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Offlineoneirobotix
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Registered: 03/12/02
Posts: 12
Loc: presently: earth
Last seen: 14 years, 5 months
Liquid Mycelium Emergency Surgery
    #577096 - 03/12/02 06:30 PM (14 years, 8 months ago)

Hi all...

I admittedly have been lurking here for years, and I have waited until now to speak.

I finally decided to try the liquid mycelium technique (light malt), and I made an error in my calculations. I don't have a scale, and screwed up converting weight back to volume.

(Pause for laughter)

Okay, so here's my question. If I'm not mistaken I should use ~1ml dry malt to every 100ml of water. I currently have a 1:5 ratio that has been inoculated. (It has the color of newcastle brown ale, for reference). I don't know if anyone has done this before, but advice from some of the people who have been using the liquid tech. would be most appreciated....

The plan:
My spores have been sitting in the malt rich solution for 4 days.
1.Are they still viable?
2.Could I innoculate a new malt jar with the old solution?

Thank you for any assistance, and glad to finally make your aquaintances.


--------------------
life is but a dream...


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OfflineAzure
old hand
Registered: 01/01/99
Posts: 469
Loc: California, USA
Last seen: 14 years, 7 months
Re: Liquid Mycelium Emergency Surgery [Re: oneirobotix]
    #577148 - 03/12/02 07:15 PM (14 years, 8 months ago)

1.As long as there is no contamination in the solution and the spores were viable to start off with, you should be all good. You definitely have a little bit too much malt in the water, but it should not stop germination. The main problem you may have is sucking the mycelium up into a syringe-it gets really hard when you use too much malt. I don't know the exact concentration that I use, but I always eye ball it and it always works out just fine. I mix malt with water until the water is slightly piss yellow. The mycelium will grow just fine with this concentration and is easy to suck up into a syringe.
2. I don't recommend innoculating another jar with the spore solution because you'll increase the risk of contamination. Just wait for the spores to germinate and produce the mycelial solution. Make sure to keep the solution at 78-86 degrees F. Once it's ready, suck it up in a syringe, innoculate your jars and save a little to innoculate a fresh liquid solution as backup.


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Offlineoneirobotix
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Registered: 03/12/02
Posts: 12
Loc: presently: earth
Last seen: 14 years, 5 months
1000 Thank yous! [Re: Azure]
    #577184 - 03/12/02 07:37 PM (14 years, 8 months ago)

Azure,

Thank you so much. All I wanted was someone to calm my fears about this. Since this is my first trip out with a non-spore tech, but I just got tired of waiting for population of my jars as they dried out....and ultimately cut my overall output.

I stumbled across a pic this morning here, and started banging my head when I saw how light it was.

One more question:

Do you think that this will effect mycelium growth? (i.e.- slow it down)

Again.... Your reaffirmation is greatly appreciated!

Thanks


--------------------
life is but a dream...


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Offlinecookiewhore
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Registered: 01/03/02
Posts: 385
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Re: Liquid Mycelium Emergency Surgery [Re: oneirobotix]
    #577493 - 03/13/02 01:36 AM (14 years, 8 months ago)

at least you have good taste in beer :smile:

Hey, i have had ZERO luck with liquid cultures.

what did you inoculate the 4% ME water with?  spores? grain? agar?

ive tried agar and spores (B+) with no luck
perhaps another strain would work?


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Offlineoneirobotix
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Re: Liquid Mycelium Emergency Surgery [Re: cookiewhore]
    #577584 - 03/13/02 03:57 AM (14 years, 8 months ago)

cookie...

I used a spore syringe from a print that I made about 2 years ago. (I had to lay off farmin' for a while due to living situations.) Anyway, I made the syringe and let the spores rehydrate for about 36 hours.

I'm not sure if this will even work due to the overly rich malt extract, but I'm gonna hold out for a few days based on Azures comment. I was just gonna start over; experiment with my current solutiond; and make a new print.

I luckily have another print. It's mexicana if I remember correctly. (as this one was)

The liquid thing is new for me, but it makes a hell of a lot of sense. My jars always seemed to take forever coming straight from spores. I'm hoping this will speed up the process. I think that I'm also going to just go for mycelium instead of trying to mess with fruit. A friend of my recently told me that he has had great success using rye and "rinsing" out the mycelium with several OJ baths. I'm doing this for me and not to transport... so that's good enough.

If you'd like I'll keep you posted.


--------------------
life is but a dream...


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Offlinekarlfinn
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Registered: 03/10/02
Posts: 162
Loc: Wild, Wonderfu,l West Vir...
Last seen: 14 years, 4 months
Re: Liquid Mycelium Emergency Surgery [Re: oneirobotix]
    #577664 - 03/13/02 08:45 AM (14 years, 8 months ago)

hi...never know if / when a thread has ended....anyway...
showing good myc. growth, i would use it to re-innoc. a lighter jar.
sterile marinade injectors make for easy myc. sucking.
some chips of broken glass in the jar to start..shake...suck..squirt..
honey tek has woked fine...one tbs / pint..but switching to light malts and dextrose.
all broths for myc. growth are basically agar recipes sans the agar...
submerged culture tek is the fancy name.....applies to all that fermets..
I am pretty new to the liquid methods also...but as long as it grows..
and is kept xcontam ...its pretty flexible......lotzzz of recipes out there
and always...R & D......care...


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Offlineoneirobotix
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Re: Liquid Mycelium Emergency Surgery [Re: karlfinn]
    #577940 - 03/13/02 04:34 PM (14 years, 8 months ago)

karlfinn

a thread is never dead as long as valuable info can be shared. Due to the dark nature of my substrate right now, I'm unable to tell if the floating whispy things I am seeing are mycelium or just carmelized malt adhering to itself.

Your broken glass suggestion will definitely be taken to heart, as I feel itwould help prevent this.

Here's a queston for you:
I have to be relatively careful when agitating so as not to squirt the medium through the innoculation port. The lids are all covered with aluminum foil that p-cooked them with. My question is: with the glass being so heavy, how do you agitate it? Are you using another method for gas exchange? If I were to apply a hepa filter to the lid, are they still functional if the media gets them wet?

Jesus, I'm long winded...
Summary:
How do you agitate with glass without losing medium?

Thanks again for your input.


--------------------
life is but a dream...


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OfflineAzure
old hand
Registered: 01/01/99
Posts: 469
Loc: California, USA
Last seen: 14 years, 7 months
Re: Liquid Mycelium Emergency Surgery [Re: oneirobotix]
    #578270 - 03/14/02 12:28 AM (14 years, 8 months ago)

If you're talking about psilocybe mexicana, I don't know if those spores will germinate. I observed a print that germinated just fine when it was a few months old. However, a year later, it was observed that the spores would not germinate on agar. Three sucessive trials with different concentrations of malt were tried with no sucess. Either the print was contaminated or the P. mexicana spores have a short shelf life.
If you're talking about Psilocybe cubensis from mexico, then I doubt the strong concentration of malt will hinder germination.
The way you can tell if your liquid culture is bacterially contaminated is by checking to see if the liquid is cloudy. This can be very confusing because the mycelium will look like cloudy jelly. However, if the liquid ITSELF and not the chunky jelly is cloudy, then it is definitely bacterialized.
On agar, a high concentration of malt caused more rhizomorphic growth and did not slow it down. In liquid cultures, I have noticed that higher concetrations speed up growth, but I don't know about extremely concentrated soltions.


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Offlineoneirobotix
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Phew... they aren't mexicana! [Re: Azure]
    #578325 - 03/14/02 01:57 AM (14 years, 8 months ago)

I stand corrected. I looked back to the place that I got my original spore print from, and it is indeed PC. Yay! I had 4 jars of solution so I figured, what the hell. I made 2 new jars with a lesser concentration, and infused them with liquid from one of the other jars (My own little experiment) I have noticed significant clumping in the darker jars (high malt concentration), but at this point I can't ascertain whether or not this is caramelized malt or actual growth. (Tomorrow marks 1 week.) I just did the lighter jars, so I won't know for a few days on those, but well see. Hell, if it all goes down the pipes I have another print to go from. Again I thank you for your wisdom. This end of things (liquid) is quite new to me. I'm hopeful though.... and quite excited. It's been a while since I've done the DIY.

Thanks again

bluest wishes.


--------------------
life is but a dream...


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Offlinekarlfinn
member
Registered: 03/10/02
Posts: 162
Loc: Wild, Wonderfu,l West Vir...
Last seen: 14 years, 4 months
Re: Liquid Mycelium Emergency Surgery [Re: oneirobotix]
    #578495 - 03/14/02 07:37 AM (14 years, 8 months ago)

I use quart jars and corona beer bottles, clear and narrow necked, old-school, with poly-plugs..some gallon jugs with the same poly-cap
but the size is a pain to sterize even in my 941 cooker...so...
swirl around changing directions....don't shake and no, filter disks do not like
"wet" . In the jars...I have one small hole for injections and another larger 3/8 th inch, for a poly-plug.
wirks fine except the poly will wet-contam if not maintained...
I place several small and sharp glass pieces into jars before sterization, of course,
I an setting up a more pro submerged culture system...using an air pump and spone to agitate, 5 and 15 gallon containers,
caution, filter the air-pump air....or. yuk......also best to get myc. going with other mycelium innoculation, not spores..
Adam Gottlieb, free on the net, growing 5000 doses or something, has good liquid info, see Stamets..GGMM



and check out the gourmet and medicinal forum as these folks do this ALOT.....
P C .jars : 12-15 lbs / 30 min. do not exceed 15 lb or carmel....and less food for the beastie..


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Offlineoneirobotix
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Re: Liquid Mycelium Emergency Surgery [Re: karlfinn]
    #579245 - 03/15/02 01:28 AM (14 years, 8 months ago)

Gotcha on the pcooking... no worries there.

I've posted some pics below. Maybe (since you have a better idea as to what you're looking for) you can tell what I have going on here. I can say that whatever it is it's getting bigger. At first I thought it as malt falling out of solution but now I can't tell. It's really "wispy" after agitation. I'm intrigued by the air pump agitation. Nice idea. Let me know how it pans out.


before agitation

and after agitation.



Whaddya think?

cheers.

bluest wishes and thanks again.





--------------------
life is but a dream...


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Offlinekarlfinn
member
Registered: 03/10/02
Posts: 162
Loc: Wild, Wonderfu,l West Vir...
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Re: Liquid Mycelium Emergency Surgery [Re: oneirobotix]
    #579407 - 03/15/02 07:18 AM (14 years, 8 months ago)

hey pics are worth a whole lotta explaining...something is growing , looks loike myc.
thing is it IS growing so must not be too bad in there. no obvious contams ?? smells..?? I shake about two times a day
swirl until a little tornado forms then reverse.....
without the aid of other mechanical agitations. The air pump is actually a
commercial and lab mycelia production tek...see www.the forestfloor.org
under advanced cultivation. forum also www.mushworld.com
many medicinals are only from the mycelia......fermentations....
what are you planning on doing woth it ...next step....
I'd still try to suck out part of that and start several new jars with it...
see what happens with other broth mixtures...I am running puerto rican liquids right now
and they are so fast and hungry...


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OfflineBeppoMarx
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Registered: 09/01/01
Posts: 1,126
Last seen: 11 years, 8 months
Re: Liquid Mycelium Emergency Surgery [Re: Azure]
    #579628 - 03/15/02 01:24 PM (14 years, 8 months ago)

i measure 1:15 corn syrup to water in ML. i guess on teh jar by just filling 1/2 pint to the 100ml line or; if any a lil over. then removing the needle from the syringe makes it easier to suck up the thick mixture and i measure it with the syringe.


--------------------
Holy shit people; COMMON SENSE! we were all born with it where did it go?
maybe theres a tek out there to explain how to use it!
BUCKETS BRIGADE left hand man!!!


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Offlinekarlfinn
member
Registered: 03/10/02
Posts: 162
Loc: Wild, Wonderfu,l West Vir...
Last seen: 14 years, 4 months
Re: Liquid Mycelium Emergency Surgery [Re: karlfinn]
    #579726 - 03/15/02 03:41 PM (14 years, 8 months ago)

never posted a pic before so here goes,ahhhhhhhh,,,
these are a few of a puerto rican syringe.. in honey tek, @ 5 days @ 85 F.
two diff. jars, running close....my cambo is slower...and plantasia slowest of this test...
neither shown...
pref. of brew...setting up to make my own stout...sooo, much cheaper
mountain spring water, mmmmmmmmmmmmm, oh, yes i'm back.......
and I know all those fermeters are out there doing the very same thing...luck
oh, I have switched to dextrose powder from brew suppliers/shops. not corn syrup but corn sugar...
have a recipe r & d coming together with six differing broths...all with isolated strains running.
maybe some pics later, gotta get a better camera for closer shots...care....farmer....



hope I got this right, let me know if you see anything...
this is one tablespoon honey to one pint, nothing else, except tlc...


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Offlineoneirobotix
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Registered: 03/12/02
Posts: 12
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Re: Liquid Mycelium Emergency Surgery [Re: karlfinn]
    #580062 - 03/15/02 10:54 PM (14 years, 8 months ago)

man.... you rock harder than elvis.

I'm so excited. The new jars (from the transfusion) have already confirmed my thoughts. They're much clearer, and I have little fluffy clouds going on. Yes!!

Thing is I have 6 of these bad boys now.

Can you say bumper crop?

Being a farmer I'm sure you can.....

on another note... Please do keep me informed on the R&D. Sounds mighty interesting. I'm gonna try a few different substrates for further myc growing.

I really appreciate everyone's input here, and I thank you karlfinn for the pics and continued input. Glad someone else is knockin' the ol' grey matter around to try new things.

cheers....

oh...the song in my head tonight: Little fluffy clouds.

bluest wishes.
o


--------------------
life is but a dream...


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Offlinekarlfinn
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Registered: 03/10/02
Posts: 162
Loc: Wild, Wonderfu,l West Vir...
Last seen: 14 years, 4 months
Re: Liquid Mycelium Emergency Surgery [Re: oneirobotix]
    #580349 - 03/16/02 07:05 AM (14 years, 8 months ago)

peace and will keep u posted , at least the good stuff....farmer..


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Anonymous

Re: Liquid Mycelium Emergency Surgery [Re: karlfinn]
    #580354 - 03/16/02 07:17 AM (14 years, 8 months ago)

those are looking good Karlfinn..
Be sure to post some more pics in the comming days.

will you be doing a methanol extract on that?


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Offlinekarlfinn
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Registered: 03/10/02
Posts: 162
Loc: Wild, Wonderfu,l West Vir...
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Re: Liquid Mycelium Emergency Surgery [Re: ]
    #580571 - 03/16/02 02:33 PM (14 years, 8 months ago)

knock on wood..
I have not done alc. extracts before, but for now, no just cool dry the myc. not this one,
probably like to work with ethanol if that's the case.....dual usage....
this is just a multi-spore, I want to finally dry a more selected sub-strain, ...
but for now this one has eatten it's food and time to move on.
plucking a strong bit of this out to start another jar......actually, a few more,
change the broth recipe once in a while to keep 'em running true longer...
you know like agar recipes....and all else, we all get bored eventually..
later......................pics will follow along my r & d results , as I can.....


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Anonymous

Re: Liquid Mycelium Emergency Surgery [Re: karlfinn]
    #580776 - 03/16/02 07:59 PM (14 years, 8 months ago)

are you using a saccharimeter on this project?

Ive been looking long and hard for one w/ no luck..

Anyone know of online suppliers?


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InvisibleNighted

Registered: 11/10/01
Posts: 564
Loc: U.S.A.
Re: Liquid Mycelium Emergency Surgery [Re: oneirobotix]
    #580878 - 03/16/02 10:45 PM (14 years, 8 months ago)

Try 1 tbls Malt & 1 tbls Dextrose per 250ml of water. Works great for clones.



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

Freedom defined is freedom denied.


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Amazon Shop for: ½ Pint Jars, Agar, Air Pump, HEPA Filter, Malt Extract, Paul Stamets, Pressure Cooker, Rye Grain, Scales

Mushrooms, Mycology and Psychedelics >> Advanced Mycology

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