On This Liquid Culture.....
|Posted by: EvilShroomer Dec 23 02, 08:33 PM GMT|
| ok , on this liquid culture stuff , people say , use honey powder stuff ,
dextrose , sugar water , corn syrup , etc , well , cant we just mix it all togeather , and hopefully get one nice good growth?
|Posted by: Mycota Dec 23 02, 08:55 PM GMT|
Most Honey has some wax in it.
Dextrose is lacking, in certain nutes
Sugar water sucks
Corn syrup, is a liquified dextrose, with some unwanted additives.
My best luck was with DEXTROSE (corn sugar powder) & a LIGHT DRY MALT (powder). Both available cheap @ almost any home brew store. (Just don't PC or nuke until it carmalizes into a dark brown)
There is no real STANDARD solution.
You want the solution CLEAR, so you can see what's going on and/or if contams appear.
Most failures with liquid culture, are either a lack of OXEGEN (causes it to STALL), a bum mix, or simple contams.
1/2 teaspoon of dextrose & 1/2 teaspoon of light dry malt, well mixed in a quart jar, PC'ed or nuked, then split into 2 quart jars (leaves room for OXEGEN), PC'ed or nuked again, allowed to cool to room temp, then innoc'ed with spores, almost never fails for me.
Once you have some thick myc growth, stick the sucker in the fridge, to stall growth. It will store several weeks. Then, take it out, let it get to room temp, when you want to use it & innoc whatever.
|Posted by: dankcrop Dec 23 02, 10:09 PM GMT|
|i just made some karo syrup sunday morning. Should i loosen the lid and take the tape off the holes for more oxygen? I dont want this jar to stall.|
|Posted by: Mycota Dec 23 02, 11:25 PM GMT|
If 1/3rd, to 1/2 the jar space.....is air? Don't bother, loosening the lid or removing the tape. At least until you see some myc, growing in clumps.
If the jar is plum full of liquid, then, cover the lid with alc swabbed coffee filters - held down w/rubber bands & unscrew the lid, a bit. Just to allow some air - in.
|Posted by: dankcrop Dec 24 02, 12:12 AM GMT|
|I used 100ml of water in a half pint. So ide say 1/2 half is aair. Thanks for clearing that up Mycota.|
|Posted by: Nanook Dec 24 02, 12:16 AM GMT|
|Loosening just invites contams|
|Posted by: phillinwierd Dec 24 02, 02:33 AM GMT|
| Hey Mycota, how long do you cook (pc press and time) that mixture for, and what is a "burn" mix? Also, is there a ratio of the colonized liquid to grain spawn recommended for a quick run? Curious to try this one.
|Posted by: OZZ Dec 24 02, 07:34 AM GMT|
| I just always use the standard Karo tek and havent had problems with it contaming or stalling once. 1 teaspoon of Karo light Corn syrup to 100 cc's distilled water in a 1/2 pint wide mouth. PC'd for 20 minutes at 15 PSI allowed to cool and inoculated, incubated and in about 8 days its ready to shoot. I have never worried about loosining the lids and have never had them stall on me or contam. After shooting jars just throw the rest in the fridge like Mycota suggested and your good to go.
Easy peasy japanesey
|Posted by: dankcrop Dec 24 02, 09:39 AM GMT|
|I prep'd mine early sunday morning. Still no signs of growth. I suppose it takes from 3-5days for germination?|
|Posted by: Fungusmaximus Dec 24 02, 09:46 AM GMT|
|Temp affects germination, make sure your sitting @86 F. And put that puppy away for a few days in the dark, forget it. Next time you check there will be no doubt if its working or contam'd.|
|Posted by: phillinwierd Dec 24 02, 10:36 AM GMT|
|OK then Ozz, it's Karo then. Will contams appear as they would on an agar plate or a cake (nasty green fuzzy, stinky shit?) Pictures would be cool if anyone has any to share. And I'm still unclear as to what a burn mix is. Any help? Also I was reading somewhere about the use of bee pollen with this tek but, unfortunately, the thread got contaminated with other issues which made the reading kinda tough. Any takers? Merry Christmas All. Nuthin' but Peace and Love.|
|Posted by: Mycota Dec 24 02, 12:02 PM GMT|
I agree Nan that loosening invites contam's. I use special filters, installed on the lids of liquid culture jars. But, most don't have such things.
That is why I mentioned, you can swab a jar, then wrap alc swabbed double / triple coffee filters - over the jar lid & secure it to the jar with a thick rubber band.
Then loosen the lid.
Those coffee filters act as external filters, to help keep contams - out - if you loosen a lid, a turn.
|Posted by: Mycota Dec 24 02, 12:51 PM GMT|
The term I used was "BUM" mix. Not "BURN" mix. Meaning, screwed up, no good, dark & cloudy, has particulates, to much of one thing, or another. Just a BUM mix, as in NO GOOD.
Here is a picture (close up) of a light dry malt & dextrose mix. The liquid is AMBER colored, like honey. There are green plate glass shards in the jars bottom (to help break / cut myc when the jar is swirled). The green glass gives the jar a funny tint.
|Posted by: Mycota Dec 24 02, 01:14 PM GMT|
| Quart jar of fully colonized seed/grain shot up with that liquid culture @ 14 days. Dark spots against inside of jar are not contams, they are seed/grain against the glass.
|Posted by: OZZ Dec 24 02, 01:36 PM GMT|
| Good pics Mycota, sorry guys I dont have any pics of karo contams because Ive never had one contam with that method of prep. LIke i say 1 teaspoon light Karo corn syrup to 100 cc's water, pc for 20 minutes, inoculate, incubate for 7 - 10 days and your good to go.
This is a pic I took from the archives of Karo at 1 week old. Its about ready I would let it go another 3 days or so from this point, shake the living hell out of it and your ready to shoot jars.
|Posted by: OZZ Dec 24 02, 01:39 PM GMT|
| The point to emphasize here is that you really want to shake the hell out of it before you use it on jars to be colonized, you dont want the big clumps like in the picture when your using it (however while it is being incubated its good to let it grow together like this) by shaking the hell out of it you break up the myclieium into thousands of minute pieces so you dont have any dead spots in your inoculum.
HOpe this helps
|Posted by: repobob Dec 24 02, 02:27 PM GMT|
| Thanks to Nan I used Visions NY tek. Just add 1/8 tsp Neutrional Yeast to your HW or Kayro. Don't shake. I shot this jar up on 12/4 and they have been in the fridge for a week.
|Posted by: Mycota Dec 24 02, 03:29 PM GMT|
| That looks good. Mycota is off to get some nutritional yeast & give that one -- a try.
Who says you can't teach an old dog, a new trick.
Oh yah. One other thing, one time just screwing around.
Quickest liquid culture, I ever did was to spoon in a few spoon fulls of fully colonized seed/grain, into a quart jar half full of sterile water (under quasi/clean room conditions of course), add cap, shake the sh*t outa that puppy, let it set overnight, shake again & used fluid from it as an innoc. It worked FINE.
|Posted by: phillinwierd Dec 24 02, 03:31 PM GMT|
| Oops, guess 25 years of welding DOES take its toll on ones eyes.
Bum makes a lot more sense than burn. Guess I better start wearing my reading glasses. Anyhow, I'd like to thank everyone involved in this discussion on liquid cultures.... very informative, great pics. I'll be giving this a shot as soon as the holidays die down a bit. Lost my partner earlier this Fall. He (biochem major extraordinaire) took care of the petri cultures (over 30 of them), autoclaving grain in bags, and grain innocs., while I (the country boy dead head) did the composting, subtrate runs, casing, fruiting, harvesting and drying. Worked great until well..........we fell out.
Wish he wouldn't have taken ALL the cultures. On the bright side, I still have all the wherewithall and the inclination to get going again.
P.S. The glass shards sound like a great idea. I gather the mycellium tends to get a little clumpy eh?
|Posted by: OZZ Dec 24 02, 03:36 PM GMT|
| Ya Ive been anxious to give that nutritional yeast a try !! I read that at Mycotopia awhile bag, great, interesting thread for sure !!
|Posted by: repobob Dec 25 02, 12:39 PM GMT|
| Have to try that Mycota. Never though of using grain to innoc liquid, but hey Mycilium is Mycilium and it just wants to grow. Neat idea. That gives me another backup on the MexA
|Posted by: Fungusmaximus Dec 26 02, 01:59 AM GMT|
| Yeast is good, great even.
Only thing is it makes the liquid cloudy, making it harder to spot contams.
Other than that, its a winner.
|Posted by: phillinwierd Dec 26 02, 08:33 PM GMT|
| hey hey. Followed your directions for the Karo mix. Made 4 1/2 pint jars. 2 I innoculated with a Mex B print my son and I brought back from Amsterdam last month (great f*ckin trip by the way), and the other 2 I'm experimenting with. Mycota got me thinking when he innoculated a liquid with colonized grain, so I shaved some mycellium off the side of a cake from PR spores. Just wanted to see if it would go. Kinda neat because the shavings float. Interesting to see if the mycellium will grow there too, assuming it grows at all. Plan on shooting some grain with favorable results. Hell, maybe I'll take a cake and throw in the Everbach and make a solution. One of my girl friends got a digital camera for X-mas so I'll take some pics and send them along if anyone is interested.
|Posted by: Nanook Dec 26 02, 10:21 PM GMT|
|Posted by: Molester Dec 28 02, 01:44 AM GMT|
|So, do I need to blend up my dextrose solutions before sucking it into syringes or will a good shake do the trick? Last time, I used the blender and got lots of rust (my blender is about 14 years old)|
|Posted by: Nanook Dec 28 02, 02:34 AM GMT|
|Don't blend it. Get in with the tip of the sterile syringe needle and whip at it a bit.|
|Posted by: phillinwierd Dec 28 02, 08:20 PM GMT|
| Well, the Karo jars that I put the mycellium from the cake into on the 26th have white wispy stuff all through them. The two jars I innoculated with Mex B spores on the same day aren't showing anything. But it's only been two days. Stands to reason that an established mycellium would go faster than a spore that needs to germinate, right? Keeping them with the pots and pans in a cupboard above a heating vent where it stays about 85 degrees and dark.
Anyhow, my friend is supposed to bring over her digital camera tonite. I'll try to snap some pics (Shhhhhh, she has no clue). Good God, now I'm going to have to figure out how to post them. Don't even know how to post an avatar. Hell, I mispelled my board name (four in the morning and nine eyed when I registered) and I don't know how to change that either. Soooooo embarassing. It's a wonder I even found my way here!
|Posted by: Nanook Dec 29 02, 12:47 AM GMT|
| Yeah spores take a few days longer, 3-5 day head start.
|Posted by: ion Dec 29 02, 01:06 AM GMT|
| Well, I'm just confusd as to why everyone keeps talking about "nutritional yeast". Where I'm from, the only thing around is "Brewer's yeast". I'm assuming it's the same stuff, but I could be wrong...
But I guess I'll find out soon!
Pics to come.
|Posted by: repobob Dec 29 02, 07:36 AM GMT|
| Ion, "nutritional yeast". and Brewers yeast are different, I think.
NY can be had at health food stores. Its like CornFlakes only way thinner.
|Posted by: phillinwierd Dec 29 02, 11:52 AM GMT|
| Day three: Jars innoculated with cake mycellium are really taking off! Jars innoculated with Mex B spores have several small, white puffs about the size of peas. I'm assuming it's mycellium. Wish my friend wouldn't have forgot her frickin' camera. Anxious to post pics and get some feedback from y'all. Anyhow don't see anything foul in there yet. I'm gonna start putting my flowhood back together now. Wasn't real comfortable using the oven tek to innoculate despite the absence of contams. More comfortable sitting at the table in front of hood instead of on my knees at the oven.
|Posted by: ion Dec 30 02, 03:02 AM GMT|
| That's just it, bob... it seems that no health food stores carry anything called "nutritional yeast", only the brewer's yeast. Thing is, brewer's is a flake form yeast that is sold as a nutritional supplement. It is soluble in water (to a point) and has lots of B-vitamins, apparently. Like I said, maybe it's just a regional thing.
|Posted by: repobob Dec 30 02, 05:04 AM GMT|
| Oh I see what you mean Ion. I'm not sure if they are the same. I read something where there is one kind of yeast that is "live" ... that not what you want. If its a neutrional supplement I would assume if not the same thing its awful close.
|Posted by: phillinwierd Dec 30 02, 03:32 PM GMT|
| Just went to the health food store to get some nutritional yeast for more liquid culture. They had nutritional yeast and Brewers yeast in powder and flake forms. Asked the woman what the difference was between the two. She told me that it is basically the source of the yeast.
Nutritional yeast is a primary grown yeast grown on enriched purified cane and beet molasses from some critter called Saccharomyces cerevisiae.
Brewers yeast is a bi-product from the fermenting process used for beer. (Mmmmm beer) It may contain unwanted critters for use in mycology (my assumption, not her words), most notably Candida albicans yeast.
So, my laymans take on it is that there is a difference. Nutritional is prefered over Brewers.
I'm sure one of these edumacated geniuses on here could easily give you the scientific fine points on the matter. They freak me out with the extent of their knowledge (try reading the thread on peroxidizing mycellium).
|Posted by: 420M Dec 30 02, 06:16 PM GMT|
| I've seen nutritional yeast at my local Vons.
Good Luck - 420M
|Posted by: repobob Dec 30 02, 06:35 PM GMT|
| Looks like there is a difference between the two.
Ion, if you can't find any PM me and I'll get you a container of NY. You only use an eigth of a teaspoon, so 1 container will last forever. That makes me wonder how long the stuff stays fresh. It came in a cardboard container like Oat Meal. I think there was like a lb in there.
|Posted by: ion Dec 31 02, 12:54 AM GMT|
| Hmm... perhaps a trade is in order.
|Posted by: phillinwierd Dec 31 02, 02:09 AM GMT|
|Mine came in a plastic jar. 10 ozs. for $10.00. You're welcome to some if you need it.|
|Posted by: ion Dec 31 02, 02:30 PM GMT|
| Much appreciated, folks. I've been able to find the stuff on the net, though. Thanks, anyway.
I've done a little searching and come up with relative figures on nutritional information for the two yeast forms.
Apparently, the leading NY "Red Star" is a non-bitter yeast that has been enriched with B-vitamins (hence people's assumptions that it is more nutritive). The only real differences between the two yeast forms are as follows:
NY is less dense than brewer's. 2 tbls weighs about 15 g, whereas 2tbls of brewer's weighs 30 g.
The nutritional content is almost exactly the same (accounting for densities), except that brewer's contains more magnesium... this might suck because myc doesn't generally like magnesium from past research... though small amounts are necessary for healthy growth.
NY is less bitter than brewer's... this really only makes a difference to those that are eating it.
NY is made in a lab, and brewer's is made in a brewery (usually). They are different yeasts, but have very similar nute values. No candida in brewer's, either
|Posted by: Workman Nov 11 02, 01:29 PM GMT|
| The impurities are likely adding to your success with pure sugar solutions. Mycelium needs trace elements and a nitrogen source to thrive, which are sorely lacking in carbon rich sugar solutions. I am suprised that there are any successes using pure dextrose or even relatively pure corn syrup. Also the sugar to water ratios are a bit high and can cause osmotic stress to the mycelium cells which will slow growth.
A few years ago I tested a series of dilutions of potato dextrose broth to see what the dilution was for best growth. I started with the below stock solution.
Stock solution (100%)
250ml potato water from boiled potatoes
1 tsp dextrose powder from the brew store
I then did rough serial dilutions of 50%, 25%, 12% and 6% as shown in the image. The blue things are magnetic stir bars. The jars are illuminated from the bottom to show detail.
Suprisingly the 25% dilution was the thickest and fastest growing, but even the minimal dilution of 6% had good growth. More than enough to be used for innoculation purposes. It might even be better since the thinner mycelium is easier to draw up into a syringe without clogging and doesn't form the thick mat on the surface. The 25% eventually formed mushrooms on the floating mycelial mat, but I didn't take a picture
Currently I use a teaspoon of malt per 500 ml of water in a quart jar. Its a bit thick when colonized (like jello) but its an easy measure and it works fine for my purposes. Malt, like potato water is unrefined and contains all necessary elements for healthy fungal growth. It is no accident that you don't see recipes for honey water, dextrose or karo agars.
That's the Nook Shroom Glossary