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#21 PolarDBN

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Posted 31 August 2018 - 05:40 PM



Mmmm my strategy of multi-quote is not going to work here.



Interesting, how fast are we talking? As the isolates I've taken thus far seem to be a bit on the slow side. Maybe that's just because I'm used to cubes? The cultures cabinet is a little on the cool side at the moment, around 22C (I'm sure they'd appreciate another 2C at least). I do believe the grain I have at the moment is in fact WBS.


This is Pan cam Goliath 3 days in, tall baby food jar, probably about 50% there. Cyans if I remember was like a week also.

WBS + tiny bit of brown mustard seeds + some gypsum, 24 hour soaked. Probably don't need all the additions. Just can't follow recipes. ;)

The small seed make for great spawn. Pans don't seem to enjoy shaking. 091418e62b4b6f8cba8a2d6e600fc8ce.jpg

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#22 425nm

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Posted 31 August 2018 - 05:54 PM

Hmmm, I know slow growers like Shiitake prefer denser substrates (why they do so well on enriched sawdust) and at least part of the reason for that is because they mycelia doesn't have to grow as far to reach the next morsel of food. Perhaps I should go on a quest for finer grains. Millet or bird seed perhaps? Depending on what is cheapest.
Your WBS looks different than mine. I'll have to check exactly what it is that I have. I could have sworn it was whole wheat kernels.

I am definitely not seeing week colonization times.
Those 6 1L jars mention in the first post still aren't showing any growth (no Pans no mold no bacteria no nothing). Its very odd, I actually wonder if the fermentation of the grain altered the pH or created some inhibitory by product.



#23 Sidestreet

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Posted 01 September 2018 - 06:41 PM

I've got my first glass petri no-pours on the stove right now and all of this is super helpful. 

 

 

When I've done this in the past I had a lot of trouble with them boiling over and getting agar all over them. Any tips on how to do it without the agar rebelling and escaping the Petri's? Are yours the usual 15mm high or extra deep?

 

 

Found that the internal pressure in the pressure cooker needs to decrease slowly and naturally to atmospheric. I cover mine with insulation.

When it hits atmospheric pressure (need a pressure gauge to verify) it's best to open the valve so the pc doesn't create a vacuum... And let it cool naturally from there.

 

Good call with the insulation to slow down the cooling, especially because I don't have much water in the PC with these petris.  It will try to cool down fast.

 

 

I am glad you are cooling that down slowly. It has the potential for quite a mess. You can actually lower your burner to warm to keep it from cooling down too fast also and cover the top with a towel. It usually takes about an hour to zero out the pressure (depending on your stove) and from there you can turn off your burner. It is a much slower cool down and avoids a vacuum scenario. Doing this also keeps your bags from blowing out sidewalls.

 

Lowering the burner to warm to keep it from cooling down too fast--that's a great tip.  You should put it in your myco hacks thread!

 

 

That is only part of the equation though: Don't fill your dishes past half way

 

Another good one.  On my first dishes I thought the mycelium would eat through the whole thing but it really just stays on the surface.  For this batch I poured just enough into each dish to cover the bottom.


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#24 PolarDBN

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Posted 01 September 2018 - 11:05 PM

Mmmm my strategy of multi-quote is not going to work here.



Interesting, how fast are we talking? As the isolates I've taken thus far seem to be a bit on the slow side. Maybe that's just because I'm used to cubes? The cultures cabinet is a little on the cool side at the moment, around 22C (I'm sure they'd appreciate another 2C at least). I do believe the grain I have at the moment is in fact WBS.

This is Pan cam Goliath 3 days in, tall baby food jar, probably about 50% there. Cyans if I remember was like a week also.

WBS + tiny bit of brown mustard seeds + some gypsum, 24 hour soaked. Probably don't need all the additions. Just can't follow recipes. ;)

The small seed make for great spawn. Pans don't seem to enjoy shaking. 091418e62b4b6f8cba8a2d6e600fc8ce.jpg

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Day 5 wbs. In my option, as inoculant these are ready to go. ab071c98338811e1e63f0a569f373187.jpg

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#25 425nm

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Posted 02 September 2018 - 02:34 AM

Righty-o!
I got me a big ol'bin of millet!

IMG 20180901 233702

I have also inoculated one of an LC
 

IMG 20180901 220921
 
As well as doing another transfer off of the spore germination plate. A4.
IMG 20180901 220854

 

 

@PolarDBN - Sorry if you mentioned this before but how are you inoculating those jars? Five days is very speedy.


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#26 PolarDBN

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Posted 02 September 2018 - 07:02 AM

That's a whole lot of millet, glad we got you inspired ;)

One just needs a small agar wedge to inpregnate the seeds. Shaken only at inoculation and then left to colonize. You'll have spawn in no time.

Would love to see some updates as the agar culture moves along... Should also be really quick.

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#27 onediadem

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Posted 02 September 2018 - 01:38 PM

I always use at least 5 pieces of agar. I want it to run as fast as possible. Then I go grain to grain with that. The more inoculation points the better.


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#28 425nm

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Posted 02 September 2018 - 04:31 PM

Continuing with my not exactly updates.
Those 6L of gain I shot with LC (the ones that fermented a bit before hand) are still showing ZERO signs of growth. I'm wondering if I should just pitch them and reclaim the jars or try reinoculating them?
 

IMG 20180902 133742

 

I might get some millet prepped tomorrow. Sadly I don't think I'll have any mushroom time today.



#29 jkdeth

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Posted 02 September 2018 - 05:14 PM

I'd probably toss. Mostly because every time I try to fix things I end of in a big cycle of grossness and wasted time.

That being said if you have a big ole jug of lc you aren't really risking much...

This is a known good lc right?

Trying to get my head around how fermenting would alter the grain in such a fashion as to prevent growth. Obviously yeast would have been killed in the pc, as would I believe any lactibacillus if that were even present. Hard to believe there would be even alcohol conversion to cause a problem.

#30 425nm

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Posted 02 September 2018 - 05:41 PM

I'd probably toss. Mostly because every time I try to fix things I end of in a big cycle of grossness and wasted time.

That being said if you have a big ole jug of lc you aren't really risking much...

This is a known good lc right?

Trying to get my head around how fermenting would alter the grain in such a fashion as to prevent growth. Obviously yeast would have been killed in the pc, as would I believe any lactibacillus if that were even present. Hard to believe there would be even alcohol conversion to cause a problem.

I do have plenty of LC plus a new LC colonizing as I type. The LC in question I tested the week before on a small 120mL jar of grain and it worked no problem.
I know right? Its odd. I can think of a few ways:

 

A. Change the pH - the yeast and lactobacillus would generate a bunch of byproducts as they breakdown the available starches and sugars HOWEVER I did put gypsum in the soak water plus I rinsed the crap out of the grain (I can't imagine they would have been growing much deeper than the surface of the grain).

 

B. Change the nutritional content - it is possible that they ate up a large amount of the available sugars & starches? I have a hard time believe they could have broken them all down in a 30-36 hour period.

C. Some byproduct of the fermentation is preventing the Pans from growing however the rinsing should have removed most of it.

I suppose its possible that the grain hadn't cooled off as much as I had thought at the core of the jars? I did PC them and inoculate on the same day but I could handle the jars without any sensation of warmth plus I always shoot down the side of the jars so I can see if colonization takes hold.



#31 jkdeth

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Posted 02 September 2018 - 09:19 PM

Yeah, rinsed, then pc'd you'd think would take care of any issues. I could see the process creating a heavier bacteria load, just from being moist for a period of time, but the pc should have taken care of that. You'd think think a fungus that loves poo wouldn't be picky about gently used grain.

#32 PolarDBN

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Posted 02 September 2018 - 10:29 PM

I would rather blame the LC. If you get the WBS going soon you could try use that to expand to these. Bet you a 50 the colonize.



#33 Cigarsam

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Posted 03 September 2018 - 04:13 AM

If your conditions are right, which form what I am catching up on your in good hands here.
Peacefrog and his Pan prints I think are very unique. (Upper Echelon)
Some of your spores might even fruit on your agar plates.
All the best with Pan Hawaiian adventure.

Edited by Cigarsam, 03 September 2018 - 04:24 AM.

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#34 425nm

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Posted 04 September 2018 - 10:27 PM

The A4 has fuzzed up nicely

 

IMG 20180904 185131
 
IMG 20180904 185136

 

The G2G I did with the proof jar of the old Pan LC are colonizing nicely in some wheat.

 

IMG 20180904 191609
 
IMG 20180904 191621

 

Which makes the case of the six still uncolonized 6L of wheat all the more puzzling. I even shook one a few days ago to check for contamination/encourage a bloom of any that might be hiding and not even that has shown itself. On that note I think I'm going to re-sterilize and reinoculate that grain. But that is a task for tomorrow ) hopefully.

 

Today I made some more agar, sterilized needles, and slants!

 

IMG 20180903 213434
 
The agar is just MEA and slants are the conrmeal glucose (aka. dextrose) media describe by Stamets (also in FastFred's cookbook). Low nutrient, should be good for storage. I think a project to make a cool little slant case is in order!

The needles are for some test jars of millet I have in the pressure cooker as I type this.

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#35 RamenShaman

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Posted 05 September 2018 - 09:14 AM

It seems I am a little late to this thread I have to agree That the the Cope Cyan Hawaiians that I have been working with are very quick to colonize but I have never had any ill affects from shaking like DBN has, my cultures generally start recovering by the next day.

I have also used WBS that has soaked a couple extra days more than I planned but after rinsing and pressure cooking had no trouble with it I am surprised to see your large grain jars inoculated with LC are not growing. I find this quite odd...

I’ll have to pay more attention to this thread

Good luck and good vibes bud

Edited by RamenShaman, 05 September 2018 - 09:14 AM.


#36 425nm

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Posted 06 September 2018 - 08:34 PM

So I shot some test jars of millet. First how I prepped the millet, I did exactly what I do with wheat. Pour boiling water over it in the jars with some gypsum, closed, and periodically shake to prevent the swelling immobilizing the grain. A few guides on the internet led me to believe this would work. A lot of folks seem to do a fill with specific volume to millet and PC in one shot which I'm sure works but I'd rather have an opportunity to wash away excess starch.
Anyway, back to the grain at hand. They kinda swelled up? But not as much as I expected. I'm not sure they've actually taken on enough water? I also shot the jars with 4mL of inoculum from the older LC. I realize this is a bit much LC for the size of jar.

 

IMG 20180906 160259
 

It seems unlike wheat the millet doesn't readily soak up excess liquid (see small jar in above picture) all that well which is a bit problematic. Now that liquid can be relatively easily redistributed via shaking after inoculation which I prefer not to do (if the jars come up contaminated it makes it hard to know if the grain didn't sterilize properly vs the LC being contaminated).

 

Does anyone have a millet prep they'd recommend vs my soak in the jar with boiling water?



#37 onediadem

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Posted 06 September 2018 - 09:40 PM

If you do try those same jars again make sure to rehydrate them because they look super dry to me. And, there doesn't seem to be any condensation on the jar whatsoever.



#38 425nm

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Posted 06 September 2018 - 11:50 PM

If you do try those same jars again make sure to rehydrate them because they look super dry to me. And, there doesn't seem to be any condensation on the jar whatsoever.

 

The wheat jars? or the millet?



#39 onediadem

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Posted 07 September 2018 - 12:48 AM

The ones you said you wanted to try to use again were these right? They are the ones that look too dry to me.

 

sml_gallery_149784_1648_835806.jpg



#40 425nm

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Posted 07 September 2018 - 08:58 PM

IMG 20180907 181141

Cracked the jars open. No obvious no of either pan mycelia nor contamination growth. Still smelled slightly of fermentation but mostly of moist grain.

Re-hydration and re-sterilization is underway!

 

IMG 20180907 184435
 
IMG 20180907 184505

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