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Liquid culture question


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#1 Taproot

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Posted 06 January 2020 - 02:57 PM

For some reason my liquid cultures don't seem to get very large pieces of mycelium growing in them. It multiplies a bit over 7 days and then stalls. I've seen people's jars with mycelium that fills almost 75% of the volume of the liquid. Mine is about 25% at best.
I have used wedges of colonized agar and noticed they have multiplied faster than when starting an LC with a previous batch of LC.

What I use:
200ml glass beaker with a filter lid for gas exchange
4%-4.5% Sugar
Distilled water
Pressure cooked for 35 min @15psi
A heat pad that keeps the temp up to 78F - 81F

I swirl the jars 2 times a day and when the mycelium gets clumpy and wont break up I use the magnetic stirrer to macerate it.

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#2 RutgerHauer

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Posted 06 January 2020 - 03:05 PM

If you use multispore in a syrup only LC you don't have to expect it colonizing very much. 20-30% is when it is done and ready to use or get into cold storage. The conditions and pressure under water makes the mycelium grow differently compared to colonizing grain or other substrates, it will stay clumpy and small.

 

You might see better results when using agar wedges because the agar will get into the liquid and thicken the solution, the mycelium will grow better that way. I use a pinch of agar in every LC I make to have the mycelium to colonize more of the liquid, for this exact reason. It will almost fully colonize the liquid that way. 

 

Try 0.1 gram of agar-agar per 200 ml of LC. I am experimenting to find the right balance, I think this ratio will work well - 0.05 grams might be sufficient even.


Edited by RutgerHauer, 06 January 2020 - 03:23 PM.

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#3 Myc

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Posted 06 January 2020 - 03:23 PM

Do you use anything other than sugar and water?

 

Perhaps try adding some yeast extract as a nutrient source for more robust growth.

I also stir liquid culture on a stir plate but I'm not sure I'm getting what you're doing.

You said you "swirl the jar twice a day"

I place a magnetic stir bar into the LC prior to sterilization.

Inoculate and place on the mag stirrer apparatus. Let it stir 24/7 until the culture is thick enough to be ready.


Edited by Myc, 06 January 2020 - 03:28 PM.

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#4 RutgerHauer

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Posted 06 January 2020 - 03:34 PM

Swirling the jar once or twice a day for half a minute will work fine as well. I don't yet have a magnetic stirrer and always have had to go manual, no problem.


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#5 coorsmikey

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Posted 06 January 2020 - 03:51 PM

Try one off of the heat pad next time, it may take a lil longer but I suspect that it will live longer and better without the heat.


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#6 Taproot

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Posted 06 January 2020 - 04:24 PM

Fantastic advice everyone.

I'll try you advice and see if my results are better.

Thank you!
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#7 onediadem

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Posted 06 January 2020 - 08:36 PM

https://mycotopia.ne...html?1037240816


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#8 sandman

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Posted 08 January 2020 - 11:26 AM

edit: whoops I see that you used light corn syurup not sugar hah.

 

4.5% is too high try 2%.


Edited by sandman, 08 January 2020 - 11:28 AM.

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#9 sandman

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Posted 08 January 2020 - 03:50 PM

https://imgur.com/TB0Cld8

 

man I'm having a total dickens of a time sharing a video in either mp4 or webm format, the forum seems to be blocking me from using the bb code or html code to share the video links for some reason. But here is a imgur link showing some lc action.


Edited by sandman, 08 January 2020 - 03:50 PM.

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#10 onediadem

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Posted 08 January 2020 - 07:02 PM

That LC good? It looks pretty tragic.



#11 sandman

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Posted 09 January 2020 - 07:46 AM

It's 100% good, the malt always stains the glass above the water level but no issue.



#12 sandman

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Posted 09 January 2020 - 08:09 AM

Heres a little better lit current stir shot

https://imgur.com/a/rWUbNnh


Edited by sandman, 09 January 2020 - 08:10 AM.

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#13 RutgerHauer

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Posted 09 January 2020 - 08:47 AM

That LC is lit.


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#14 onediadem

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Posted 09 January 2020 - 11:04 AM

That is crazy @sandman..  Nice lc in the second pic.


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#15 CatsAndBats

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Posted 09 January 2020 - 11:26 PM

https://imgur.com/TB0Cld8

 

man I'm having a total dickens of a time sharing a video in either mp4 or webm format, the forum seems to be blocking me from using the bb code or html code to share the video links for some reason. But here is a imgur link showing some lc action.

I just do quick gifs.

 

OP, regarding your percentages, I would absolutely follow @sandy's advice, but with a caveat, I'd also try some other percentages, but include a drop of honey, a drop of molasses, use brown sugar, etc (brown nutes), which include micro-nutes which can go a long way in helping myc to excel IME.

 

Here's aa+ on 4% nutrition (I forget which one/s, prolly sucrose and a "brown" nute)

 

 

post-147940-0-71175200-1578629835.gif

 

 

Another tip that I picked up, in lieu of a magnetic stirrer, use a piece of broken glass, it shreds the myc up like a motherfucker, akin to shaking a grain jar, pro tip, use colored glass. Props to @microbe for turning me on to the broken glass tip.

 

I also recommend that one doesn't germinate spores in LC, too risky IMHO. I use agar plugs.

 

 

 

 

 

 

 

 

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#16 Taproot

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Posted 28 January 2020 - 01:40 PM

I took the advice about using malt instead of corn syrup and I have a follow up question.

After I pc'ed about 13.7g of dry light malt extract with 460g of distilled water I was left with a heck if a lot of insiluable sediment in the jar. Should this be there? Maybe I should have boiled the malt first then filtered this out before running the pc?

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#17 sandman

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Posted 29 January 2020 - 09:37 AM

3% is too much malt Taproot. Try 1 - 2% or even less. I don't even weigh it anymore I just put in a standard spoons worth to 500ml, like the kind you eat with. Probably about 5-6 grams. There will be a little sediment no matter what though.


Edited by sandman, 29 January 2020 - 09:38 AM.

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#18 Fungi2b

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Posted 02 February 2020 - 02:34 PM

I took the advice about using malt instead of corn syrup and I have a follow up question.
After I pc'ed about 13.7g of dry light malt extract with 460g of distilled water I was left with a heck if a lot of insiluable sediment in the jar. Should this be there? Maybe I should have boiled the malt first then filtered this out before running the pc?


Leave it in there I find it seems to help, I think it gives the myc something to attach and grow on.

#19 Taproot

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Posted 09 February 2020 - 11:16 AM

Liquid culture successful! Thanks for all the suggestions


It took 5 days to grow this large.
First picture at 3 days, second at 5 days.
Temp of 78-80 F.
Innoculated with a 1cm x 1cm chunk of Albino penis envy on agar

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Edited by Taproot, 09 February 2020 - 11:18 AM.

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#20 stmhunter

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Posted 09 February 2020 - 01:10 PM

i put a couple marbles in the jar.






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