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WATER AGAR


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#1 EYMAIOS

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Posted 06 January 2021 - 03:34 PM

We all know how hesitant the spores are in the beginning with nutrient agar.
An alternative approach according to some mycologists is the water agar.

Water Agar is recommended for enumeration, cultivation and observation of sporulation of fungi.

Composition:
Ingredients Gms Agar 15 -20 / Litre water

Directions
Suspend 15-20 grams in 1000 ml distilled water.
Leave it for a while to soak.

Heat to boiling to dissolve the medium completely.

Sterilize by autoclaving
at 15 lbs pressure (121°C) for 10 to 15 minutes.

 

 

If anyone has any experience on water agar please enlighten us...

 

https://www.youtube....h?v=mXIVhY-ODXs
 



#2 HrVanker

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Posted 06 January 2021 - 07:20 PM

Nutrition is extremely important for germinating spores. Once they germinate and start building hyphae, they need to start taking in nutes in order to continue the process and create a thick mat of myc. On top of that, there are specific triggers for the germination of spores. The major one is the presence of usable nutrition. Some mushrooms, like Psilocybe mutans for instance, are very fond of horse and cow dung and are resistant to germinating on anything else:

"McKnight found that 95% of the spores germinated in horse dung decoction, 94% in cow dung, 73% in pig dung, 28% in dog dung, and 26% in chicken dung deooctions. The decoctions were aqueous extracts of the dried dung. McKnight found that only 0.1 to 4.1% of the spores germinated in distilled water. Negative results were also obtained with 0.1% and 0.01% water solutions of malt extract, yeast extract, casein hydrolysate, and ribonucleic acid. Extracts of orange juice, of grass, and of alfalfa also gave negative results. Water solutions of furfural and hemoglobin resulted in 8.9% and 12.1% germination respectively. Spores treated in a water bath with temperatures ranging from 30°-60° C for periods of from 5 minutes to 5 hours did not germinate better than the water controls. However, 9.1% of the spores germinated when they were heated to 50° C for 15 minutes and then frozen for 24 hours." - Neal K. Van Alfen

 

Granted, that's only one species and cubensis seems to be a lot less picky when it comes to nutrition. However I suspect that if we want to find a miracle germination recipe, we have to look at the environment in which a species grows. In their natural environment cubes like cow and horse dung, so a manure agar like the one above would probably be our best bet for germinating difficult spores. Likewise, stubborn woodlover spores would probably get a germination boost from an agar made from wood broth.

 

Now, if you have a culture with some difficult bacteria, perhaps you could clean it up using water agar. In theory, the myc from the transfer could utilize the bit of nutrition from the transfer wedge to grow out a bit on the water agar, and the bacteria should starve.


Edited by HrVanker, 06 January 2021 - 07:22 PM.

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#3 EYMAIOS

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Posted 07 January 2021 - 02:35 AM

 

Nutrition is extremely important for germinating spores. Once they germinate and start building hyphae, they need to start taking in nutes in order to continue the process and create a thick mat of myc. On top of that, there are specific triggers for the germination of spores. The major one is the presence of usable nutrition. Some mushrooms, like Psilocybe mutans for instance, are very fond of horse and cow dung and are resistant to germinating on anything else:

"McKnight found that 95% of the spores germinated in horse dung decoction, 94% in cow dung, 73% in pig dung, 28% in dog dung, and 26% in chicken dung deooctions. The decoctions were aqueous extracts of the dried dung. McKnight found that only 0.1 to 4.1% of the spores germinated in distilled water. Negative results were also obtained with 0.1% and 0.01% water solutions of malt extract, yeast extract, casein hydrolysate, and ribonucleic acid. Extracts of orange juice, of grass, and of alfalfa also gave negative results. Water solutions of furfural and hemoglobin resulted in 8.9% and 12.1% germination respectively. Spores treated in a water bath with temperatures ranging from 30°-60° C for periods of from 5 minutes to 5 hours did not germinate better than the water controls. However, 9.1% of the spores germinated when they were heated to 50° C for 15 minutes and then frozen for 24 hours." - Neal K. Van Alfen

 

Granted, that's only one species and cubensis seems to be a lot less picky when it comes to nutrition. However I suspect that if we want to find a miracle germination recipe, we have to look at the environment in which a species grows. In their natural environment cubes like cow and horse dung, so a manure agar like the one above would probably be our best bet for germinating difficult spores. Likewise, stubborn woodlover spores would probably get a germination boost from an agar made from wood broth.

 

Now, if you have a culture with some difficult bacteria, perhaps you could clean it up using water agar. In theory, the myc from the transfer could utilize the bit of nutrition from the transfer wedge to grow out a bit on the water agar, and the bacteria should starve.

 

Most illuminating!
Thank you indeed...
Question is what can we do to speed up the lazy nutrient agar process in relation to cow dung?



#4 HrVanker

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Posted 07 January 2021 - 03:01 AM

 

Question is what can we do to speed up the lazy nutrient agar process in relation to cow dung?

 

 

I'm not entirely sure what you mean. What is the "nutrient agar process" you speak of, and what is its relation to cow dung?



#5 jrh

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Posted 07 January 2021 - 07:20 AM

Now, if you have a culture with some difficult bacteria, perhaps you could clean it up using water agar. In theory, the myc from the transfer could utilize the bit of nutrition from the transfer wedge to grow out a bit on the water agar, and the bacteria should starve.

 

That's brilliant.


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#6 HrVanker

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Posted 07 January 2021 - 04:06 PM

 

Now, if you have a culture with some difficult bacteria, perhaps you could clean it up using water agar. In theory, the myc from the transfer could utilize the bit of nutrition from the transfer wedge to grow out a bit on the water agar, and the bacteria should starve.

 

That's brilliant.

 

I can't take too much credit for the idea. After writing the caveat about nutes, I did a little googling for water agar and came across a thread from Shroomery where it was mentioned.



#7 Jrotten

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Posted 07 January 2021 - 08:53 PM

Diluted water is the beat thing I’ve found for agar and LC.




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