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Pink's Bulk Adventures


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#1 PinkMenace

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Posted 02 December 2017 - 10:42 PM

Here is a road map for my first bulk adventure.

 

Planning

 

Steeping:  Cue's WBS Steep Tek

Jars: 1 quart jars with lids I made earlier in this thread - filling 3/4ths full of the WBS. I'm aiming for 7 jars, hoping that 4 will survive.

Sterilization: 90 min @ 15 psi

Inoculation: Syringes I made from a Redboy X print from @coorsmikey (Thank you!).

Shaking (from gruncle @microbe): "Grain i shake around 25% then at 75% then a final shake 24 hours before i milk or expand. The last shake isnt to check for contamination because if its in there after the 2nd shake, evidence of any contamination will present itself. It can be slow recovery, odor, or isolated areas where the myc is avoiding and wont always see the actual contamination. My final shake is so that it begins to recover before I expand because I shake the jars down like they owe me money and its vicious.....much for vicious then any g2g or mycelium milking." + I'll shake when I first inoculate. 

Substrate and fruitingCoco Coir tek by Wharfrat (minus coffee) 

 

And so the story begins...

 

I started by bringing down my 20lb bag of Wild Bird Seed. You will notice that it contains cracked corn. Cracked corn is high in starch which can create a film that inhibits colonization and is quite delicious to other microbes (according to @microbe). This can however be overcome by thorough rinsing before and after hydration (the steep).

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I got a mixing bowl and shoveled WBS into it, aiming to fill 5-7 quart jars 3/4ths full. I filled a jar 3/4ths full and dumped it in 5 times, assuming there would be expansion (though I may have been scooping over the 3/4th point).

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My next plan was to fill the bowl with water so that the sunflower seeds floated allowing me to scoop them out. However my mixing bowl was too small (better start bigger next time) so I upgraded to small brew kettle.

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And SURPRISE! The sunflower seeds were not the only thing that floats down here. I had to get a strainer and laboriously rub and water until the non-sunflower seeds were washed out. Pretty soon I ran out of head room, and had to focus on rubbing.

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After I removed as many sunflower seeds as I could stomach, I put a grain bag over the kettle and very carefully dumped the contents.

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Next I rinsed it thoroughly, including laying it flat and running water over it while massaging it.

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Next I experimentally submerged the water in the kettle, seeing how much it would take to submerge it. I then dumped out that water and replaced it with fresh water, putting in a lot extra since I did not know how much the WBS would absorb (according to Cat's it wouldn't absorb much, but I didn't want to risk it). The photo shows before I added the extra.

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I brought the water to boil and went and hung out in Mycotopia chat. If you're interested in chatting with us or just want to help Mycotopia keep running, sign up for VIP by looking into the "Go VIP!" menu at the top of every page! 

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After the water started boiling, I turned off the heat and put the grain bag in. I then put the lid back on.

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I let it steep for 90 minutes and then took the grain bag out and put it on a grate. When I came back to check on it in an hour and lifted it, a torrent of water came pouring out. I ended up shaking the bag up and down, leaving it alone for a bit, and then repeating until no water came out. In the future I will do that from the start.

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I dumped the now hydrated bird seed into a bowl and got a ladle to start ladling out WBS into jars.

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I leaned towards too little and away from too much when filling them and finally came out with 7 jars filled to often less than 3/4ths. Oh well. I figure it'll be easier to shake.

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Lastly I threw on some tinfoil hats so that the pressure cooker couldn't read their thoughts and am currently cooking them at 15PSI for 90 minutes.

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Edited by PinkMenace, 02 December 2017 - 11:32 PM.

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#2 Jeepster

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Posted 02 December 2017 - 11:20 PM

Wow, epic thread, PM, very nice. Great photos, too! Thread like this will help new folks.
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#3 wharfrat

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Posted 02 December 2017 - 11:57 PM

love the added touch with the star  :biggrin:  pulling up a chair


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#4 PinkMenace

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Posted 03 December 2017 - 12:06 AM

There's more about sunflower seeds being bad and why here. And an experiment to illustrate it in Myc's Sunflower Seed/ Trash experiment

 

There is also the following fun tidbit from dragonstud725 in the first thread, presenting how they made their own sieve to get them and corn out:

 

 

I use the cheap wally world wbs works great. What I do about the black sunflower seeds and the very few pieces of corn that are in a bag is I bought a cheap plastic strainer from the dollar tree then drilled out the holes so all the wbs we use falls threw into a bowl underneath while keeping the big black sunflower seeds in the strainer. It works great and gets rid of all the big black sunflower seeds and just leaves a few pieces of the crushed sunflower seeds in the bowl and they are easily picked out of the bowl now I can get all the sunflower seeds out of a whole bag in no time at all. 

 

I am tempted to try this with a drill and a small plastic bin - I don't see a reason to start with a strainer if the holes are small enough (i.e. don't need a guide). I'll post a tutorial on it if I do.


Edited by PinkMenace, 03 December 2017 - 12:07 AM.

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#5 CatsAndBats

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Posted 03 December 2017 - 11:20 AM

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Edited by CatsAndBats, 03 December 2017 - 11:20 AM.

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#6 PinkMenace

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Posted 03 December 2017 - 05:25 PM

Inoculation day!

Planning

 

Inoculation: I plan to inject 2CCs of Redbox X into each quart jar.

Procedure: I'm inoculating in my Still Air Box (SAB) I made by cutting square holes in a sterilite bin with a dremel. I'm essentially I'm using the same procedure as I do for PF. 

 

Wheee centered text!

 

I gathered all the supplies ahead of time, so I had everything handy.

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First I took an empty sample jar to make sure that the jars fit through the arm holes.

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I wiped down the inside of the SAB along with the lid

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I next sprayed inside the bin

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Then I put the lid on and sprayed inside each hole

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Then I watched Parks and Recreation to let it settle. Some people suggest 20 minutes, I got bored and didn't wait that long. Here's a shot of Ron watching TV.

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I then put my first jar in the SAB to get it ready for the next step.

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I then put some 70% rubbing alcohol on a paper towel and wiped my gloves thoroughly to sanitize them. I sanitize my gloves each time as the very last possible step before inoculating. This is a bit paranoid, I know, but I like going through the motions.

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I then got my needle red hot and injected it through the silicon injection port I had created. Unfortunately since it was so hard and I was using a blunt needle, I had to make sure it was still red hot.

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I repeated this, injecting 2CCs each time into the jar.

 

I shook the living daylights out of each of the jars in every way and direction possible. Imagine that you're trying to get a single seed to travel through every part of the jar and make friends with all the other seeds. Make sure it all moves freely and doesn't clump, as much as you can manage.

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Lessons learned:

 

* Blunt needles are really frustrating and should be avoided.

* Remember to tighten the jar lids!


Edited by PinkMenace, 03 December 2017 - 05:26 PM.

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#7 CatsAndBats

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Posted 03 December 2017 - 05:30 PM

Well done, and you're not being paranoid, you're practicing thorough aseptic technique.


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#8 425nm

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Posted 03 December 2017 - 05:54 PM

 

Well done, and you're not being paranoid, you're practicing thorough aseptic technique.

 

I second this. A mycologists aseptic technique is a microbiologists neurosis. Alcohol washing your gloves unconsciously is where you want to be. Getting down a ritual and good aseptic habits will save you from those moments when you zone out in the midst of a complex and fiddly procedure.
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#9 CatsAndBats

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Posted 03 December 2017 - 06:52 PM

 

Well done, and you're not being paranoid, you're practicing thorough aseptic technique.

 

I second this. A mycologists aseptic technique is a microbiologists neurosis. Alcohol washing your gloves unconsciously is where you want to be. Getting down a ritual and good aseptic habits will save you from those moments when you zone out in the midst of a complex and fiddly procedure.

 

 

 

Muscle memory. Tvcasualty actually suggests that one practice in one's SAB/GB/LFH with no cultures, just to get the muscle memory. It's sound advice. Once you do 1000 inoculations/agar transfers you can literally watch television [pun intended] while you work.


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#10 PinkMenace

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Posted 04 December 2017 - 04:11 PM

Note to self: the WBS swells during PC so only fill half full. I got lucky and did near half by happenstance.
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#11 PinkMenace

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Posted 08 December 2017 - 11:23 AM

Well, I got bored last night and decided to try spawning grated BRF cakes to coir in oven bags, like MLBjammer's coir-only sandbags, but inoculated with cakes. This follows the premise of the sandbag tek by sandman, where you do your fruiting in a bag. I used wharfrat's Coir/Coffee substrate tek for a guide on how to deal with coir.

 

With these four teks in hand and the twist of spawning with grated BRF cakes, I was ready to go.... And wow did I keep messing up.

 

I got my coir out and started to boil some water, measuring out half a gallon as I planned to only do have a brick of coir, cutting MLBjammer's tek in half.

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Tried cutting the coir in half with a butter knife, but it proved unweildy and I got an uneven cut. I then got out a bigger knife (the solution to many problems) and shaved until I had approximately half. I made a complete mess.

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The water was boiling so I killed the heat and threw in the coir. Since I chopped it up, it absorbed the water right away and I was able to immediately break it up and with a spoon to what appeared to be even moistening. I didn't wait to let it sit as it seemed to be well mixed. Maybe this was a mistake.

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The PH was 7 and wharfrat's thread mentions adding pickling lime to get to 7.5. This I believe is partly to correct for the coffee and 7 is acceptable, but I thought "what the hell, I'll try for 7.5". This is where things started to go wrong.

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After stirring for several minutes to get it even and waiting 20 minutes for the lime to set in I took the measurement again. Still 7! So what did I do? Biggest mistake of the night: added more lime.

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Uh oh...

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So with a PH so alkaline that my meter couldn't read it, I decided to dilute it by making the second half of the coir brick.

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In the meantime, I remeasured and got a different reading. Wondering if there was a clump where I was measuring before. Still though, it was too high. I reread a couple times and had similarly high results.

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All into one big pot it went! The PH balanced out wonderfully. Though part of me wonders if I didn't give it enough time to settle in its new home. I am now thoroughly convinced that using lime is magic.

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Time to ladle into my Reynolds large 8lb oven bags!

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I cut out little holes (aiming for two inches above substrate on either side, and one high up on either end). I then covered them in micropore tape.

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I put in something to raise the bags away from the water. I also put a plate on the top of the bags incase they expanded and risked blocking the vent of my PC.

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I then waited impatiently for the PC to come down to 0 PSI. Apparently bad things can happen if you force the pressure to drop to quickly.

 


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#12 PinkMenace

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Posted 08 December 2017 - 11:35 AM

Next I inoculated with cakes! I had 9 cakes, so I split them evenly with 3 for each bag.

 

Gathered my sanitary gear

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Birthed the cakes and rubbed off the verm barrier, lining them up before opening the bag.

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I started out grinding directly into the bag then switch to a bowl. After I was done getting the spawn in the bag, I mixed it evenly throughout the bag. The order of these photos is a little doctored.

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I then zip tied the bags at the very top to close them. Wrapping around a stick or bicycle spoke would have been preferable to keep the arch even/flat... but that's not what I did.

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After finishing all three bags, I tied them up in my myco shelving.

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The cakes were poorly made, with chunks of BRF that I don't think were able to be fully penetrated. I'm hoping for the best, but I have low expectations.

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One of the cakes had a sizeable pin, surprisingly.


Edited by PinkMenace, 08 December 2017 - 11:38 AM.

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#13 PinkMenace

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Posted 10 December 2017 - 09:47 PM

Update!

 

Did you know that when you split apart spawn, the resulting colonies can sense each other? I learned it in a book! Remember children, read and do drugs!

 

Approximately 67 hours from last photo of these bags (roughly 3 days): 

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And 7 days from making these jars (first signs of life were at day 6, yesterday):

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#14 coorsmikey

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Posted 10 December 2017 - 09:58 PM

Do you think the sense one another with conscience thoughts and language?

#15 CatsAndBats

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Posted 10 December 2017 - 10:05 PM


 

Did you know that when you split apart spawn, the resulting colonies can sense each other? I learned it in a book! Remember children, read and do drugs!

 

 

 

 

What book and how do they know? It's like when people say that dogs dream, they probably do but we don't know.


Edited by CatsAndBats, 10 December 2017 - 10:06 PM.


#16 425nm

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Posted 11 December 2017 - 02:44 AM

Fungi can certainly sense mechanical stimuli [1] as well as light [2,3], and flow [4]. I'm sure they can sense a variety nutrient gradients just like bacteria can [5]. Bacteria can even sense other bacteria via mechanisms such as quorum sensing which is often important in the formation of bio-films as well as triggering virulence responses (the way it works in Vibrio cholerae is quite interesting) [6]. I would be very surprised if they didn't have some mechanism for sensing con-specifics as well as other species be they fungi, or otherwise. I just can't dredge up papers on it quite so quickly as I don't know what I'm looking for. I suspect that [7] actually provides some specific answers but don't have time to read it tonight.

It often sounds crunchy to suggest that bacteria, or plants or other sessile organisms can sense things but if you stop to think about it of course they can. How else do they go about acquiring nutrients? or necessary minerals? Even sessile organisms have very active mechanisms for acquiring these things but first they have to know where those things are. Whether or not they have some collective experience of these senses, ie. a consciousness is a much harder question to answer.

As for whether or not dogs dream. I suspect that their brain waves should be similar enough that sticking an EEG on a dog would certainly provide some clarity but then you'd have to get an EEG on a dog. Surely someone has done this. Weirdly some sort of "sleep-like" behavior is seen in almost all life, at least to my knowledge. Everything seems to need to sort of power down for awhile. Its definitely one of the interesting edges of Science. All that being said Cat is definitely right that Humans are very good at seeing and believing what we want to. We're often hilariously wrong when it comes to animals.

[1] https://www.ncbi.nlm...pubmed/18679170
[2] https://www.ncbi.nlm...pubmed/17067849
[3] https://www.ncbi.nlm...pubmed/20451644
[4] http://www.sciencedi...168165697001478
[5] https://www.ncbi.nlm...cles/PMC426976/
[6] http://perspectivesi...11/a012427.full
[7]https://www.ncbi.nlm...les/PMC2998294/

Edited by 425nm, 11 December 2017 - 02:49 AM.

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#17 CatsAndBats

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Posted 11 December 2017 - 09:13 AM

And I'm a cat!

#18 PinkMenace

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Posted 11 December 2017 - 11:03 AM

The book was Organic Mushroom Farming and Mycoremediation by Tradd Cotter (the Mushroom Mountain person).


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#19 scott_1971_h

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Posted 12 December 2017 - 06:29 AM

I believe oyster mushrooms can also sense old nematode tracks, and send hyphae out to get a lovely bit of nitrogen.


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#20 scott_1971_h

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Posted 12 December 2017 - 04:21 PM

https://www.ctahr.ha...roomCompost.pdf

I'll be supplying my suburb oyster mushrooms, and next year Im expecting to supply it tomatoes. Do they work on snails?






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