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OfflineKreskin
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Possible Start Mycelium From Contaminated LC?
    #27806660 - 06/05/22 07:36 AM (1 year, 7 months ago)

I bought an LC syringe from a vendor that I have tried on grain, and 5 plates over 3 sessions. At first I thought it’s probably me but I diid clones and spores on agar each time in the same SAB as the LC and only the LC would fail with green mold showing up in a couple of days.
    Before I waste anymore resources, is there a way to separate the myc from the green mold? The mold is so aggressive that I don’t think it’s possible but there is so much experience on here that maybe someone has a method for this.
  Oh, the vendor claims they guarantee their products but they don’t reply to their support form lol! Thanks for any suggestions!


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OfflineDERRAYLD
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Re: Possible Start Mycelium From Contaminated LC? [Re: Kreskin]
    #27807805 - 06/06/22 02:27 AM (1 year, 7 months ago)

Lc to agar then transfer clean sectors to agar until you have a clean culture.


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OfflineKreskin
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Re: Possible Start Mycelium From Contaminated LC? [Re: DERRAYLD]
    #27807996 - 06/06/22 07:33 AM (1 year, 7 months ago)

The green mold is faster than the mycelium growth so I haven’t been able to do it.


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InvisibleCreonAntigone
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Re: Possible Start Mycelium From Contaminated LC? [Re: Kreskin]
    #27813538 - 06/10/22 03:35 PM (1 year, 7 months ago)

Quote:

Kreskin said:
The green mold is faster than the mycelium growth so I haven’t been able to do it.





Well obviously a person could tell you to start over and it'd be the best solution - but I don't think I need to tell you that. You would likely save weeks, maybe months, working with new spores over trying to save this project.

But if you're determined the save the culture or if it's your only one, maybe you are particularly motivated.

Unfortunately, what I'm seeing is that transfer is not possible right now because the green mold is indistinguishable from the myc. In essence it is acting as a parasite to it. You can see the mold is strongest where the myc is, so no transfer of a particularly promising section is possible.

I would suggest you try several transfers to antimicrobial media. Try out as many recipes as possible. Molds are more simple than mushrooms, so it is possible in a media that is specially formulated, the mushroom will be favored and the mold will die out.

First off try good old fashioned antibiotic agar. It's possible that the anti-bacterial will also be harmful to the molds, or that there is a hidden bacteria in culture that is favoring the mold. Also worth considering are supplementing a small amount of a broad spectrum anti-fungal in your medium - theoretically enough to slow things down but not kill everything, and thus the mushroom which has a more complex lifecycle will win.

If you lack access to professional pharmacological antimicrobials, you could try the following recipes:

-media with .5%, 1% and 5% tannins added (can be derived easily from soaking acorns or sawdust)
-media with antimicrobial herbal extracts, most promising would be neem, which has proven to cure fungal diseases in culture* - I would try media supplemented with 1-20 drops of neem oil, then a media with a more significant amount of neem.
-media with a slightly higher content of sugars, which is unfavorable to some contaminants. You could try a media made from honey with barely any dilution - I tried a transfer to a mostly-honey media from a contaminated culture, and the contaminant did not show itself when the honey content reached a certain level - honey is both too high in sugar for most microbes, and contains antimicrobials itself. A media with 'too much honey' might still enable a mushroom to survive, but eliminate the simpler molds and bacteria.

There are many more recipes for antimicrobial agar, some much better than these ones. The basic principle is to try to create a media where the mushroom will be favored over this green mold.


*I realize this is a strong claim that needs defense - here is the evidence. I am referring to this study which exposed Aspergillus cultures to neem volatiles. It found a 51% reduction in fungal biomass - so the fungi grew slower but was not killed - and, miraculously, cultures exposed to neem produced far less of the aflaxtoxin - a reduction of aflatoxin values of 90%!

Therefore exposure to neem might change a culture so that it becomes less harmful. I think neem-supplemented agar would help here, as it'd slow down the faster mold and give the mushroom culture a chance to recover. I'd try 5 drops of neem oil into the media recipe.


Edited by CreonAntigone (06/10/22 11:06 PM)


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OfflineKreskin
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Re: Possible Start Mycelium From Contaminated LC? [Re: CreonAntigone]
    #27814704 - 06/11/22 02:36 PM (1 year, 7 months ago)

Quote:

CreonAntigone said:
Quote:

Kreskin said:
The green mold is faster than the mycelium growth so I haven’t been able to do it.





Well obviously a person could tell you to start over and it'd be the best solution - but I don't think I need to tell you that. You would likely save weeks, maybe months, working with new spores over trying to save this project.

But if you're determined the save the culture or if it's your only one, maybe you are particularly motivated.

Unfortunately, what I'm seeing is that transfer is not possible right now because the green mold is indistinguishable from the myc. In essence it is acting as a parasite to it. You can see the mold is strongest where the myc is, so no transfer of a particularly promising section is possible.

I would suggest you try several transfers to antimicrobial media. Try out as many recipes as possible. Molds are more simple than mushrooms, so it is possible in a media that is specially formulated, the mushroom will be favored and the mold will die out.

First off try good old fashioned antibiotic agar. It's possible that the anti-bacterial will also be harmful to the molds, or that there is a hidden bacteria in culture that is favoring the mold. Also worth considering are supplementing a small amount of a broad spectrum anti-fungal in your medium - theoretically enough to slow things down but not kill everything, and thus the mushroom which has a more complex lifecycle will win.

If you lack access to professional pharmacological antimicrobials, you could try the following recipes:

-media with .5%, 1% and 5% tannins added (can be derived easily from soaking acorns or sawdust)
-media with antimicrobial herbal extracts, most promising would be neem, which has proven to cure fungal diseases in culture* - I would try media supplemented with 1-20 drops of neem oil, then a media with a more significant amount of neem.
-media with a slightly higher content of sugars, which is unfavorable to some contaminants. You could try a media made from honey with barely any dilution - I tried a transfer to a mostly-honey media from a contaminated culture, and the contaminant did not show itself when the honey content reached a certain level - honey is both too high in sugar for most microbes, and contains antimicrobials itself. A media with 'too much honey' might still enable a mushroom to survive, but eliminate the simpler molds and bacteria.

There are many more recipes for antimicrobial agar, some much better than these ones. The basic principle is to try to create a media where the mushroom will be favored over this green mold.


*I realize this is a strong claim that needs defense - here is the evidence. I am referring to this study which exposed Aspergillus cultures to neem volatiles. It found a 51% reduction in fungal biomass - so the fungi grew slower but was not killed - and, miraculously, cultures exposed to neem produced far less of the aflaxtoxin - a reduction of aflatoxin values of 90%!

Therefore exposure to neem might change a culture so that it becomes less harmful. I think neem-supplemented agar would help here, as it'd slow down the faster mold and give the mushroom culture a chance to recover. I'd try 5 drops of neem oil into the media recipe.





Thank you for the detailed reply. I will sure try some of those solutions. I wanted the variety that was in the LC but if nothing else, I will learn more about LC work and also to avoid a certain vendor.


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