Let's talk about Water Agar!! 💧

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Rotnpins
🤮 Rotten-Pins 🍄

Registered: 01/11/22
Posts: 4,738
Loc: in (front of) the hood
Last seen: 3 years, 7 months

Let's talk about Water Agar!! 💧

43
#27900393 - 08/12/22 10:03 AM (4 years, 8 days ago)

When I first started working with agar, I had an abundance of plates that I thought were too contaminated to clean up.. I thought about using antibiotics, but i had read that it could also hinder the mycelium growth and I didn't want to get stuck using it as a crutch.. So I decided to try making some water agar..

What is "water agar"?
Water agar is exactly what it sounds like, water and agar without any added nutrients.. the mycelium grows out very quickly (compared to the contams) looking for food.. this gives it a chance to out grow any contamination that may be riding along.

How to make water agar:
The recipe I use is 7g Agar powder per 500ml H2O

I leave the plates clear and add a different food coloring to each of my nutritional plates so I can easily distinguish between the type of plates.. (for example: H2O Agar=clear, MEA= blue, YMEA= yellow, etc,.)

You would prepare the water agar the same way as your nutritional agar plates.. the method I use is this:

1: add 100ml cold/room temp H20 to media bottle.
2: measure out remaining water (400ml for a 500ml batch) and heat water using preferred method. I personally bring it to a light boil on the stove.
3: while the water is heating, measure out your agar powder (7g per 500ml) and add it to your media bottles. Then give the bottle a little bit of a shake/stir.
4: once it is hot, add the remaining water to the media bottle. Put the lid on and shake it up.
5: prepare your PC with enough water for a 20 minute cycle.. put the water agar media bottle in the pc. If you have any nutritional agar, instruments that need sterilizing, no pour plates, etc,. you can throw them in the PC at the same time.
6: vent steam from PC for 10-15 minutes, add your weight, bring PC up to 15-17psi, reduce heat to low,, set a timer ⏲ and let cook for 20 minutes.
7: when PC cycle is done, turn the heat off and wait for pressure to drop to 0 on its own.
8: take your bottle(s) to your work space and wait for them to cool to pouring temp (I usually pour between 140-120 degrees)

How to use water agar:
Once your plates have cooled, it's time to do some transfers..

I make my transfers the same way I would when transferring to a nutrient plate.. I find an area that seems to have the most organized growth and I take a small transfer to the new plate.. smaller transfer wedges may be a little more difficult to work with at first, but they are less likely to carry contamination to the new plate. So try practicing taking smaller transfers each time until you're able to easily transfer a piece that is about the size of a grain of rice.

Wait for the mycelium to grow out.. it will be very thin and whispy, and it will grow out toward the edges much quicker than any contamination that tags along.

When the mycelium has grown out enough that I feel confident it has outgrown the contamination (its a good idea to let it at least half way to the edge, or all the way to the edge for more piece of mind), I take a few transfers from the water agar plate and move them to nutritional plates.

Next step is easy: watch your new plate as the mycelium grows out clean!

Edit:
Here's a video I found that discusses a couple applications for water agar that I didn't mention

Pictures added 8/28:

Here are some pictures to show you guys the growth on a few plates.. they aren't the best pictures, but you should be able to see the leading edge to get an idea of the speed of growth.

Plate stared 8/10 (18 days ago):

8/11 (17 days ago):

8/12 (16 days ago):

Agar to grain comparison
Water Agar vs Nutrient Agar

10/28:
I'm on a mission to start using up some of my ridiculously large plate collection.. So, I decided to do a side-by-side, plate to grain, comparison using 2 fully grown out Phobos plates.. One plate is water agar, the other is MYA. I decided to do 3 jars from each plate.

Quick picture before getting to work:

Here are the plates I'll be using

Plate 1 (MYA- cut into 3 wedges, leaving the center and outer ring behind:

Used plate 1 to inoculate 3 jars of oats:

Here is plate 2 (WA- cut in a similar pattern to plate 1)

Used plate 2 to inoculate the other 3 oat jars:

I plan to post updates comparing any differences in the length of time it takes for the mycelium to jump off, how long it takes the jars to colonize, etc...

11/3

The WA plates are finally starting to jump off to the grain a little bit.. they are definitely running a few days behind the jars that were inoculated using nutrient agar.. like I said in a previous post, this could be due to the age of the plates.. it is a possibility that the mycelium was in a dormant state, due to the fact that they're around 3 months old.. I still plan to run another test with fresh plates to see if the results differ from this batch.

Enigma (WA)

Phobos (WA)

Phobos (MYA)

Edited by Rotnpins (11/03/22 10:56 AM)

Extras:

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