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eatyualive
Eat's You Alive :)



Registered: 08/17/01
Posts: 19,026
Loc: In Your Head
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Re: Slurry Spawn or Liquid Inoculant [Re: blackout]
#21008397 - 12/21/14 05:06 PM (9 years, 6 months ago) |
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ha blackout. always chiming in with some killer ideas.
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blackdust


Registered: 02/28/09
Posts: 8,327
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Re: Slurry Spawn or Liquid Inoculant [Re: blackout]
#21008447 - 12/21/14 05:17 PM (9 years, 6 months ago) |
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Quote:
Step 4. The Eberbach stirrer is placed on the power unit and stirred in 3-second bursts. (The blender I use rotates at 8400 rpm.) Pausing for 5 seconds, the surviving chunks of agar fall downwards into the blades. Another 3-second burst decimates these pieces. One more 5-second pause is followed by the last 3-second, high-speed stir. In effect, the stirring process has created thousands of chopped strands of mycelium, in short cell chains.
Step 5. The water/mycelium blend is transferred, 250 ml. at a time in equal proportions, into the three 1500 ml. Erlenmeyers. A remote syringe, pipette, or liquid pump can be used. Less elaborate is to simply "free-pour" equal volumes of myceliated fluid from the Eberbach into each Erlenmeyer. The non-absorbent cotton stoppers are, of course, removed and replaced with each pouring, being careful not to allow contact between the cotton stopper and any contaminated surface. Each Erlenmeyer is placed on stir plates or on a shaker table and rotated at 100-200 rpm. for 48-72 hours. The water broth is continuously stirred to allow transpiration of metabolic gases and oxygen absorption. The fluid has a milky-brown color and is not translucent. Settling of the heavier components is clearly visible when the stirring process is interrupted. Upon completion, 3000 milliliters of mycelium are rendered in liquid form. The hyphae, recovering from the damage of being cut by the spinning blades of the blender, are stimulated into vigorous re-growthAt a point several cells away from the cut ends, nodes form on the cell walls, new buds push out, and branch. A vast, interconnected fabric of cells, a mycelial network, forms. The branches fork continuously. After two to four days of re-growth in the nutrient enriched broth, each Erlenmeyer flask GENERATING GRAIN SPAWN 149 becomes its own universe, hosting thousands of star-shaped, three dimensional colonies of mycelium. This is the stage idealfor inoculation into sterilized substrates, especially in the generation of grain spawn masters. (See Figure 120). Far more bioactive than the same mycehum transferred from the two-dimensional surface of a petri dish, each hyphal cluster grows at an accelerated rate subsequent to transfer to the grain media. If, however, the liquid media is not used at its peak rate of growth, and stirs for nearly a week, the colonies lose their independence and coalesce into a clearly visible contiguous mycelial mat. Long mycelial colonies adhere to the interface of the fluid surface and the inside of the flask. Chains of mycelium collect downstream from the direction of rotation. Soon after their appearance, often overnight, the media becomes translucent and takes on a rich amber color. A large glob of mycelium collects on the surface and can be mechanically retrieved with a pair of tweezers, forceps, or scalpel, if desired. The remaining clear amber fluid contains super-fine satellite colonies and hyphal fragments. Bypassing the fluid through a microporous filter, the mycelium can be recaptured. This technique is especially attractive for those whose goal is running tests on small batches of myceium. With many species I have grown, the conversion ratio of sugar/wood to mycehium (dry weight) approaches 20%. This percentage of conversion is nearly 80% biological efficiency, considered good in the commercial cultivation of gourmet mushrooms.
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Dilated
KB Cubensis Fanaticus



Registered: 03/28/14
Posts: 2,347
Loc: The Ether
Last seen: 3 years, 7 months
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Re: Slurry Spawn or Liquid Inoculant [Re: blackdust]
#21011347 - 12/22/14 08:48 AM (9 years, 6 months ago) |
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stareatclouds
star eat clouds?



Registered: 09/29/14
Posts: 10,329
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Quote:
MustardTiger66 said: I made some LI from a fully colonized agar plate and it's day two and I see nothing in my oat jars. I wonder if the blade destroyed the myc?
I shake the shit out of my LI to distribute it throughout and I usually see growth maybe 3-4 days after. I know it takes a bit for the myc to start colonizing again because the blades do fuck it up for a bit. How is your growth now, MT?
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TheMustardTiger


Registered: 08/19/13
Posts: 4,700
Loc: Sunnyvale Trailer Park
Last seen: 6 years, 10 months
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Re: Slurry Spawn or Liquid Inoculant [Re: stareatclouds]
#21019592 - 12/23/14 11:11 PM (9 years, 6 months ago) |
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Quote:
stareatclouds said:
Quote:
MustardTiger66 said: I made some LI from a fully colonized agar plate and it's day two and I see nothing in my oat jars. I wonder if the blade destroyed the myc?
I shake the shit out of my LI to distribute it throughout and I usually see growth maybe 3-4 days after. I know it takes a bit for the myc to start colonizing again because the blades do fuck it up for a bit. How is your growth now, MT?
It recovered. I just did another batch of WBS so we'll see how that goes.
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stareatclouds
star eat clouds?



Registered: 09/29/14
Posts: 10,329
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Sweet. Mine is coming along great after about 4 days now as well. LI and slurries are awesome. Going to buy some more BRF soon so I can do some more!
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eatyualive
Eat's You Alive :)



Registered: 08/17/01
Posts: 19,026
Loc: In Your Head
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Re: Slurry Spawn or Liquid Inoculant [Re: stareatclouds]
#21021008 - 12/24/14 11:29 AM (9 years, 6 months ago) |
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ok so here is an updated to the grain slurry tubs. this is my first few attempts at it and i have no contamination on 3 tubs going into 1st flush. the two tubs that i did 1/2 quart grain slurries to. the moisture content is too low in all the tubs i mixed up. so 2 didn't colonize the substrate completely. both are pinning now. i misted the uncolonized coir and it acted like a casing layer. so, check it out. no contams so far.

the 1 quart slurry is now pinning.
  
so not too shabby. im pretty sure i reduced the moisture too much. im going to make some adjustments next trial run.
updating this post..
http://www.shroomery.org/forums/showflat.php/Number/21008247#21008247
Edited by eatyualive (12/24/14 11:31 AM)
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blackdust


Registered: 02/28/09
Posts: 8,327
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Re: Slurry Spawn or Liquid Inoculant [Re: eatyualive]
#21021042 - 12/24/14 11:35 AM (9 years, 6 months ago) |
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Get em Eat!
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TheMustardTiger


Registered: 08/19/13
Posts: 4,700
Loc: Sunnyvale Trailer Park
Last seen: 6 years, 10 months
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Re: Slurry Spawn or Liquid Inoculant [Re: blackdust]
#21021049 - 12/24/14 11:36 AM (9 years, 6 months ago) |
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I'll take a healthy, partially colonized sub over a stalled contam sub any day of the week. Looking forward to the results
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eatyualive
Eat's You Alive :)



Registered: 08/17/01
Posts: 19,026
Loc: In Your Head
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yeah i need to adjust the moisture content to field capacity on the next trial. im going to try and get an exact recipe to make it easier.
also i think the reason the wbs works better is that the particle size is smaller so your only getting a percentage of the grains being pulverized where if your using larger grain like rye berry or oats the size of the particle that is uncolonized and exposed. the wbs seems to minimize this.
Edited by eatyualive (12/24/14 11:42 AM)
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MudaFuka
Poppin bottles



Registered: 12/14/13
Posts: 18,648
Loc: Canada
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Re: Slurry Spawn or Liquid Inoculant [Re: eatyualive]
#21021165 - 12/24/14 12:00 PM (9 years, 6 months ago) |
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blackdust


Registered: 02/28/09
Posts: 8,327
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Re: Slurry Spawn or Liquid Inoculant [Re: azur]
#21022740 - 12/24/14 08:40 PM (9 years, 6 months ago) |
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I think that slurry may be contam resistant considering these tins were made with slurry in open air and the tins were only oven pasteurized.
      
Warning: These were not created with amazing results in sight. I was looking to see how contam resistant this method is for me.
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eatyualive
Eat's You Alive :)



Registered: 08/17/01
Posts: 19,026
Loc: In Your Head
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Re: Slurry Spawn or Liquid Inoculant [Re: blackdust]
#21040018 - 12/29/14 03:31 PM (9 years, 5 months ago) |
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ok so i had to go out of town during the harvest and came back to some 2 day overgrown fruits with plenty of spores. but hell it worked!
all 3 tubs fruited. the tray on the right is the 1 quart wbs grain slurry and the tray on the left is the 2 1 pint wbs grain slurries. i dunked the 1 quart grain slurry tub for a 2nd flush and i chunked the 2 1 pint slurry tubs to make room for some new tubs.

so far no contams. lets see what happens on 2nd flush. updated log. ill update weights soon. im sure its nothing worth while. but i think the 2 quart slurry should do well!
http://www.shroomery.org/forums/showflat.php/Number/21008247#21008247
Edited by eatyualive (12/29/14 03:41 PM)
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MudaFuka
Poppin bottles



Registered: 12/14/13
Posts: 18,648
Loc: Canada
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Re: Slurry Spawn or Liquid Inoculant [Re: eatyualive]
#21040026 - 12/29/14 03:33 PM (9 years, 5 months ago) |
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That looks like hell but I'm glad to see it worked out for you.
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,887
Loc: Canada
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Re: Slurry Spawn or Liquid Inoculant [Re: MudaFuka]
#21040029 - 12/29/14 03:35 PM (9 years, 5 months ago) |
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Very interesting, nice work
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TheMustardTiger


Registered: 08/19/13
Posts: 4,700
Loc: Sunnyvale Trailer Park
Last seen: 6 years, 10 months
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Re: Slurry Spawn or Liquid Inoculant [Re: MudaFuka]
#21040032 - 12/29/14 03:35 PM (9 years, 5 months ago) |
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Spore bombbbbbbbbbbbb.
Nice job as always, eat
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eatyualive
Eat's You Alive :)



Registered: 08/17/01
Posts: 19,026
Loc: In Your Head
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Re: Slurry Spawn or Liquid Inoculant [Re: MudaFuka]
#21040037 - 12/29/14 03:37 PM (9 years, 5 months ago) |
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yeah it wanted to fruit the day i was going out of town. oh well. lol i was going to chunk all the fruits anyway. well check this one out muda.
this is a 2 quart grain slurry tub that colonized in 3 days!
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eatyualive
Eat's You Alive :)



Registered: 08/17/01
Posts: 19,026
Loc: In Your Head
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Quote:
MustardTiger66 said: Spore bombbbbbbbbbbbb.
Nice job as always, eat 
thanks. yeah and now i got fam in town. trying to work in some experiments between family in town lmao.. busy busy.
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TheMustardTiger


Registered: 08/19/13
Posts: 4,700
Loc: Sunnyvale Trailer Park
Last seen: 6 years, 10 months
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Re: Slurry Spawn or Liquid Inoculant [Re: eatyualive]
#21040042 - 12/29/14 03:39 PM (9 years, 5 months ago) |
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Quote:
eatyualive said: yeah it wanted to fruit the day i was going out of town. oh well. lol i was going to chunk all the fruits anyway. well check this one out muda.
this is a 2 quart grain slurry tub that colonized in 3 days!

3 days?
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,887
Loc: Canada
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Re: Slurry Spawn or Liquid Inoculant [Re: eatyualive]
#21040049 - 12/29/14 03:41 PM (9 years, 5 months ago) |
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Quote:
eatyualive said: yeah it wanted to fruit the day i was going out of town. oh well. lol i was going to chunk all the fruits anyway. well check this one out muda.
this is a 2 quart grain slurry tub that colonized in 3 days!

I got me 5 quarts of colonized spawn I was going to spawn today
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