More Liquid Culture Discussion

Nan's Nook : Archives : Honey/Dextrose Q&A: Questions??? : More Liquid Culture Discussion
  Subtopic Posts Updated Creator
Maple Syrup?  4 10/11 05:47pm Brad
Honey tek / Mycelia growing in homemade syringes  6 11/30 01:48am Dr. Cubesis III
Question - liquid inoculation  17 12/20 06:12pm Ryan Waters
Quick HoneyTek/RiceTek inoculation Q  12 02/05 08:33am Martaxus
How could you clean Mycelium for honey tek inoculation  12 02/12 12:09am Pinhead

By fungis amongus (Fungisamongus) on Saturday, December 01, 2001 - 04:00 am:

I've recently started using a lof of mycelial water to grow shrooms. Why not? Its a lot easier for making large numbers of syringes, and they inoculate about 2 times faster than spores. So far the benefits kick ass.

But I have a potential problem and I need your advice.

I seem to remember once on the old board, that I read a post (by possibly Quote or Oldtimer) about mycelial water and a degenerative effect on the gene pool. (I may be wrong, thats why I'm asking) - But what I remember reading was that if you grow 1 generation of mushrooms from mycelial water you are fine, however if you take a spore print from that same generation and make another batch of mycelial water to grow a 2nd generation of shrooms, then you are going to get a degenerative effect of your 2nd generation and so on down the line...

I don't see any reason that this would be true, but I've been unable to find any more info. I posted once asking about it, but I was thinking it had something to do with a shrinking gene pool, and I was told thats not true.

I think it does have something to do with a shrinking gene pool. The question is: will this result in a downward spiral of potency? And if so, a suggested method to bring this back up might be to start again with a spore syringe. However, if you do that - you're still taking spores from a generation that has 'x' many less genes in the gene pool than the generation before it. (did that make sense?)

So in any case, whether I'm remembering right or wrong, I'd like any helpful info, clarrification, or experience you might have to offer. I would hate to continue this past my first generation if this is true. My whole gene pool could go to shit... (fuck thats the longest post I've ever written, I'm taking a bong hit now)

Thanks a million

By quote: (Quote) on Saturday, December 01, 2001 - 01:35 pm:

i believe that what i've warned against is using mycellial water itself to start additional mycellial water. you can only do that once or twice before returning to a fresh start from spores.

By ggg (Ggg) on Saturday, December 01, 2001 - 01:48 pm:

Cyrus Barnaby Tek

By Underground_Shaman (Shaman) on Saturday, December 01, 2001 - 06:22 pm:

This phenomenon you describe also sounds like cloning senescence problems. When you clone a clone through successive generations, you can have a senescence of the strain/cloned strain. This is due to a weakening of the "genes", but is probably related to shrinking telomeres.

However, if you take prints from your fruit, these spores are a-okay for starting liquid culture.

Good luck and happy shrooming!


By fungis amongus (Fungisamongus) on Saturday, December 01, 2001 - 08:52 pm:

I guess as a precaution, I'll just keep taking a spore syringe every other generation, and growing those mushies to print for a batch of mycelial water.

By fungis amongus (Fungisamongus) on Sunday, December 02, 2001 - 01:46 am:

Please keep the thoughts and input coming guys, I'd like to know any other experiences you may have had. Thanks.

By Mr. B (Argonaut) on Monday, December 03, 2001 - 04:28 pm:

Ok, on the Honey Tek, which btw I already started b4 I saw the Kayro syrup method. Here is what I did.

1. 1/2 pint Mason Jar.
2. 1 tbsp organic honey in jar.
3. added 100 ml boiled tap water.
4. Put lid on upside down w/ innoc hole in middle
screwed down snuggly with tin-foil on top
5. PC'd 30 min. (my PC has no gauge, new one
comming end of week)
6. let cool to room temp in PC. will not open PC
till room temp.
7. insert 2cc from syringe I made.
8. put in box in closet with ligh next to it.
It maintains about 73deg F.
9. Wait 3-5 days swirling it around daily.

First time with Honey tek would. Can I get a critique on my methods?


By quote: (Quote) on Monday, December 03, 2001 - 04:49 pm:

how is the inoculation hole sealed,
polyfil, taped ????
no real need to use 2 cc's of spores when .25 cc will suffice.
it'll go faster if you kick that temp up to about 80 or so.

By Mr. B (Argonaut) on Monday, December 03, 2001 - 04:52 pm:

Hole is not sealed. Just the tin-foil.. I see some folks here taping, some not taping, just using the foil.

By quote: (Quote) on Monday, December 03, 2001 - 10:14 pm:

well, mere foil isn't going to work, unsterile air will enter and contam it.
you need to plug that hole with polyfil, or tape it closed.

By Cragith Kilbonith (Kilborn) on Monday, December 03, 2001 - 11:53 pm:

you said 30 min pc, is that once its at 15 lbs of pressure or just 30 min? quote i think is right i think it will most likely contam unless polyfil is added.

By ion ewe (Ion) on Tuesday, December 04, 2001 - 06:17 am:

If your PC has a pressure regulator weight, you don't even need a new gauge unless it is leaking pressure during the cook. You see, the regulator weight is designed to always be correct. If you set it on at the 15 lb. hole, your pressure will be 15 lbs. + or - about 2 lbs. at all times. Unless you routinely scrub the crappin' hell out of your weight (making it uneven or lighter), all you need in the pressure gauge hole is something that plugs it; be that a good gauge, a broken one, or a stainless bolt. If you do PC with a busted gauge, make double damn sure your vent pipe is clean before each use! This is the pipe the weight sits on. Use a standard pipe cleaner on it.

My buddy has not yet determined what effect swirling has on the germinating of spores in a liquid inoculum. The theory is that they want to be still, for they can't very well germinate while still on the wind or in flux of any kind. Has anyone tested this? He still swirls because he can't help it!


P.S. Um... "Quo", I thought you may want to mention that if the polyfill gets wet or honeyed, it will wick contaminants right in from the air. My buddy uses gauze tape or "professinal grade" masking tape. Even for the beneficial oxygen and CO2 exchange, he can't get over the extreme probability of wetting the filter. Maybe a small bit of tubing cut so that it raises the filter out of the jar...

|*| * =polyfill
|*| | |=tubing
| |
------ <-lid
| |
______ <-level of sugar water

Just use a slightly larger bit for the hole in the lid. Just an idea... but it has been used successfully in other applications.

By An guy (Boomer) on Tuesday, December 04, 2001 - 06:34 am:

I swirl to expose the material to fresh nutrient.

By quote: (Quote) on Tuesday, December 04, 2001 - 11:59 am:

swirling also helps oxygenate the media.

By Cragith Kilbonith (Kilborn) on Tuesday, December 04, 2001 - 03:48 pm:

so............. 30 min at 15 lbs of pressure?

By quote: (Quote) on Tuesday, December 04, 2001 - 06:04 pm:


By Underground_Shaman (Shaman) on Thursday, October 18, 2001 - 06:34 pm:

In most renditions of this tek in the archives, the dextrose-water is sterilized with a Pressure Cooker. Does this imply that my old-fashioned stove-pot steam bath (used for BRF jars for years now) CANNOT be used to successfully sterilize dextrose-water? Would it work at all?

Do I need to convince my better half that we need a pressure cooker?

Second question, can a poor man's glovebox be successful for cloning and printing purposes? (My sterile technique is good) I'm thinking of using my old 10-gal. aquarium for this if it'll work. Any thoughts?

Looking foward to dextrose-water and cloning. I'm on a boogie-board riding the wave of the future



By Brettiejams (Brettiejams) on Thursday, October 18, 2001 - 07:56 pm:

Yes.... get a pressure cooker.

The regular old pot and with a lid just doesn't get hot enough to ensure propper sterilization with all sterile items.

They're really cool... you can even cook stuff super fast in them... a must have in my kitchen.

A well constructed poor mans glove-box can be fine for cloning or any other sterile work.

Although I am not personally familair with any desighns that use a 10 gallon aquarium, there is a good chance that someone else does

By Brad (Raze) on Thursday, October 18, 2001 - 08:30 pm:

You can make dextrose water with a microwave...

By Nan (Nanook) on Thursday, October 18, 2001 - 08:51 pm:

There is a bunch of places where you can cut corners on the Tek as we lay it out. But I don't want 50 people coming back and telling me all of their jars were contamed because their Honey/Dextrose was contamed. So we lay it out properly: providing detailed instructions and opinions on how to proceed. If you choose not to read up and/or not follow the Tek as we lay it out here, and you have problems, there can only be one place to put the blame.

By Joe Mamma (Madscientist) on Sunday, October 21, 2001 - 09:05 pm:

I have four jars that I have innoculated with mycelium from another BRF jar. these four are in the liquid form. I have innoculted a good number of BRF jars with this liquid innoculum and have had no growth for 13 days now. I am incubating at 90. I was told this was is a little high so I have dropped the temp to 85 fo three days still nothing. There seems to be no contaims in any jars. I have two jars with a little growth but these have seemed to stall. The growth in the liquid is large clouds of puffy white stuff not to much stringy mycilium going on. Is this normal. Two jars are three weeks old and two are about one week old. I am using honey and I know that the honey usually gets flakey. But this is not so flaky as it was in the begin this very dense puffy clouds. I have grown a number of times with great success but I have had bad luck for two months now. I have seem to beat my contaim problems but this is killing me. Any one have any ideas?

By Nan (Nanook) on Sunday, October 21, 2001 - 09:21 pm:

If you innoculated with mycelium... Was it chopped up or was it a chunk? Chunks don't work well for liquid culture. If mycelia is used to innoculate liquid substrate I make sure it is very finely chopped up. Macerated.

By Joe Mamma (Madscientist) on Sunday, October 21, 2001 - 09:32 pm:

I did not chop it up. It was small string like mycilium pieces. Is the clouds that I see mycilium or just the honey?

By Nan (Nanook) on Sunday, October 21, 2001 - 09:50 pm:

You are seeing mycelia I am sure. It just takes ages longers for the jars to grow out if you did not chop the tissue finely.

By Joe Mamma (Madscientist) on Sunday, October 21, 2001 - 09:58 pm:

Thanks for your input. Do you have any ideas why I seem to be having such a slow progression?

By Nan (Nanook) on Sunday, October 21, 2001 - 10:37 pm:

I think lack of chopping retarded your liquid culture, liquid culture needs mulitple innoculation points to perform well. Spores work well, macerated tissue works well (and is specified in several places). Dropping a chunk or a thread is not going to be nearly as effective as chopping up a much smaller sample and shaking to distribute.

Then (and this is a guess) you did not break up the mass of mycelium in the liquid culture jar before drawing up syringes... In other words the innoculum wasn't hot. You should be able to eyeball mycelial syringes and see floaters of mycelium everwhere. Use the needle to thrash at any clumps of mycelium in the liquid culture before drawing up.

By Mr. Tambourine Man (Tambourine_Man) on Friday, November 09, 2001 - 06:26 am:

Is room temperature OK for storing the Honey tek jars? I know it's not optimal, but will it work? Also, do the Honey tek jars need to be exposed to light? Thanks!

By quote: (Quote) on Friday, November 09, 2001 - 01:59 pm:

assuming room temp is within human comfort range,
it will work fine.
no need for light.

By Canadian Mushroom Grower (Canadian) on Thursday, January 10, 2002 - 09:57 pm:

I've been making honey water in the microwave and have had some problems. I'm going to make a glovebox and would like to know how to do the honey tek using a stove.I didn't see anything in the achives.

By Mr. B (Argonaut) on Thursday, January 10, 2002 - 10:06 pm:

Hey Canadian,

I have had some decent success with the mycellial water tek but I would recommend using the Light Karo Syrup since the untrained eye can spot any contamination more easily.

As far as using the oven tek to knock the jars up, that is how I did it and the only failures were due to opening the PC too soon and the jars sucked in "dirty" air with out my knowledge. As far as using the Microwave to sterlize the solution, I really couldn't tell ya how long it would take. If I PC them for 20-25 min I would think that would work for the Microwave also. 20 min in the nuke-o-matic is an awful long time. Maybe even 15 min would be ok. I'm sure someone that does it that way will respond soon enough.

Good Luck

By Dr. Cubesis III (Newbieshroomer) on Thursday, January 10, 2002 - 10:50 pm:

Hmmm, I just posted up a bit ago...

My friend bought some glass pyrex vials
and microwaved them
I posted his progress so far on todays threads..

By quote: (Quote) on Friday, January 11, 2002 - 12:45 am:

By Hudsonismss (Hudsonismss) on Thursday, January 31, 2002 - 07:16 pm:

1-can you use a small chunk of a colonized cake instead of fresh tissue or spores to inoculate the honey jar?
2-will the jar grow in temps around 50's-60's?
3-how long will the contense remain viable if left untouched after fully colonized (or whatever this growth is called) say in a freezer or fridge or shelf?

By ion ewe (Ion) on Thursday, January 31, 2002 - 07:27 pm:

1-Spores are used all the time. The tissue from a cake must be taken from the inside, just like regular cloning.

2-Very slowly if tissue is used. Possibly won't even germinate spores, though.

3-Do not freeze. They will die when all oxygen is used up. Refrigeration slows this process, but does not stop it entirely. It all depends on the original level of dissolved oxygen, the metabolism of that particular colony, and the "richness" of the liquid (the amount of food in ratio to oxygen).