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hamloaf
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Unconventional Inoculation of Large Spawn Bags with Spore Solution. 1
#21514321 - 04/07/15 10:13 AM (9 years, 1 month ago) |
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Inoculating sterile grain substrates is a less than ideal way to germinate spores. Inoculating large grain substrates in bags with spore solution is an even lesser than ideal way to germinate spores in. The reason(s) being is that spores have a germination period of up to 14 days. Sometimes longer. Sometimes shorter. Give, or take five days or so in each direction.
The reason we sterilize our grain substrates is to give our mushroom mycelium a head start against the onslaught of competitor mold and bacteria spores that are in direct competition for the same nutrient sources, and environmental conditions that the mushroom mycelium favors on indoor mushroom cultivation. There is actually a very fine line, and dance between favorable conditions for molds and mushroom mycelium that must be waltzed in order to succeed.
Keep in mind that grain substrates have to remain sterile longer when you inoculate them with spores, because not only do they have to support growth while the spores are germinating. Once germination occurs the substrates must then be able to maintain sterile equilibrium through out the mycelium running phase.

Was gifted a full 10cc's of Brazilian cubensis spores in a syringe from a generous Shroomery member, and didn't feel like doing the whole agar thing despite having the plates poured + there was an extra couple of bags left over from production run comprised of 2 gallons of sterilized whole oats, so figured why not, and inoculated them each with 5 full cc's of spore syringe solution. Just flame sterilized the needle, stuck the needle through the bags and sprayed spore solution all over the sterile grain substrates, laughing the whole way. 

Since the substrates are so large, and crammed together in the PC for sterilization, they were sterilized at 20-22 psi's for 4 hours. Also, if the syringe ends up being contaminated, than the only things lost are about 1 dollars worth of substrate materials, and the hours that it took to prepare the sterile grain substrates.
Update 4-21-2015
2 weeks from inoculation, and no contams, no myce, no nothing.

So the rest of this LC (LC featured in the Bulk LC thread)

Was placed onto the stirplate, and stirred for 10-20 minutes @ 555 rpm's while surfaces of the lab were wiped down with an alcohol soaked paper towel.

Next, the culture was extracted from the depths of the quart jar with the trusty 4" long, 12 gauge blunt-dispensing needle making 2 - 10 cc syringes full of nice, thick LC.


Each bag that was inoculated 2 weeks ago with spore syringe solution that has done nothing was then blasted with a full syringe's contents, and sealed with clear mover's tape placed right over the injection hole.

Now let's see what's really going on. 
Update 4-25-2015.
First signs of recovery were noticed from the LC onto the grains just four days out from inoculation.

Update 5-5-2015.
Here is the mycelium recovering from being shook about 5 days ago.

Inoculm used on those bags is the same LC master that was used to inoculate the bags in the Bulk LC Thread.
Update 5-14-2015.
Bags featured in original post that were originally inoculated with dead spores, then 2 weeks later inoculated with a suspect LC are now fully colonized.

The plans for these bags are to throw them into the regularly scheduled production run cycle. This means each 10 quart bag of fully colonized cubensis grain spawn will be launched into a bulk substrate comprised horse manure/coir/vermiculite/gypsum inside of 50 quart, clear plastic storage totes modified into monotub fruiting chambers, and fruited inside of the ol' humidity controlled environment. Each bag will yeild a total of 5 monotubs of the aforementioned size with a substrate depth of 1-2 inches, spawned at a 1:2 grain spawn to bulk substrate ratio.

Edited by hamloaf (05/14/15 11:07 PM)
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bw86
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Re: Unconventional Inoculation of Large Spawn Bags with Spore Solution. [Re: hamloaf]
#21514418 - 04/07/15 10:36 AM (9 years, 1 month ago) |
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You getting bored over there?
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hamloaf
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Re: Unconventional Inoculation of Large Spawn Bags with Spore Solution. [Re: bw86]
#21514490 - 04/07/15 10:50 AM (9 years, 1 month ago) |
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Just utilizing what was laying around to generate something out of nothing, and thought I would share.
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13shrooms
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Re: Unconventional Inoculation of Large Spawn Bags with Spore Solution. [Re: bw86] 2
#21514561 - 04/07/15 11:05 AM (9 years, 1 month ago) |
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back in the day whyblameus used to inoc 4qt bags of grains with whole syringes of spore solution, as long as you dont throw off the moisture content of the grains things work out just fine. 
https://files.shroomery.org/files/10-15/156936370-IMG_2765.jpg
1 syringe inocd 1 bag spawned to 1 mono 
I start with 1 grain jar then g2g to 1 bag then spawn to a mono..
the only issue ive ran into this method is if you dont use enough spore solution the grains will absorb the moisture making conditions crap for germination..
so using alot of solution seems to help in this case but not ideal for normal germination, less is more normally 
another informative post by Ham
--------------------
A M U Click here ^ for the AMU forum VVV AMUs Free Active/Edible/Exotic Spore Print or Syringe or Edible Culture Trade Thread VVV "Man is the sex organ of the machine world" ~ Marshall McLuhan
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blackdust


Registered: 02/28/09
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Re: Unconventional Inoculation of Large Spawn Bags with Spore Solution. [Re: 13shrooms]
#21537359 - 04/12/15 01:21 PM (9 years, 1 month ago) |
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Ham is so bad ass Who else posts such good content on a weekly basis? Keep up the good work bro
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hamloaf
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Re: Unconventional Inoculation of Large Spawn Bags with Spore Solution. [Re: blackdust]
#21578430 - 04/21/15 10:01 PM (9 years, 28 days ago) |
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2 weeks from inoculation, and no contams, no myce, no nothing.

So the rest of this LC (LC featured in the Bulk LC thread)

Was placed onto the stirplate, and stirred for 10-20 minutes @ 555 rpm's while surfaces of the lab were wiped down with an alcohol soaked paper towel.

Next, the culture was extracted from the depths of the quart jar with the trusty 4" long, 12 gauge blunt-dispensing needle making 2 - 10 cc syringes full of nice, thick LC.


Each bag that was inoculated 2 weeks ago with spore syringe solution that has done nothing was then blasted with a full syringe's contents, and sealed with clear mover's tape placed right over the injection hole.

Now let's see what's really going on.
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blackdust


Registered: 02/28/09
Posts: 8,327
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Re: Unconventional Inoculation of Large Spawn Bags with Spore Solution. [Re: hamloaf]
#21579610 - 04/22/15 09:17 AM (9 years, 28 days ago) |
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Thank you!!!!
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KindofBlue
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Re: Unconventional Inoculation of Large Spawn Bags with Spore Solution. [Re: blackdust]
#21579716 - 04/22/15 09:49 AM (9 years, 28 days ago) |
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Nice
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hamloaf
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Re: Unconventional Inoculation of Large Spawn Bags with Spore Solution. [Re: KindofBlue]
#21596941 - 04/25/15 09:22 PM (9 years, 24 days ago) |
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First signs of recovery were noticed from the LC onto the grains just four days out from inoculation.
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Buck513

Registered: 04/17/14
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Re: Unconventional Inoculation of Large Spawn Bags with Spore Solution. [Re: hamloaf]
#21597626 - 04/26/15 01:25 AM (9 years, 24 days ago) |
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-------------------- Fail to plan and you plan to fail. Enter the Ban Lottery
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eatyualive
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Re: Unconventional Inoculation of Large Spawn Bags with Spore Solution. [Re: Buck513] 1
#21599005 - 04/26/15 12:40 PM (9 years, 24 days ago) |
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hamloaf
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Re: Unconventional Inoculation of Large Spawn Bags with Spore Solution. [Re: eatyualive]
#21640310 - 05/05/15 05:03 PM (9 years, 15 days ago) |
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Here is the mycelium recovering from being shook about 5 days ago.

Inoculm used on those bags is the same LC master that was used to inoculate the bags in the Bulk LC Thread.
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hamloaf
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Re: Unconventional Inoculation of Large Spawn Bags with Spore Solution. [Re: hamloaf]
#21680532 - 05/14/15 08:11 PM (9 years, 6 days ago) |
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Bags featured in original post that were originally inoculated with dead spores, then 2 weeks later inoculated with a suspect LC are now fully colonized.

The plans for these bags are to throw them into the regularly scheduled production run cycle. This means each 10 quart bag of fully colonized cubensis grain spawn will be launched into a bulk substrate comprised horse manure/coir/vermiculite/gypsum inside of 50 quart, clear plastic storage totes modified into monotub fruiting chambers, and fruited inside of the ol' humidity controlled environment. Each bag will yeild a total of 5 monotubs of the aforementioned size with a substrate depth of 1-2 inches, spawned at a 1:2 grain spawn to bulk substrate ratio.
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takofako
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Registered: 01/05/15
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Re: Unconventional Inoculation of Large Spawn Bags with Spore Solution. [Re: hamloaf]
#21680630 - 05/14/15 08:32 PM (9 years, 6 days ago) |
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Love your planet
-------------------- Sorry about spelling, english is not my main language My Grow Log and My Teks
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