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Cultivating wild strains project


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#1 cujoloki

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Posted 17 November 2020 - 04:22 PM

Hi! Welcome to my landing pad for a most exciting new project I have undertaken. Recently I have been out on isolated hikes as an amateur mycologist. I have learned over the years various mycological skills from this forum. One of which is how to identify and expand upon already existing concepts and follow accordingly in the footsteps of those who have shared their knowledge.
This is nothing new to most of you guys I just feel inspired to share my poor man's progress into hopefully cultivating these wild finds on a grand enough scale to help them one day be more prolific in our area.
I enjoy cataloging the pictures I have taken and now have progressed into taking ziplocks with me whenever I go on a hike. Recently (a few months ago) I found a plethora of specimen and have been trying to isolate cultures of a definite laetiporus (unknown sub) and lions mane. Both found wild and cleanly stored in a Ziploc for dissection later.

First off I have pictures of the lions mane find and the chicken of the woods find.

Followed by the specimens in a clearer view. From here they were stored for 2 weeks in a fridge, until i could source a glovebox.

I will be working from scratch. Starting and progressing with rudimentary tools with a heavy measure if sterile procedure. Working in a non sterile environment. I fully intend and am working on making 100 kilos of spawn. I will be using a PC for all sterile media. So that's a plus.

After waiting for 2 weeks and finding a semi usable glovebox I proceeded to take many samples from the specimens on the 20th of October.

These being done on experimental rice flour agar I homemade were doomed to fail. And fail they did.

Thankfully I procured a host of specimens.

A week later having researched ketchup cups and ordered agar agar, I delved into my first agar adventure ever. Inspired by mycotopia itself. Pictures as following will tell the tale of my success.

I am well underway in this adventure and more content will be uploaded soon.

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Edited by cujoloki, 17 November 2020 - 04:25 PM.

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#2 coorsmikey

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Posted 18 November 2020 - 09:26 PM

Cool! I'm looking forward to seeing some updates here! Wishing you much success and possibly sharing some of those wild genetics.


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#3 cujoloki

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Posted 19 November 2020 - 10:49 PM

Thanks Mikey! I'll enjoy spreading the love when it come to that. After all I wouldn't know my ads from a hole in the ground in the mycology dept. If it wasn't for this place. The lions mane and chicken of the woods just happened to be peak specimen to be harvested.

I thought it possible to do this thread since the mycelium is clearing up very nicely on agar and liquid cultures are testing and passing very nicely. More on that later.

I thought I should post how I managed to get my agar game down without purchasing the luxurious Petri. My first month I used a substandard placeholder for a scalpel. It was a wooden carving knife I honed to a razor sharp edge. Even though I Pc'd it I kept seeing black pin mold on my transfer edges. This I replaced asap with a standard replaceable blade scalpel for under 10$. Included was a lot of blades!

I dived into agar with a potato water /cornsyrup /coffee solution. I know since its my first time I should be conservative but I always want to push the envelope and use exciting innovations. After some inspirational smoking and brainstorming I decide to add .25 grams of biofilm from fermented fish extract. Which is basically half fresh fish parts and half fresh cane sugar mixed homogenously and left to ferment. It's called FAA in Korean Natural farming.

Having made 250 gallons of biochar a few months ago I decided later to add some more exciting additives to the actual agar to increase the mycelium autoimmune response to voracious bacteria and fungi that are made in my biochar reactor. I digress.

My agar recipe for my first run follows.

- 450 ml of tap water boiled in 150 grams of finely diced potatoes
- 10 g agar agar
- 10 g corn syrup
- .25 grams of FAA biofilm
- 50 ml of your best coffee
- graduated measurment quart canning jar (good glass)

I started off with boiling the potatoes until they were fork tender. Use 500ml of water for evaporation. Strain off all water into a vessel then fill quart jar with 450ml of starch water. Weigh the agar on a scale. Toss it in. Weigh the syrup. Slide it in the jar. Weigh out biofilm. Scrape it into the jar. Seal it with a ring and lid. Shake it like it's hot. Using a nice thick clean kitchen towel grab the jar and Shake it for a good 3-5 minutes. You want this to be very mixed up. The constituents in the solution at this point must be completely dissolved before pressure cooking.

- PC @15 psi for 30 minutes

This gets transported back into the SAB @150 degrees Fahrenheit and poured into 50 ketchup cups. Gloves are used and alcohol is poured onto paper towels and rubbed all over EVERYTHING before anything is opened. My SAB IS cleaned before each use with a 10% bleach solution and later rubbed down with a healthy application of isopropyl.

Ketchup cups come packaged in a sterile bag. The ketchup cup bag is wiped down with 90% alcohol then left to sit in SAB until opening. I usually pour all 50 at once in 2 runs using the whole bag. I am a mad scientist with all this agar. I have 2 open air cultures for every room in my house just for curiosity sake. @[email protected] jokes aside I needed alot of agar to pull this off. This method proved to be cost effective and contaminated plates removed after 1 week was less than 5 percent. At 2 weeks one or two more would show a random contaminate. As a whole I feel it's very effective.

Edited by cujoloki, 19 November 2020 - 11:14 PM.


#4 rockyfungus

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Posted 23 November 2020 - 05:20 PM

This gets transported back into the SAB @150 degrees Fahrenheit and poured into 50 ketchup cups. Gloves are used and alcohol is poured onto paper towels and rubbed all over EVERYTHING before anything is opened. My SAB IS cleaned before each use with a 10% bleach solution and later rubbed down with a healthy application of isopropyl.

Ketchup cups come packaged in a sterile bag. The ketchup cup bag is wiped down with 90% alcohol then left to sit in SAB until opening. I usually pour all 50 at once in 2 runs using the whole bag. I am a mad scientist with all this agar. I have 2 open air cultures for every room in my house just for curiosity sake. @[email protected] jokes aside I needed alot of agar to pull this off. This method proved to be cost effective and contaminated plates removed after 1 week was less than 5 percent. At 2 weeks one or two more would show a random contaminate. As a whole I feel it's very effective.

Not sure if just with time we tend to get lazy, but I can't remember the last time I actually cleaned my SAB. I just spray in there with some soap water, wait a few minutes, wipe the top off to see, and go to work. Sterile procedures>sterile environment IMO.
 


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#5 cujoloki

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Posted 24 November 2020 - 12:57 PM

I like to clean my SAB everytime I can. The space I use is very unsterile. It's a miracle I can get anything isolated as it is. Although my open air test that's a week old has nothing growing in it.

Edited by cujoloki, 24 November 2020 - 12:58 PM.

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#6 cujoloki

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Posted 24 November 2020 - 05:46 PM

Well I thought I would post an update. First off I made some LC out of some nice transfers I ended up with. I have 1 lions mane and 1 laetiporus sulphereous at this point. The original agar media proved to be successful with the chicken of the woods and hericium.

I used the half pint jars with micropore tape for a barrier. I used a 2% Karo and water solution and added a splash of coffee to the mix. PC @15psi for 30 minutes. Let cool and transferred to sterile environ.

Using what looked to be great growth I inoculated the next day. I transferred, using a scalpel, 4 to 5 square centimeters of the mycelium of each of the species. Taking great care to select the best growth on the outside ring of the "dish". I flame the scalpel until glowing every time I transfer.

Steps are as follows.
First wipe down lc jar ,dish, and gloves with iso soaked towel.

Secondly, crack seal on the LC is followed by flaming the scalpel and popping the specimen dish barely open.
I either cool the scalpel in the agar or on a specified iso soaked towel.

Lastly, I slice the agar up. In this case I used about half a cup. After which I shovel the cut up pieces into the jar and secure the jar lid then the cup lid. The Lc is then shaken until the contents are mixed well.

Notes: these LC both failed due to my part. The chicken of the woods I tested a week later proved to be contaminated in all tests. The lions mane showed great results at first but I believe the lack of proper LC jar setups invited contamination into the LC.
No worries that's why I made plenty of backups on agar. The 1st lions mane syringe I made; I tested on plenty of agar. They all showed pleasant growth. I subsequently inoculated 3 5kg bags of sawdust/oat/biochar substrate made @ 80/20/+20grams respectively. These have proved to be slow growers and i regret not waiting later until the most vigorous reared its' head.

Notes on notes: I have since made the double tyvek and self healing ported jar lids that are comparable to Coorsmikey's LC jars that i came across since then. I am hopeful to transfer the very vigorous growth i have had on agar to these 4 LC's I have constructed. Also completed are 15 Pc's 7.2 kg mushroom grow bags. (More to come) As well as a 4x4x8 ft grow tent to flush those blocks. I have made a 3 head humidifier by hand with of fans as intake. (Also almost complete) Thanks for reading this and if you guys have any pointers feel free to chat.

Pictures included are what's left of the first run of 50 "dishes". Followed by a display of the 3rd transfers of the lions mane group and chicken of the woods group with dates. This picture quality could be better but I hope you enjoy.

Notes on mycelium growth:
I have observed that my laetiporus sulphereous and hericium both have very faint mycelium growth followed by wispy strands that form a thick mat after about a week. All the samples I have grown out have had some form of contam that showed up until now! I hope to see mature mycelium soon!

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#7 cujoloki

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Posted 26 November 2020 - 09:41 PM

Good news and ok news. The lions mane 30 ml LC have tested positive! (Pic 1) 2 days old. Thick mycelium growth in LC lions mane. (Pic 2)

7 contains spotted in Chicken of the woods transfers. (Pic 3,4) More were also made 2 days ago that seem well and the parent hasn't shown any bad stuff. (Pic 5,6) Made a 500 ml agar with a sawdust base using 100 grams of sawdust and potatoes 100g boiled for 20 min then strained and Karo 10g. I have a thing about the different agar colors. I think like the blue.

Will be knocking up sawdust bags soon!!

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#8 cujoloki

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Posted 26 November 2020 - 09:57 PM

On another note. I have some LC of reishi that wants to grow on top of the nutrient. This is wild. Normal? IDK.

It has thick growth at the bottom as well.

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#9 cujoloki

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Posted 28 November 2020 - 06:42 PM

Testing 4 cultures today. I test each syringe before I seem them worthy of a bag. Knocked up 5 myco bags with the 3 syringes that tested positive yesterday! They wil be stored in garbage bags in a 75-80 degree room for incubation and will be checked in a week. Let's hope they pan out.
Feeling Christmas yet? Here's a pic of my current load in the SAB. Everything on there is growing great with zero signs of contam. Chicken of the woods will be set to make a LC with in a weeks time!

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#10 cujoloki

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Posted 29 November 2020 - 09:08 PM

Ok. Making a LC out of this Chicken of the woods plate. All 8 transfers out of these 2 red parents have taken off on my white oak agar recipe. Everything is loving it.

Pic 1 and 2 are the one I chose to go in the LC and then the LC before I add it. I finally get to use the new lids I Made! I just hope its successful. I'll be adding 1 whole ketchup cup to the LC after I is everything and slice it the hell up.

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#11 cujoloki

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Posted 29 November 2020 - 09:09 PM

That plate was 6 days old. Transfer 5.

#12 cujoloki

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Posted 29 November 2020 - 10:00 PM

Here we go the Laetiporus Sulph. In a LC. Pretty excited to finally have this prolific culture.Should be ready to test In a weeks time.

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#13 cujoloki

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Posted 29 November 2020 - 10:10 PM

I'd like to show how the lions mane has progressed in 6 days. It shows very aggressive growth! This is my 4th transfers and their parent.

The parent looks like it may fruit?! (Pic 2) Picture 3 is the most aggressive t4 up close. It's so dam furry! =)

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#14 cujoloki

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Posted 29 November 2020 - 10:22 PM

Here are some jars of popcorn I inoculated with the old lions mane LC. 1 @ postshake (pic1). Picture 2 is all of them together. 3 are farther ahead than the rest. I think I will use these to g2g a few myco bags or fruit out in my grow tent on a rack. Haven't made the decision yet. I wonder if a 1x1 cedar shelf would be good for my grow tent... Thanks all!

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#15 jrh

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Posted 30 November 2020 - 06:37 AM

@cujoloki

 

What kind of ketchup cups and lids are you using? I have some little ziploc squares that don't melt @ 15 psi for 45 mins, but they're not completely transparent. Could you post a link?

 

Thanks.


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#16 cujoloki

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Posted 30 November 2020 - 08:59 AM

Jrh, on 30 Nov 2020 - 5:37AM, said,
@cujoloki

 

What kind of ketchup cups and lids are you using? I have some little ziploc squares that don't melt @ 15 psi for 45 mins, but they're not completely transparent. Could you post a link?

 

Thanks.

 

The cheap ones you get at Wal-Mart. 50 in a pack. Packed in 2 stacks lids on the left of transparent plastic sack; cups on the right.
The bag is sterile inside. Find one without a hole in the bag, get it home and wipe it down with isopropyl then put it in your Still air box.


Edited by cujoloki, 30 November 2020 - 09:19 AM.

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#17 cujoloki

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Posted 30 November 2020 - 09:04 AM

I may try to make this (the last pictures) lions mane a better isolate by carefully selecting the fastest edge on that t4 plate that isn't growing uniformly. All of these transfers were miniscule and after transfer four on this batch i'v yet to see uniform colonization.


Edited by cujoloki, 30 November 2020 - 09:17 AM.


#18 cujoloki

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Posted 30 November 2020 - 08:42 PM

I am fascinated by this lions mane LC test I made 7 days ago. Awesome looking! (Pic1) Some of the LC even splashed on the top and is growing. (Pic 2)

The last pic is all 3 tests. All lions mane LC. One born on the 26th the other 2 on the 23rd.

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Edited by cujoloki, 30 November 2020 - 08:43 PM.


#19 jrh

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Posted 01 December 2020 - 06:04 AM

The cheap ones you get at Wal-Mart. 50 in a pack. Packed in 2 stacks lids on the left of transparent plastic sack; cups on the right.

The bag is sterile inside. Find one without a hole in the bag, get it home and wipe it down with isopropyl then put it in your Still air box.

 

Thanks, so you're pouring agar like these were petri dishes. Nice find.

 

b9428ca4-f712-4ff2-a221-b8fa3a4ec6f1.e7b


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#20 cujoloki

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Posted 01 December 2020 - 11:53 AM

Yes one and the same. They're a godsend. Having 50 plates to choose from brought this project to where it is right now. So as of yesterday i took account of all i have going right now. I have 15 total Myco spawn bags colonizing right now. (all around 2.6kg of Sawdust/oat Substrate) Knocked up the last 7 last night. I only have 7 30cc LC knockers. I have in cold storage healthy cups of Lions Mane and Reishi and Cotw. I Have of each 750ml working culture and 3 more blanks that are waiting with the nice new lids. (thanks CoorsMikey). Also i have 2 half gallon popcorns with LM and 8 quarts (4 reishi and 4 LM). Ill be making more mycobags soon (Around 30 this time). I want to have Cotw in those and more for whatever i need. I wonder if this could be a business.. I just want to eat a shit tons of Lions mane and i think it will help my severely autistic adopted son medicinally. Butter seared lions mane fettucine here i come.


Edited by cujoloki, 01 December 2020 - 01:15 PM.

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