|
my_mycelium
Stranger

Registered: 09/04/22
Posts: 27
Last seen: 1 year, 2 months
|
Re: Let's talk about Water Agar!! 💧 [Re: Rusty2096]
#27934174 - 09/04/22 06:51 PM (1 year, 4 months ago) |
|
|
Oh absolutely! I apologise. I don't mean to hijack the thread!
Thanks!
|
Rotnpins
🤮 Rotten-Pins 🍄



Registered: 01/11/22
Posts: 4,738
Loc: in (front of) the hood
Last seen: 1 year, 1 month
|
Re: Let's talk about Water Agar!! 💧 [Re: Rotnpins] 3
#27935562 - 09/05/22 03:17 PM (1 year, 4 months ago) |
|
|
I started the experiment p9 and I were talking about

I may do a couple versions once I have some different things to work with. Such as putting bacteria in one corner and nutrients in the other, but for now I just did a very basic starting version to see what happens..
One sample is a transfer of Semperviva from a MYA plate and the other is a clean sample from the same plate. The hypothesis is that the mycelium will seek out the nutrient sample instead of just branching out evenly in all directions
|
Baba Yaga
♥ coir grower

Registered: 09/13/20
Posts: 3,955
Loc: Hyperspace Chicken Coop
|
Re: Let's talk about Water Agar!! 💧 [Re: Rotnpins] 2
#27935594 - 09/05/22 03:45 PM (1 year, 4 months ago) |
|
|
This makes sense in principal. I assume that the mycelium will direct growth towards the nutrient source due to nutrients leaching out from the MYA agar transfer. Makes me think that this will work best if you make a relatively soft water agar and up the nutrient strength in the piece of agar that is suppose to attract the mycelium. Maybe also experiment with preparing plates in advance and let them sit for a while to give the nutrients more time to leach. You could also try a dollop of BRF past on one side to lure the mycelium.
I feel like slime molds are more vigorous, mobile and directional when they are surveying their surrounding but there is no reason why other mycelium can't be directed when a gradient in nutrient concentration is present. Are there any published papers on the topic that do include a description of the experimental procedure with recipes?
Thanks for doing this. Will be interesting to see what happens.
|
Rusty2096
rah rah raw in Lady gaga



Registered: 08/23/22
Posts: 4,946
Loc: 🌌
Last seen: 3 days, 11 hours
|
Re: Let's talk about Water Agar!! 💧 [Re: Rotnpins]
#27935596 - 09/05/22 03:45 PM (1 year, 4 months ago) |
|
|
-------------------- Currently looking for nothing. You guys who sent me stuff are straight up awesome!. We don't own things - things own us. Semi-solid liquid culture (SSLC)
|
Rotnpins
🤮 Rotten-Pins 🍄



Registered: 01/11/22
Posts: 4,738
Loc: in (front of) the hood
Last seen: 1 year, 1 month
|
Re: Let's talk about Water Agar!! 💧 [Re: Baba Yaga]
#27935613 - 09/05/22 03:52 PM (1 year, 4 months ago) |
|
|
For sure.. that's a good idea to pre prep the plate and to mix up the nutrient and agar ratios..
Also, I took the Semperviva transfer from a clean plate.. it will be interesting to see what happens when taking a transfer from mycelium on a contaminated plate..
Hopefully this first attempt will give a decent baseline result to compare future versions with.
|
Rusty2096
rah rah raw in Lady gaga



Registered: 08/23/22
Posts: 4,946
Loc: 🌌
Last seen: 3 days, 11 hours
|
Re: Let's talk about Water Agar!! 💧 [Re: Rusty2096]
#27939486 - 09/07/22 08:56 PM (1 year, 4 months ago) |
|
|
Quote:
Rusty2096 said: WA vs BRF agar experiment
I'm posting here because I will be trying to clean a culture using water agar and I thought it would be fun as I will also compare the results with BRF agar. My logic here is WA could work due to the rapid expansion of the myc looking for nutes and maybe be able to outrun the bacteria. For BRF I was just thinking: if bacterias have a hard time in BRF cakes, maybe it will also have a hard time on BRF agar vs the original PDA plates.
In the picture below, PDA plates 1 and 4 were inoculated from grains coming from a jar noct with a very bacterial LC. As you can see it's still really bad.
I'm hoping the myc on plate 1 will manage top grow past the bacteria (towards where the purple arrow is pointing) to make a simple transfer. That being said I'm realistic and I'm not counting on it so much.
In case it doesn't grow past it, I made transfers from plate 4 to both plates 2 and 3. I used a scalpel to scrape a tiny amount of myc on the grain and then dipped my blade in 2 locations on both plates 2 and 3 (don't worry I did flame sterilize my blade between plates).

Anyways just a fun little experiment of how well it's gonna go to clean the original PDA plate transfering to BRF agar vs WA.
I will post my progress in a few day.

UPDATE
Not much growth yet from the 2 tiny tiny scraped myc transfers. But I do have some progress from another plate as per the picture below. The plate the transfers were taken from was PDA. My hypothesis is that the myc transferred to the BRF plate is still looking for PDA and the one transfered to the WA plate is going to expend looking for any nutes.
-------------------- Currently looking for nothing. You guys who sent me stuff are straight up awesome!. We don't own things - things own us. Semi-solid liquid culture (SSLC)
Edited by Rusty2096 (09/07/22 08:59 PM)
|
zilker
No Entitlements Here


Registered: 05/25/22
Posts: 64
Loc: Texas
Last seen: 14 days, 22 hours
|
Re: Let's talk about Water Agar!! 💧 [Re: Cob] 2
#27941546 - 09/09/22 06:48 AM (1 year, 4 months ago) |
|
|
Quote:
I wondered about germinating spores on water agar. You've had luck with it you say?
I'm thinking of making my next batch WA and trying some spores out on it. All my current plates are growing pretty slow at 8g LME. Thought about dialing that back a little next time I make nutritious agar plates.
Yes. When Rotn first posted this thread, I ran a study with a nutritional plate and agar plate...it should be on page 2 of this thread. The spores were collected from a backyard find, and never intended for growing. When I found out they're an edible genus, Reddening Lepiota, I wanted to grow it.
Dumped some spores on both plates and the contam that developed on the nutrient plate eventually led to me throwing it out within a few days. The WA plate showed signs of growth in 18hrs. Granted, I've since noticed the Lepiota is a super aggressive grower, much like Blue Oyster, so I wouldn't expect the same timelines from a cube. However, I think it's safe to say you'd get favorable results fairly quickly.
|
Rotnpins
🤮 Rotten-Pins 🍄



Registered: 01/11/22
Posts: 4,738
Loc: in (front of) the hood
Last seen: 1 year, 1 month
|
Re: Let's talk about Water Agar!! 💧 [Re: Rotnpins] 3
#27941574 - 09/09/22 07:22 AM (1 year, 4 months ago) |
|
|
Here's an update on the first nutrient sample test plate day 4:

So far, the growth is stretching out in all directions.. however I'm taking note of a few things that could be influencing the results...
1) the size of the mycelium transfer I took from the nutrient plate... I was working pretty quickly and since the donor plate was clean, i just took a normal sized transfer without giving it a second thought..
It looks like the mycelium is still feeding off the nutrients from the original plate.. the growth is still rather strong/aggressive.. It should be interesting to see what happens once it's grown out a little further, and its starting to run out of food.
2) The proximity of the nutrient sample.. Theoretically, the nutrient sample wouldn't need to be placed very far away from the transfer on the WA plate...
If our goal is to direct the mycelium in one direction, away from bacterial contamination, and then grab a clean transfer to a new plate; it would then make sense that we want the nutrient sample to be far enough that it wouldn't provide food for the bacteria, but close enough that we could use this strategy to grab a quick sample of clean growth.
3) As Baba Yaga pointed out.. the amount of time that the nutrient sample is on the WA plate before the test could play a role.. if the nutrients are allowed to start to leaching into the WA plate, it could be easier for the mycelium to detect trace amounts, while still remaining far enough to not effect the growth of any contamination that is present.
These are just a few of the thoughts/observations I have so far.. it's still pretty early to make any judgements about the outcome.
I plan to test out a few different distances for the nutrient sample in future tests. I'll also do some tests with smaller sized transfers, and try comparing plates that are prepared ahead of time vs ones that have the nutrient sample added at the time of the test..
Even if the mycelium continues to go out in every direction, the nutrient sample could make it easier to identify different types of mycelium growth when choosing where to take a sample when transferring back to your nutrient plate.
|
Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
|
Re: Let's talk about Water Agar!! 💧 [Re: Rotnpins] 1
#27941607 - 09/09/22 07:51 AM (1 year, 4 months ago) |
|
|
Sounds pretty good man, makes sense. I doubt that normal fungal mycelium will respond in the same way that slime mold does but it's still interesting to watch.
|
Rotnpins
🤮 Rotten-Pins 🍄



Registered: 01/11/22
Posts: 4,738
Loc: in (front of) the hood
Last seen: 1 year, 1 month
|
Re: Let's talk about Water Agar!! 💧 [Re: Stipe-n Cap]
#27941610 - 09/09/22 07:57 AM (1 year, 4 months ago) |
|
|
For sure.. even if the mycelium doesn't seek out the nutrients, I may continue to use this method for WA plates.. It should make it easier to choose a good spot to take a transfer from.. it's kinda hard to pick the best spot from a whispy plate.
|
Stipe-n Cap


Registered: 08/04/12
Posts: 7,623
Loc: Canada
|
Re: Let's talk about Water Agar!! 💧 [Re: Rotnpins]
#27941643 - 09/09/22 08:22 AM (1 year, 4 months ago) |
|
|
Yeah, either way it's still a great tool.
|
Smellyhobbit
Actual Retard



Registered: 04/01/22
Posts: 11,144
Loc: Stables
Last seen: 11 hours, 18 seconds
|
Re: Let's talk about Water Agar!! 💧 [Re: Rotnpins]
#27941648 - 09/09/22 08:24 AM (1 year, 4 months ago) |
|
|
Would there be any utility to putting the nutrients closer to hasten growth once it’s reached? Or is that fairly redundant.
|
SexBurrito
Wontons and WAP


Registered: 09/08/22
Posts: 322
Loc: United States
Last seen: 1 day, 11 hours
|
Re: Let's talk about Water Agar!! 💧 [Re: Smellyhobbit]
#27942135 - 09/09/22 03:22 PM (1 year, 4 months ago) |
|
|
Hey y'all first time posting but I've been lurking for years. Rotnpins i was thinking to test your theory could you do a swipe down the center of your wa plate with a contaminated swab much like a fencee then only leave a small gate for the healthy mycelium to travel through to the food source on the opposite side of the contamination "fence"?
--------------------
GYT with SexBurrito
|
CowsPoopShrooms
StillUphill


Registered: 06/08/22
Posts: 505
|
Re: Let's talk about Water Agar!! 💧 [Re: Rotnpins]
#27942882 - 09/10/22 01:31 AM (1 year, 4 months ago) |
|
|
Until nutrients become available at some point later in time.
Edited by CowsPoopShrooms (10/14/22 07:10 PM)
|
Rusty2096
rah rah raw in Lady gaga



Registered: 08/23/22
Posts: 4,946
Loc: 🌌
Last seen: 3 days, 11 hours
|
Re: Let's talk about Water Agar!! 💧 [Re: Rusty2096] 2
#27947344 - 09/12/22 07:17 PM (1 year, 4 months ago) |
|
|
*****EDIT: Results in red below so you that have to read all that again*****
Quote:
Rusty2096 said:
Quote:
Rusty2096 said: WA vs BRF agar experiment
I'm posting here because I will be trying to clean a culture using water agar and I thought it would be fun as I will also compare the results with BRF agar. My logic here is WA could work due to the rapid expansion of the myc looking for nutes and maybe be able to outrun the bacteria. For BRF I was just thinking: if bacterias have a hard time in BRF cakes, maybe it will also have a hard time on BRF agar vs the original PDA plates.
In the picture below, PDA plates 1 and 4 were inoculated from grains coming from a jar noct with a very bacterial LC. As you can see it's still really bad.
I'm hoping the myc on plate 1 will manage top grow past the bacteria (towards where the purple arrow is pointing) to make a simple transfer. That being said I'm realistic and I'm not counting on it so much.
In case it doesn't grow past it, I made transfers from plate 4 to both plates 2 and 3. I used a scalpel to scrape a tiny amount of myc on the grain and then dipped my blade in 2 locations on both plates 2 and 3 (don't worry I did flame sterilize my blade between plates).

Anyways just a fun little experiment of how well it's gonna go to clean the original PDA plate transfering to BRF agar vs WA.
I will post my progress in a few day.

UPDATE
Not much growth yet from the 2 tiny tiny scraped myc transfers. But I do have some progress from another plate as per the picture below. The plate the transfers were taken from was PDA. My hypothesis is that the myc transferred to the BRF plate is still looking for PDA and the one transfered to the WA plate is going to expend looking for any nutes.

EXPERIMENT RESULTS
Ok so here are my conclusions, and what I like to think I learned from this experiment (I may be wrong all the way, not sure).
Plates 1 and 2 were inoculated (2 inoc points each) with a VERY LITTLE AMOUNT of myc scraped off a dirty grain on september 4th.
Plates 3 and 4 were inoculated with normal size transfers from a very dirty plate on September 5th. Didn't expect those to work out at all. All my hopes were in plates 1 and 2.
This is what I see in the pictures:
- Picture 1: Slow but healthy looking growth and could be used for a transfer (sorry we can't see shit on that pic because of the BFR sediments I didn't handle properly).
- Picture 2: Only the left inoc point shows real growth. I think I scalpelled the microscopic quantity of myc too deep in the agar in the right inoc point. Myc looks a little weaker, I guess from the lack of sediments, but clean symmetrical growth. Could be used for a transfer.
- Picture 3: Solid healthy growth, somewhat asymmetrical (for now at least).
- Picture 4: The coolest of all. Look at that growth. The 2 arrows point some kind of "headhunting for food mycelium". Could definitely be used to take a transfer away from contam, if contam was visible. I bet those "food hunters" would outrun most tam. Could be used for transfer.
 
 
Conclusion: I think WA is definitely a great tool to have to clean cultures. I got lucky and I will proceed to transfers from plate 3. But if I did not get that lucky, I would have used the "headhunters" from plate 4 or even the think cool looking growth from plate 2.
I hope sharing this taught something to someone. At least it did taught me.
-------------------- Currently looking for nothing. You guys who sent me stuff are straight up awesome!. We don't own things - things own us. Semi-solid liquid culture (SSLC)
Edited by Rusty2096 (09/12/22 07:43 PM)
|
Rotnpins
🤮 Rotten-Pins 🍄



Registered: 01/11/22
Posts: 4,738
Loc: in (front of) the hood
Last seen: 1 year, 1 month
|
Re: Let's talk about Water Agar!! 💧 [Re: Rusty2096]
#27947489 - 09/12/22 08:35 PM (1 year, 4 months ago) |
|
|
Thanks for sharing glad to see some positive results..
Just an observation, plate 4- the strong growth on the right side.. I'm pretty sure that's from the way you placed the transfer.. the mycelium was pointing in that direction and was able to grow that far while still feeding off the original transfers nutrients.. I try and place my transfers with the mycelium facing down to encourage consistent growth from the start..
I'm just making assumptions, but personally I'd let the growth start to get more whispy before taking a transfer from that area, just incase something is feeding off those nutrients and riding along with that strong growth..
Water agar does move slower than nutrient plates, but the further you let it grow out from the transfer, the more likely you are to get a clean transfer.
From what I've seen, the growth through most of the plate tends to be whispy and very even, with the occasional area that looks a but more rhizomorphic.. the left transfer on plate 2 looks exactly like I'd expect it to
|
Rusty2096
rah rah raw in Lady gaga



Registered: 08/23/22
Posts: 4,946
Loc: 🌌
Last seen: 3 days, 11 hours
|
Re: Let's talk about Water Agar!! 💧 [Re: Rotnpins] 1
#27947520 - 09/12/22 09:02 PM (1 year, 4 months ago) |
|
|
Thank you for your observations. Very pertinent.
I'm confident I'll get many clean T3 cultures from these plates. But after reading your comments, I might give those plates a bit more time and not proceed with the transfers tomorrow.
Thanks for starting that WA thread, I gained some valuable experience and knowledge from it 
I also hope, in the future, this will help someone else clean a "hard to clean" culture. If it does help only 1 person, the time taken to share all this will be soooo worth it.
Edit: typos everywhere, my english suck
-------------------- Currently looking for nothing. You guys who sent me stuff are straight up awesome!. We don't own things - things own us. Semi-solid liquid culture (SSLC)
Edited by Rusty2096 (09/12/22 09:03 PM)
|
Rotnpins
🤮 Rotten-Pins 🍄



Registered: 01/11/22
Posts: 4,738
Loc: in (front of) the hood
Last seen: 1 year, 1 month
|
Re: Let's talk about Water Agar!! 💧 [Re: Rotnpins]
#27955926 - 09/18/22 11:38 AM (1 year, 4 months ago) |
|
|
Quick update picture from the nutrient sample experiment:

Growth is still even.. should be interesting to see if the growth gets stronger once it has a chance to feed off the nutrient sample
|
Rusty2096
rah rah raw in Lady gaga



Registered: 08/23/22
Posts: 4,946
Loc: 🌌
Last seen: 3 days, 11 hours
|
Re: Let's talk about Water Agar!! 💧 [Re: Rotnpins]
#27955933 - 09/18/22 11:47 AM (1 year, 4 months ago) |
|
|
I'm curious what would have happened if you did it this way:

I like to think the myc would have communicated that food is that way and growth would be around the nutes piece of agar (purely theoretical)
Edit: of course this would be a WA plate, forgot to write it on the drawing
-------------------- Currently looking for nothing. You guys who sent me stuff are straight up awesome!. We don't own things - things own us. Semi-solid liquid culture (SSLC)
Edited by Rusty2096 (09/18/22 11:48 AM)
|
Rotnpins
🤮 Rotten-Pins 🍄



Registered: 01/11/22
Posts: 4,738
Loc: in (front of) the hood
Last seen: 1 year, 1 month
|
Re: Let's talk about Water Agar!! 💧 [Re: Rusty2096]
#27956019 - 09/18/22 12:43 PM (1 year, 4 months ago) |
|
|
For sure.. I plan on placing the nutrient sample a little closer next time, but it still needs to be far enough that contams won't hitchhike to the new sample. Going to try it a few different ways to see what happens.
|
|