Please login or register to post messages and view our exclusive members-only content. You'll gain access to additional forums, file attachments, board customizations, encrypted private messages, and much more!

Shop:   Original Seeds Store Buy CBD, Cannabis Seeds, Compare CBD   Amazon Agar, Microscope, Paul Stamets, Petri Dish, pH Test Strips

Jump to first unread post. Pages: 1
InvisibleATWAR
Connoisseur

Registered: 01/26/03
Posts: 1,640
Loc: #108768 in line...
Agar: Isolation from molds...
    #2875170 - 07/10/04 03:58 AM (12 years, 10 months ago)

Ok, I have heard of adjusting the Ph of Agar to make it unsuitable to the growth of mold mycelium but still allow the growth of mushroom mycelium. I have also heard of sandwiching the contaminated mycelium or tissue between agar, allowing the mushroom mycelium to penetrate it but leave the mold behind. I have also noticed while browsing throug a catalog of mine a substance called Propionic acid and it says "mold inhibitor". I assume this would also inhibit the growth of mushroom mycelium as well, but I thought I would ask the experts here...

The reason I ask is I have recently just begun to clone wild edibles and find mold to be a problem from time to time. Is there any other techniques I can try, and what is a Ph I should aim for? Does the contam sandwich technique work well enough for me to try?


--------------------
To give is to live...



Post Extras: Print Post  Remind Me! Notify Moderator
OfflineMAIA
World-BridgerKartikeya (DftS)
Male User Gallery

Registered: 04/27/01
Posts: 7,275
Loc: Erra - 20 Tauri - M45 Sta...
Last seen: 1 year, 2 days
Re: Agar: Isolation from molds... [Re: ATWAR]
    #2885739 - 07/13/04 01:27 PM (12 years, 10 months ago)

Read this page about Propionic acid http://explanation-guide.info/meaning/Propionic-acid.html
Although it says it kills "some" bacteria and mycelium might survive, i'm also inclined to believe the substance could still be corrosive and damage the mycelium.
Look, i'm not an expert about this, if you want me to move this post to advanced cult. you'll probably have more answers. Or try making a new post at that forum.
Anyway, PM me about this if you want to.

MAIA


--------------------
Spiritual being, living a human experience ... The Shroomery Mandala



Use, do not abuse; neither abstinence nor excess ever renders man happy.
Voltaire


Post Extras: Print Post  Remind Me! Notify Moderator
InvisibleATWAR
Connoisseur

Registered: 01/26/03
Posts: 1,640
Loc: #108768 in line...
Re: Agar: Isolation from molds... [Re: MAIA]
    #2886073 - 07/13/04 02:38 PM (12 years, 10 months ago)

Quote:

MAIA said:
Although it says it kills "some" bacteria and mycelium might survive, i'm also inclined to believe the substance could still be corrosive and damage the mycelium.




This is what I figured as well. If it inhibits mold, it will probably inhibit mushroom mycelium as well. It doesn't hurt to ask though...

Quote:

Look, i'm not an expert about this, if you want me to move this post to advanced cult. you'll probably have more answers.




I was going to ask for that, but I was going to give it more time. Please, be my guest...


--------------------
To give is to live...



Post Extras: Print Post  Remind Me! Notify Moderator
OfflineRogerRabbitM
Bans for Pleasure
Male User Gallery

Registered: 03/26/03
Posts: 42,000
Loc: USA Mountain Northwest
Last seen: 1 month, 27 days
Trusted Cultivator
Re: Agar: Isolation from molds... [Re: ATWAR]
    #2888863 - 07/14/04 10:25 AM (12 years, 10 months ago)

A trick I've found with wild mushrooms, is to cut a small section out of the stem and put it between two pieces of moist cardboard. In a few days, mycelium will begin to grow. Take a strand of the new myc, and transfer it to antibiotic agar. The cardboard will have effectively 'filtered' most of the molds away. I take a bag of pre-cut 6" X 6" moistened cardboard when I go picking, for this purpose.

One trick to outrun bacteria(it isn't necessary for fungi imperfecti) is to make an agar sandwich as you mentioned. Place your culture on antibiotic agar, and place another disk of antibiotic agar on top of it. Take the very first myc to poke through the top layer, and transfer it to a new dish. Repeat the process. After a transfer or two this way, the bacterial rods should no longer be present. After you make the transfer, look at the old dish under the microscope. If the rods are gone, the specimen is clear.


Post Extras: Print Post  Remind Me! Notify Moderator
InvisibleATWAR
Connoisseur

Registered: 01/26/03
Posts: 1,640
Loc: #108768 in line...
Re: Agar: Isolation from molds... [Re: RogerRabbit]
    #2889325 - 07/14/04 01:38 PM (12 years, 10 months ago)

I have used cardboard before, it works. Although not with all species...


Quote:

RogerRabbit said:
One trick to outrun bacteria(it isn't necessary for fungi imperfecti) is to make an agar sandwich as you mentioned. Place your culture on antibiotic agar, and place another disk of antibiotic agar on top of it. Take the very first myc to poke through the top layer, and transfer it to a new dish. Repeat the process. After a transfer or two this way, the bacterial rods should no longer be present. After you make the transfer, look at the old dish under the microscope. If the rods are gone, the specimen is clear.




You mean something like this:
http://www.shroomery.org/forums/showflat...;o=&fpart=1


--------------------
To give is to live...



Post Extras: Print Post  Remind Me! Notify Moderator
OfflineWorkmanV
Psilocybe Microscopist
Male User Gallery

Registered: 03/01/01
Posts: 3,498
Loc: Washington, USA
Last seen: 2 days, 15 hours
Trusted Cultivator
Re: Agar: Isolation from molds... [Re: ATWAR]
    #2891580 - 07/15/04 12:31 AM (12 years, 10 months ago)

A small amount of benomyl (benlate) in your agar will inhibit molds but allow mushroom mycelium to grow. It is autoclavable so it can be added directly to your media before sterilization. I generally use a combination of 10mg benlate and 100mg gentamycin in 1 liter of agar media for badly contaminated spores or cultures. Putting much more than 10mg/liter of benlate in your agar will slow the mushroom mycelium to a crawl if it grows at all. It takes very little to stop molds.

Unfortunatley benlate was taken off the market a few years ago because of some issues with toxicity. But I think the main benomyl metabolite, carbendazim, is still available as a garden fungicide in the UK and Australia. Benlate (Dupont trade name) was the fungicide of choice for the button mushroom industry because of its selective effect on molds.

Benomyl (Benlate) is a non-water soluable powder that breaks down into carbendazim which is water soluable and is the active chemical that inhibits molds. I haven't used carbendazim directly myself.


--------------------
Research funded by the patrons of
The Spore Works
Exotic Spore Supply
Reinvesting 25% of Sales Towards Basic Research and Species Identification :amanitajar:


Post Extras: Print Post  Remind Me! Notify Moderator
InvisibleATWAR
Connoisseur

Registered: 01/26/03
Posts: 1,640
Loc: #108768 in line...
Re: Agar: Isolation from molds... [Re: Workman]
    #2891915 - 07/15/04 02:03 AM (12 years, 10 months ago)

Very interesting until I got to this part:
Quote:

Unfortunatley benlate was taken off the market a few years ago because of some issues with toxicity.




Unfortunate indeed. But you have given me something to search about...
This is the kind of thing we need in the archives...


--------------------
To give is to live...



Post Extras: Print Post  Remind Me! Notify Moderator
OfflineRogerRabbitM
Bans for Pleasure
Male User Gallery

Registered: 03/26/03
Posts: 42,000
Loc: USA Mountain Northwest
Last seen: 1 month, 27 days
Trusted Cultivator
Re: Agar: Isolation from molds... [Re: ATWAR]
    #2895090 - 07/15/04 11:55 PM (12 years, 10 months ago)

Atwar,
I do it similar to that. If I have bacteria riding along on the myc, I transfer a very small piece to a new petri dish of antibiotic agar. Then, take a second petri dish, and transfer the entire disk of agar to the top of the transfer you just made. You now have two full sized disks of antibiotic agar, with your contaminated myc in the middle. After a few days, myc will grow through the top layer and poke out, before reaching the edges of the dish. When myc pokes out the top, I use my inoculating loop to scrape some myc without taking any of the agar. Transfer this to a new dish. Somehow by growing right through the antibiotic agar, it seems to leave the bacteria behind.


Post Extras: Print Post  Remind Me! Notify Moderator
InvisibleATWAR
Connoisseur

Registered: 01/26/03
Posts: 1,640
Loc: #108768 in line...
Re: Agar: Isolation from molds... [Re: RogerRabbit]
    #2895681 - 07/16/04 02:20 AM (12 years, 10 months ago)

Quote:

RogerRabbit said:
Then, take a second petri dish, and transfer the entire disk of agar to the top of the transfer you just made




This is what I interpreted from Stamets in GGMM. I tried that but ran into problems holding on to the large piece. How do you transfer it from one dish to the other? I tried once but dropped it on the bench. So, I decided to use a small piece instead, it works just as well for me...

Quote:

Somehow by growing right through the antibiotic agar, it seems to leave the bacteria behind.




Yep... I wonder if it would work for mold isolation as well... It is worth a try, but I have no contaminated cultures anymore. Raising the Ph and isolating did the trick so far (along with placing the culture in the sun in one case). I have a fairly large experiment in the works that will encompass most of my time for the next month or so. But if I do clone anything else this year and run into mold, I will give this method a try (with slight modification). Plus I have a few other ideas as well that could prove useful, but I need to try them first...


--------------------
To give is to live...



Post Extras: Print Post  Remind Me! Notify Moderator
InvisiblePrisoner#1
Even Dumber ThanAdvertized!
 User Gallery

Registered: 01/22/03
Posts: 193,665
Loc: Pvt. Pubfag NutSuck
Trusted Cultivator
Re: Agar: Isolation from molds... [Re: ATWAR]
    #2898666 - 07/17/04 01:31 AM (12 years, 10 months ago)

Quote:

ATWAR said:


This is what I interpreted from Stamets in GGMM. I tried that but ran into problems holding on to the large piece. How do you transfer it from one dish to the other? I tried once but dropped it on the bench. So, I decided to use a small piece instead, it works just as well for me...







try loosening about 1/3 of the agar from the dish with your scalpel, surface tesnion will hold it long enough to invert and align the dishes anf the agar should drop right in...


Post Extras: Print Post  Remind Me! Notify Moderator
OfflineRogerRabbitM
Bans for Pleasure
Male User Gallery

Registered: 03/26/03
Posts: 42,000
Loc: USA Mountain Northwest
Last seen: 1 month, 27 days
Trusted Cultivator
Re: Agar: Isolation from molds... [Re: Prisoner#1]
    #2910879 - 07/21/04 01:49 AM (12 years, 10 months ago)

Yes, what Prisoner said is how I do it. Works like a charm.

I use the sandwich technique for isolation away from bacteria. For molds, simple antibiotic agar seems to do the trick.


Post Extras: Print Post  Remind Me! Notify Moderator
InvisibleATWAR
Connoisseur

Registered: 01/26/03
Posts: 1,640
Loc: #108768 in line...
Re: Agar: Isolation from molds... [Re: RogerRabbit]
    #2912886 - 07/21/04 05:55 PM (12 years, 10 months ago)

Quote:

RogerRabbit said:
I use the sandwich technique for isolation away from bacteria. For molds, simple antibiotic agar seems to do the trick.




I assume you got that backwards?
I had to ask...


--------------------
To give is to live...



Post Extras: Print Post  Remind Me! Notify Moderator
OfflineRogerRabbitM
Bans for Pleasure
Male User Gallery

Registered: 03/26/03
Posts: 42,000
Loc: USA Mountain Northwest
Last seen: 1 month, 27 days
Trusted Cultivator
Re: Agar: Isolation from molds... [Re: ATWAR]
    #2922229 - 07/23/04 11:37 PM (12 years, 10 months ago)

Nope...Didn't get it backwards. Normally, to get away from molds, a couple of transfers on antibiotic agar does the trick. Bacteria can be a whole different animal, as it seems to 'hitch hike' along on the mycelium. You can't see it without a scope, but it will sure contaminate your grain if you use it to inoculate your spawn. I'm talking the bacterial 'rods' and not something obvious that you can see like wet spot. I wish I had a camera attachment for my microscope.


Post Extras: Print Post  Remind Me! Notify Moderator
InvisibleATWAR
Connoisseur

Registered: 01/26/03
Posts: 1,640
Loc: #108768 in line...
Re: Agar: Isolation from molds... [Re: RogerRabbit]
    #2922330 - 07/24/04 12:27 AM (12 years, 10 months ago)

I fail to see why you would need antibiotic agar for the isolation from molds... IMO this is totally unnecessary, as it would have little to no effect on the mold whatsoever. Believe me, I know about the ability of bacteria to hitch a ride with the mycelium, which is why I decided to illustrate the sandwich tek for others to see. Bacteria is not the focus of this discussion, and is not nearly as difficult to isolate from IME when using antibiotics and the sandwich technique. Bacteria I have no problem with, but molds I do when cloning wild specimens...

That just sounds like wasteful and unnecessary use of antibiotics to me... Peroxide I can understand, but antibiotic agar?


--------------------
To give is to live...



Post Extras: Print Post  Remind Me! Notify Moderator
Jump to top. Pages: 1

Shop:   Original Seeds Store Buy CBD, Cannabis Seeds, Compare CBD   Amazon Agar, Microscope, Paul Stamets, Petri Dish, pH Test Strips

Mushrooms, Mycology and Psychedelics >> Advanced Mycology

Similar ThreadsPosterViewsRepliesLast post
* Agar isolations ?
( 1 2 all )
socratesmind 5,505 25 09/18/02 04:02 PM
by Anno
* Agar Isolation School Mobius_Strip 3,412 12 04/28/06 11:33 AM
by Mycomyth
* Is this a 'non-sectoring' agar isolate? Aeolus1369 6,355 11 12/01/09 12:24 AM
by MasFina
* Chill on the agar isolations for outdoor patch? b3jamboree 891 4 04/16/08 05:54 PM
by tahoe
* Creating a P. Cubensis with lower temperature parameters using agar ozzysmygod 1,197 13 05/06/09 12:23 AM
by 4hodmt
* Making a Good Substrain (Agar, etc.) JustAHobby 2,919 9 07/13/07 03:56 PM
by fastfred
* Creating an isolate
( 1 2 all )
No_Life_G33k 4,736 33 06/09/13 02:31 AM
by CaspuuuR
* Single Strain (spore) Isolation and the Benefits.. TrippinRhino 4,989 18 07/24/01 02:02 AM
by Anno

Extra information
You cannot start new topics / You cannot reply to topics
HTML is disabled / BBCode is enabled
Moderator: RogerRabbit
4,836 topic views. 0 members, 4 guests and 1 web crawlers are browsing this forum.
[ Toggle Favorite | Print Topic | Stats ]
Search this thread:
World Seed Supply
Please support our sponsors.

Copyright 1997-2017 Mind Media. Some rights reserved.

Generated in 0.042 seconds spending 0.006 seconds on 18 queries.