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Mushrooms, Mycology and Psychedelics > Gourmet and Medicinal Mushrooms
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Agar problem
    #26032904 -

I don't know if this is the best place for this or not. Perhaps a moderator will be kind enough to put it in the right place if not. Thanks!

I started having a problem that I can't seem to figure it out.
This surely sounds like a newbie question but it concerns something I've only recently encountered despite not being a newbie.
When I pour plates they are all staying like a soup.  I've changed my agar to a new source three times now, tried an oatmeal/dextrose recipe instead of cooking potatoes for PDYA and have tried it with potato flakes (my most frequently used recipe being 22g potato flakes, 16g high-grade agar, 8 g dextrose and 1.5g yeast extract in 1L). 
I've also increased the agar from 16g/L to 24g/L and it made no difference.
I have now consistently failed five times in a row and feel rather lost.

I am using an AA925, with a 1L recipe split between two 1L polypropylene flasks (measured separately, not mixed and split), each one closed with a foam plug and a layer of tyvek tied over that, cooked for 45 min at 15 psi. 
The water is pH 6.74 well water with very little mineral content so it is a little on the acid side but it has been working fine for more than four years with completely consistent gelling.

Any suggestions of how to troubleshoot this are greatly appreciated. This only became a problem during May and before then plates came out fine. I am guessing it is something simple I am somehow not seeing.
Thanks!

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Re: Agar problem [Re: NPoB]
    #26032997 -

I've used 20g agar + 20g barley + 5g bacterial yeast + 1L water, after 45min to 1hour it will be solid jelly. T-Gel agar is also quality.

Pressure cook time is also 45min 15 psi.

It worked before consistently but not now?? Hate them riddles, something broken maybe?

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Re: Agar problem [Re: mojotoke]
    #26033005 -

You are not using enough agar.

My recipe is...

500 ml or 500 g Distilled Water
20 grams light malt extract
20 grams agar

For 1 liter you would need 40 grams agar.

The above recipe may work with 16 grams agar I never tried. I bet you were using 16 grams to 500 ml water and not 1000 ml water which is 1 liter.

Edited by SHROOMSISAY01 (06/04/19 06:36 PM)

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Re: Agar problem [Re: SHROOMSISAY01]
    #26033032 -

I also use more agar. My question,  do you pre-heat your agar before pressure cooking?

I will say my last pour had some wierd goopy plates. I wrote those off to possible bacteria.


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Re: Agar problem [Re: Quadman]
    #26033116 -

I use hot water to mix. Then I PC. When I take the agar out of the PC I swirl it around until it is mixed up. I use a polypropylene pitcher to do my agar. I have a flow hood so it is no problem for me.

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Re: Agar problem [Re: SHROOMSISAY01]
    #26033491 -

I also feel you should use more agar.

I go with 20 grams agar, 10 grams LME, 1-2 grams yeast and 1000 ml H20.

off topic but i like to use a lesser amount of LME to lessen bacterial growth.

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Re: Agar problem [Re: deadmandave]
    #26033996 -

Thanks all. 
I should have made it more clear. This has been the case even at 24g/L.
16g/L has been working great for me for four years (I've been using a high gelling strength agar derived from Gelidium) and, for the last two tries I've increased the agar to 24g/L and it is still staying like soup.
I've gone through almost three full cases of sleeves before having this problem (with 500 plates per case) so this is a really frustrating development.
I'm going to try 32g/L today but am growing weary of trashing so many plates to no avail. Plus there are transfers that are now way past when I wanted to do them so an important part of my production cycle has ground to a halt. This was working just fine for me until sometime in May and has not worked since.

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Re: Agar problem [Re: NPoB]
    #26034005 -

To answer the question, I do not preheat the agar.
I add dry powders to the flask and then add water, swirl it to mix, add a foam plug and Tyvek and cook.

If 32g/L does not work I will go to 48g/L but I am really baffled why this changed as it had been working fine.  You can see what I have been doing, including some nicely gelled plates, at naturalproductsofboonville.com

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Re: Agar problem [Re: NPoB]
    #26034032 -

Are you using an MEA or something like that? MEA has a mixture of agar in it? I don't think you are by what you said. But not mixing the dry MEA before you start could be a problem. I am sorry my friend I am baffled. But I would start with 40g/L and then You will know if it is not enough agar. Then each time decrease the agar a little each time until you end up with soup again. The agar has to be the problem.

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Re: Agar problem [Re: SHROOMSISAY01]
    #26034071 -

Thanks for the advice.
I don't like pre-made mixes as they can separate. Despite saying that, I do rarely use Stamet's pre-mixed antibiotic agar if the target seems to demand that but normally alternate between PDYA and ODYA with PDYA being used the most often. That is being done as 22g potato flakes (or sometimes as 200g fresh potatoes cooked in 1L water), 8g dextrose, 1.5g yeast extract and 1L water.
There are two flasks of PDYA cooking right now. One has 16g agar in 500 mL and the other 24g agar in 500mL. By this afternoon I hope to know more.

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Re: Agar problem [Re: NPoB]
    #26034157 -

What antibiotic is used to make antibiotic agar?
Cheers
gww

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Re: Agar problem [Re: gww]
    #26034246 -

I would have to look but I think Stamets might use gentamicin in that mix? It should say at the Fungiperfecti website. Most antibiotics are heat sensitive so need to be filtered at 0.22microns and added only after the agar starts to cool down.

I also think that I have solved my agar problem a few minutes ago and it was a *really* stupid mistake. I had repackaged some bulk materials into glass jars and apparently somehow had mislabeled a jar of citric acid as being organic non-gmo dextrose. That is definitely going to be causing a gelling problem. I was making up some liquid media in the event the plates still did not work for me today and on a hunch I tasted some of the "dextrose" so there is no question about my making that error. I've turned off what was cooking and will make up another lot today. Using the right ingredients this time. Which I anticipate will gel fine.

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Re: Agar problem [Re: NPoB]
    #26034435 -

Glad you found it. That can make you crazy:lol:


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Re: Agar problem [Re: Quadman]
    #26034901 -

It was certainly starting to drive me crazy. Everything gelled fine just now so the flow can resume. Thanks for the thoughts and suggestions.

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Re: Agar problem [Re: NPoB]
    #26035034 -

Well, I must be using some crap ass agar! What agar are you using for it to gell at 16 G/L?

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Re: Agar problem [Re: SHROOMSISAY01]
    #26035291 -

I've mostly been using "bacteriological grade" agar from Mycosupply for the last few years.  Don't confuse that with nutrient-containing media made for culturing bacteria though. This is a high gelling strength agar suitable for mycological or bacteriological work and contains only agar.
If I understand right, the best setting agars all come from Gelidium spp. (You can google "Gelidium agar" and find a wide range of suppliers.) The food oriented agars all make a softer set. Gracilaria gives a high grade agar but it sets softer. Lots of times agars get blended to reduce the price or sometimes to fulfill their supply needs. 
When I started, I was using 24g/L as that was what the recipe I had said. Another one said 18-22 so I tried less. It did not seem to suffer from being reduced to 16g/L. I was just curious how far it could be reduced. It is not as firm as with 24 but lets me do transfers just fine whether as small bits onto other agar or cutting it and moving the sections onto grain.

If your agar is taking 40g/L and it is working, it is still working. That is really the most important part and there is nothing wrong with it
How much it is taking per liter might balance against the price if it is less expensive?

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Re: Agar problem [Re: NPoB]
    #26035298 -

Your agar is old or old and oxidize..
All lab agar should have a expiration date to let you the shelf life.
Take a look It should be there.

Either use allot more or get another source..


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Re: Agar problem [Re: micelio]
    #26035355 -

Thanks. I shared your thought while I was still having problems.
It is working again now that I am really using dextrose instead of mislabeled citric acid. Duh.

It seemed worth sacrificing a plate to enable an image.
The image is of PDYA using 16g/L of agar. (as 8g/500mL) This was cooked and poured this afternoon using the same jar of agar as I had been using when the "trouble" began.



A triangle has been cut free to show how it gelled. More agar might give a crisper cut-line but this is working fine for me.

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Re: Agar problem [Re: NPoB]
    #26035981 -

NPoB
Thanks for answering my question earlier.
gww

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Re: Agar problem [Re: gww]
    #26053918 -

Not a problem. Happy to do so.

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