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OfflineGrind365
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Registered: 08/07/22
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Awestruck] * 1
    #27902330 - 08/13/22 07:07 PM (1 year, 5 months ago)

Thanks Rotn.


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OfflineRotnpins
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Grind365]
    #27902339 - 08/13/22 07:19 PM (1 year, 5 months ago)

Quote:

Awestruck said:
That's a really cool, novel idea. Sounds good. Thanks for sharing the tek!





Quote:

Grind365 said:
Thanks Rotn.





No problem, glad you guys are finding the information useful/interesting :mushroom2:


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Invisiblebaz
Registered: 08/14/22
Posts: 3
Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Rotnpins]
    #27902741 - 08/14/22 06:42 AM (1 year, 5 months ago)

Can you germinate spores on water agar?


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OfflineRotnpins
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: baz]
    #27902778 - 08/14/22 07:19 AM (1 year, 5 months ago)

Quote:

baz said:
Can you germinate spores on water agar?




I haven't tried to do it, but I may toss some spores on a plate later and post the results ๐Ÿค” 

Personally, I like to use plates that have slightly less nutes and agar than my normal plates when making plates specifically for germination... using less agar will make the agar softer and there will be more water to help hydrate the spores.. using less nutrients will help the plate colonize faster, giving you quick growth to transfer to a new plate.

Sometimes you'll see spores germinate in a non nutritional syringe, so I think its possible the plate would germinate.. however I suspect that anything that does germinate will be thin and whispy and hard to distinguish what, if anything, would be worth transferring.

My recommendation for now would be to use nutrient plates for germination.. this will give the spores energy to germinate, and the mycelium will probably be easier to identify.. but I'll post my findings after I give it a try.


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Invisiblenosf3r4tu


Registered: 03/26/19
Posts: 775
Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Rotnpins] * 5
    #27902961 - 08/14/22 09:48 AM (1 year, 5 months ago)

I'm a lazy fuck and I just flip the bacterial plates and take a small sample from the other side. Myc grows through the agar, bacteria doesn't.


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OfflineRotnpins
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: nosf3r4tu]
    #27902965 - 08/14/22 09:50 AM (1 year, 5 months ago)

Quote:

nosf3r4tu said:
I'm a lazy fuck and I just flip the bacterial plates and take a small sample from the other side. Myc grows through the agar, bacteria doesn't.




Interesting.. I'll give that a try sometime and see how it turns out.. hadn't thought of that :super:


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Invisiblenosf3r4tu


Registered: 03/26/19
Posts: 775
Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Rotnpins] * 1
    #27902985 - 08/14/22 10:14 AM (1 year, 5 months ago)

Just don't push the scalpel through the other side. And take the sample from the middle of the plate (where the myc had time to grow through it).


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Invisiblebaz
Registered: 08/14/22
Posts: 3
Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Rotnpins] * 1
    #27905150 - 08/16/22 04:31 AM (1 year, 5 months ago)

I germinated tidal wave spores on water agar! It took 3 days to germinate and have many colonies while other plates itโ€™s not very visible. This pic quality isnโ€™t best but it is covered in myc colonies, white and wispy


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Offlinezilker
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Registered: 05/25/22
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: baz] * 3
    #27907494 - 08/17/22 11:25 PM (1 year, 5 months ago)

You beat me to it. 

I found an uncommon shroom in my back yard last weekend, Reddening Lepiota (red lip sounds better :cool:).  I left the spore print in open air until the ID was confirmed, then scraped into foil with a razor so it's contaminated to hell and back. Figure this is a pretty good study on the difference between the two agars?

Made 40 plates last night, half LMEY & half WA. Last task of the night was adding the spores to the plate. LMEY plate was cooled enough but had to wait on the WA until this morning...9 hour time difference.  Although I'm not sure what the myc is supposed to look like for this genus/species, I assume it doesn't resemble bacteria, as suspected in the LMEY pic.

Time difference...even after ~equal time, WA still looks less contaminated 9 hours later than the LMEY did.  I'm going on memory, didn't take pic, but I'll have time-equal pics soon and will edit post. Not sure either of these will make it, but hopefully it gives somewhat of a controlled study on how contam reacts to the WA??

Haven't USFSE, but wonder if anyone has tried spraying ISO on a spore plate with extreme bacteria contam, prior to myc growth?





8/18 - About 24hrs from last post, 48hrs from LMEY noc, and 36hrs from WA noc. LMEY is contaminated AF! WA is holding the contams back for the most part. WA has myc growth in two spots. Not visible in the pic and I barely caught it with a ray of sun today, but it's there.

The dark spot in the 1st microscope pic appears to be a mushroom crumb from the MS sample I saved.  2nd appears to be spore germination. 3rd is closeup of 2nd pic. Probably transfer WA to WA tomorrow.

Mind you, this mushroom was growing in the yard, finger fucked to death w/o gloves, and the spore print was left in open air to dry for well over 12hrs.  Pretty impressive IMO.

Again, nice job Rotn.

LMEY:


WA:


Microscope 1:


Microscope 2:


Microscope 3:


8/25 - Updated pics on the two plates.  Never attempted to transfer Plate #1 which is the nutrient plate b/c it never showed signs of growth.  Going in the trash today.  Made 3 transfers from Plate #2 on 8/19.  As you can see, although it did a great job keeping the bacteria at bay, it's now showing signs of the same mold (stachybotrys chartarum?) that #1 has.  I originally had plans to transfer to nutrient, but I think I'm going to transfer to water one more time to be safe.





I'll end the progress reports here and say that WA did the job it was supposed to do.  Took something dirty AF, cleaned it up nicely in 7 days, whereas it would have been trash with a standard nutrient plate.  Any future transfers are SOP and any future reporting would be redundant.  Cheers!


Edited by zilker (08/25/22 10:41 AM)


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Invisiblebaz
Registered: 08/14/22
Posts: 3
Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: zilker]
    #27909984 - 08/19/22 05:11 PM (1 year, 5 months ago)

Thatโ€™s awesome man! Great experiment and results. I will be using WA to germinate spores from now on and then transferring to MEA. Iโ€™ll post my transfer to MEA from WA when the myc starts growing


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InvisibleStipe-n CapMDiscord
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: baz] * 2
    #27909994 - 08/19/22 05:18 PM (1 year, 5 months ago)

Water agar is a great tool to have in the toolbox:thumbup:


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OfflineRotnpins
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: zilker]
    #27910001 - 08/19/22 05:23 PM (1 year, 5 months ago)

Awesome job man! Love the slide pictures to go along with it.. thanks for posting your journey :super:

I've had success taking a clean transfer back to MYA after just a few days if it's decently clean to start with... but if you can see contams tagging along, wait until the WA plate is almost grown out and then take a transfer from the leading edge just to be safe...

or grab a transfer or 2 right away, and hold on to the plate just incase they aren't clean.. by the time you see if the transfer was clean, the WA plate will be grown out a lot further from the contams


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OfflineRotnpins
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Rotnpins]
    #27910025 - 08/19/22 05:43 PM (1 year, 5 months ago)

Quote:

p9hu7 said:
Water agar is a great tool to have in the toolbox:thumbup:




For sure! I'm surprised how little it's been discussed around the forums..

I had heard it mentioned a couple times, but never with any results posted or certainty that it works..

I wish I had taken more pictures of some of my worst plates so I could share more of the results...

I just transfered some APE today that started out really bacterial and I thought it was a "trash plate". I didn't know about water agar when I was first working in this plate, so the mycelium grew through the bacteria and sat on this plate for a month before transferring to water agar.





This was the plate after transferring from water agar (I only left it on WA a few days, I didn't let it grow out very far, and it looks like it still did its job)



Note: I'm still learning, and haven't tested it on grain yet.. it still looks a bit disorganized, which is why I still did another transfer before doing a test jar.. I'll post the results once I have them


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OfflineJim I.T.I
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Registered: 07/01/22
Posts: 265
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Rotnpins] * 1
    #27913511 - 08/22/22 10:48 AM (1 year, 5 months ago)

thanks for this information, quite a bit in here i don't have experience with and great info added in from the community as well. next time i germ spores i will try on WA first

Cheers to all!


--------------------
Be patient & Let it happen


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InvisiblePsicomb
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Registered: 01/13/18
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Rotnpins] * 2
    #27913543 - 08/22/22 11:20 AM (1 year, 5 months ago)

Great write up, Rotn... appreciate the wisdom.

I'm doing my first water agar plates soon and then gonna transfer wedges from those into vials of sterilized distilled water for long term storage. 

I'm gonna use your recipe for sure :cool:


--------------------

When we constantly pull things apart trying to see how it works, we may end up with only an understanding of how to destroy something
- nick sand


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InvisibleBigwormS
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Psicomb] * 1
    #27913566 - 08/22/22 11:56 AM (1 year, 5 months ago)

Quote:

Psicomvb said:
Great write up, Rotn... appreciate the wisdom.

I'm doing my first water agar plates soon and then gonna transfer wedges from those into vials of sterilized distilled water for long term storage. 

I'm gonna use your recipe for sure :cool:



This is the first I'm hearing about this long term storage in steril water. Do you have a link to this tek? Or is as simple as, sterilize water in a jar and drop a culture wedge in. I thought I needed slants for long term storage.


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InvisiblePsicomb
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Bigworm] * 2
    #27913581 - 08/22/22 12:02 PM (1 year, 5 months ago)

So from what I understand certain species are a bit more finicky but cubes can handle it fine.

There isn't really a tek write up for it yet but there are bits of info from people who have had success from it here on the shroomery. 

I like the idea of water agar wedges into distilled watee because it is nutrient free, essentially putting the myc into a state of homeostasis...rather than having some super nutrient rich mix for the myc to keep eating, i.e. Slants. 

Plus ive heard it can be stored at room temp. 

This is all an experiment for me so I can't say what the results are.. but it's known to be an effective method for culture storage for various mushroom farms.

But back to the topic, water agar is specifically how to start that process :cheers:


--------------------

When we constantly pull things apart trying to see how it works, we may end up with only an understanding of how to destroy something
- nick sand


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OfflineHappyHigh
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Psicomb] * 1
    #27913590 - 08/22/22 12:08 PM (1 year, 5 months ago)

As I have no clean room can't even turn a/c off atm this was helpful.


--------------------
Had I the heavens' embroidered cloths,
Enwrought with golden and silver light,
The blue and the dim and the dark cloths
Of night and light and the half light,
I would spread the cloths under your feet:
But I, being poor, have only my dreams;
I have spread my dreams under your feet;
Tread softly because you tread on my dreams.


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Offlinerumfor69
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Psicomb] * 1
    #27913600 - 08/22/22 12:16 PM (1 year, 5 months ago)

Quote:

Psicomvb said:
transfer wedges from those into vials of sterilized distilled water for long term storage.




I always thought about doing this rather than making slants but wasn't sure how it'd work.


--------------------

Links For Beginners And Beyond


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OfflineRotnpins
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Re: Let's talk about Water Agar!! ๐Ÿ’ง [Re: Psicomb]
    #27913601 - 08/22/22 12:17 PM (1 year, 5 months ago)

That's a brilliant idea ๐Ÿ’ก  I was planning on making master slants of a couple cultures real soon.. I was going to do 2 slants of each (I always backup my backups :lol: ) but I think I'll probably end up doing 1 set of slants and one set of micro centrifuge tubes with water agar suspended in sterilized distilled water)..

I really like the idea of using the water agar for water suspension vs a wedge from a nutrient plate :super:


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