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InvisibleMushouse
Mycomancer

Registered: 06/26/06
Posts: 500
Apple juice liquid culture (w/ Pics)
    #11047142 - 09/12/09 05:49 PM (7 years, 6 months ago)

I want to share a few pictures of cubensis liquid cultures using apple juice:

--How they got this far:
Sporeprints on aluminum foil were stored in the refrigerator for three years. A scraping of spores inoculated a petri dish of Potato Dextrose Agar (PDA). The petri dish became heavily contaminated with bacteria early, before any sign of mycelium could be seen. The contamination was due either to no sterile technique when the sporeprint was taken, or due to being stored in the fridge for three years (although the prints were sealed in plastic zip-lock bags).

After two or three weeks, two young patches of mycelium could be seen growing on the plate, although it had been contaminated by white bacteria from almost the beginning, and had just recently been contaminated by an unknown orange-yellow contamination with some sort of cob-webby sporulating hairs. A sample from each of the two mycelium patches was taken, and two new agar plates were made, each treated with a fine mist of hydrogen peroxide because they had come from such a heavily contaminated plate. The original, contaminated plate was sprayed with a mist of hydrogen peroxide and was kept for observation/curiosity (and the mycelium was later seen to do well on the contaminated plate, even growing over contamination).

The two new plates never contaminated, but took a long time to show any growth, due to the hydrogen peroxide, but eventually one of the two plates took off running and started spreading across the PDA. Some of the more promising growth was selected, and four transfers were taken to make four more petri dishes.

After some time, two of the four new petri dishes were showing rhizomorphic growth, and two liquid cultures were made from each of those two dishes.

--The liquid cultures:
Four baby food jars were taken and boiled in water along with eight glass beads that would later be used to agitate the liquid culture. Each lid had a small hole drilled in the top, and each hole was sealed with silicone to act as a self-healing port for a syringe needle. Each jar got two glass beads, and was filled with a bit of hydrogen peroxide. With the finished lids on, the jars were shaken for a while to sterilize them.

Each jar was drained of its hydrogen peroxide and each jar received a transfer of rhizomorphic mycelium. The jars were filled with apple juice and the lids were screwed on tight. With the assistance of the glass beads, the jars were shaken daily.

These pictures were taken exactly two weeks after the liquid culture jars were made:

Jar 1 was in the lead:



Jar 2 started showing growth a couple of days before this picture. This is what jar 1 looked like at first; many little points of growth suspended in the apple juice.


Jar 3 (no pic) was just a day or two behind jar 2. Jar 4 (no pic) had not yet shown any growth.


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Invisible13shroomsM
Lightning Shaman
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Registered: 01/01/09
Posts: 25,395
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Re: Apple juice liquid culture (w/ Pics) [Re: Mushouse]
    #11047213 - 09/12/09 06:06 PM (7 years, 6 months ago)

can u give more info on the apple juice?  was it 5% or 100% actual fruit juice?  you didnt pressure cook the juice or anything 4 that matter?


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Offlinenexus1946
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Re: Apple juice liquid culture (w/ Pics) [Re: Mushouse]
    #11047236 - 09/12/09 06:11 PM (7 years, 6 months ago)

Interesting. Nice work!

Straight apple juice eh? Straight from the can/jar?


--------------------
Gypsum/Drywall Tek
The Story Of Russell The Texas (Cube) Bear



The human race's prospects of survival were considerably better when we were defenceless against tigers than they are today when we have become defenceless against ourselves.
-Arnold J. Toynbee


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InvisibleMushouse
Mycomancer

Registered: 06/26/06
Posts: 500
Re: Apple juice liquid culture (w/ Pics) [Re: 13shrooms] * 1
    #11047295 - 09/12/09 06:25 PM (7 years, 6 months ago)

Quote:

13shrooms said:
can u give more info on the apple juice?  was it 5% or 100% actual fruit juice?  you didnt pressure cook the juice or anything 4 that matter?




It was a 6-pack of Minute Maid little 10oz apple juice bottles. The package said "100% Juice."

Ingredients: sure filtered water, concentrated apple juice, less than 0.5% of: vitamin C (ascorbic acid), calcium citrate, potassium phosphate

The generic grocery store brand 6-pack would have been used, because they are identical juice, but the Minute Maid bottles were bought for their shape, as the original plan was using them right in their own bottles.

The apple juice was not pressure cooked or treated in any way. Used straight out of the bottle. The little individual bottles are best, so that each one can remain contained and free of contaminants, rather than opening a gallon of juice at a time.


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InvisibleDickhead
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Re: Apple juice liquid culture (w/ Pics) [Re: Mushouse]
    #11047458 - 09/12/09 07:04 PM (7 years, 6 months ago)

Liquid cultures don't seem very fussy IME. Honey, Potato broth, Karo..etc.

Not surprised that Apple juice works.


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Invisible13shroomsM
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Re: Apple juice liquid culture (w/ Pics) [Re: Dickhead]
    #11047560 - 09/12/09 07:29 PM (7 years, 6 months ago)

is this your first apple juice LC or do you use them frequently?

I havent got an LC to do any good with the exception of honey.:shrug:


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Invisibletruskool
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Registered: 06/28/09
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Re: Apple juice liquid culture (w/ Pics) [Re: 13shrooms]
    #11047565 - 09/12/09 07:30 PM (7 years, 6 months ago)

Quote:

13shrooms said:
is this your first apple juice LC or do you use them frequently?

I havent got an LC to do any good with the exception of honey.:shrug:



Same here and the only way it works is if it is a grain LC


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Invisible13shroomsM
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Re: Apple juice liquid culture (w/ Pics) [Re: truskool]
    #11047618 - 09/12/09 07:41 PM (7 years, 6 months ago)

yeah, grain LC is more of a myc transfer, by far my fav LC tek:headbang3:


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Invisibletruskool
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Re: Apple juice liquid culture (w/ Pics) [Re: 13shrooms]
    #11047634 - 09/12/09 07:43 PM (7 years, 6 months ago)

Quote:

13shrooms said:
yeah, grain LC is more of a myc transfer, by far my fav LC tek:headbang3:



yea works beautifully for me, but I am going to try this for sure.  I like simplicity


--------------------
Ask  AMU    for    the  best    mycology    advice  out  there

  Roll it :burnone:While I troll it.  :maximumtrolling:

I don't mean to boast, but dam if I don't brag.


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Offlinenexus1946
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Re: Apple juice liquid culture (w/ Pics) [Re: Mushouse]
    #11048007 - 09/12/09 09:09 PM (7 years, 6 months ago)

Quote:

Mushouse said:

It was a 6-pack of Minute Maid little 10oz apple juice bottles. The package said "100% Juice."

Ingredients: sure filtered water, concentrated apple juice, less than 0.5% of: vitamin C (ascorbic acid), calcium citrate, potassium phosphate

The generic grocery store brand 6-pack would have been used, because they are identical juice, but the Minute Maid bottles were bought for their shape, as the original plan was using them right in their own bottles.

The apple juice was not pressure cooked or treated in any way. Used straight out of the bottle. The little individual bottles are best, so that each one can remain contained and free of contaminants, rather than opening a gallon of juice at a time.




Thanks for the info. :grin:

I think I may try this with a grain LC.


--------------------
Gypsum/Drywall Tek
The Story Of Russell The Texas (Cube) Bear



The human race's prospects of survival were considerably better when we were defenceless against tigers than they are today when we have become defenceless against ourselves.
-Arnold J. Toynbee


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