Liquid Culture/Mycelium

[Otiliya]

I thought this definitely deserved a thread of its own.
SWIM has read from the giant ocean made out of threads and innumerable copies of same questions on mushies called the shroomery about having endless inoculating liquid and never relying on spore syringes again.

SWIM followed basic instructions by dissolving a small level teaspoon of Karo syrup(honey works too) in about a 100mL of filtered water. Microwaved on high and watched it boil intensely for at least 2 minutes. Quickly covered the jar with alcohol-wiped tin foil and screwed the lid and cap on. The cap already had hole in it for inoculating it with some spores (1mL) when the liquid cools of course.

It has been about 2 days, and it looks like the pic attached. It should grow significantly more in the next week or so.

SWIM has done this before, and had quite some clumps and many little spheres of cotton-like mycelial growth suspended in the jar. The idea was to suck back the liquid into an empty clean syringe and have a solution in it of many suspended mycelial hairs that would be used to inoculate 5-10 more jars and so on. Except SWIM never really had any succcess with this method because one time the mold flakes in the jar just died... He thinks he shook the jar too much/too strong and killed all of it or it was infected without his knowing. The second time he had somewhat of a success with it and innoculated one jar which showed mycelium growing but also something else growing with it. And swim used like half the syringe on that spot.

SWIM has a good feeling about this jar as he was quite sterile, or so he thinks at least and he will also suck up the liquid into the syringe and back out many times to break the mycelium clumps even better. SWIM will inoculate a jar in 3-4 days or so and post success/failure.

Anyone else here has any experience and or tips to share with us who struggle still with this tek?

 

[dread]

One suggestion: learn agar work. LC:s are tricky. For one thing, you can't tell if an LC is contaminated by appearance. Mushroom mycelium and mold mycelium looks exactly same in a liquid culture. Working with agar is easier than people think.

Especially multispore LC:s are tricky. If you insist on using these you should always test your new LC on one jar first before you waste a whole lot of spawn...

That said, LC is a convenient way to make inoculant. Grow a culture on agar first, isolate the best mycelium, then use a wedge of agar to inoculate the LC.

Also, microwave is a bad way to sterilize. There are tons of bacteria in your microwave. A pressure cooker would be best, but if you can't acquire one, steam sterilization will do.

Honey is the best material for LC in my experience. It lowers the chances of bacterial contamination, being a natural antibiotic.

Also, most people put way too much sugar in their LC:s. I've seen LC recipes where you end up with 4% sugar in your LC. This is too much, and will not accomplish anything. 1% sugar content is enough, 2% is the maximum you should use. This means 1-2 grams of honey to 100ml water. Mix the honey and water together, and pressure cook for 20 minutes at 15 psi.

 

[acolon_5]

More is not less with LC's. I usually do 200mL to 1/2teaspoon honey 1/2teaspoon light karo. And LC's are VERY tricky. I've made the mistake of thinking one is good and wasting it on a ton of grain...

Also a microwave MIGHT work, but a better way to do it is to boil your water, pour it straight in your jar with karo/dextrose/whatever already in there, cap and PC for 15-20 minutes. Again less is more, more than 20 minutes and your sugars will caramalize and mycelium won't grow as well.

I never learned to work with Agar, maybe that's my next project..hmmmm.

 

[Phlux-]

swim always uses dextrose/glucose 3% by weight
so 3g dextrose/glucose 97ml water - pressure cooked for 25 mins.

 

[Otiliya]

So foaf of mine did another very similar liquid mycel karo jar. 100mL water with now 1/3 of tea spoons of karo light syrup. Reason for that is he listened the nexus members here. He also boiled the water for a bit in the microwave with the syrup in it already, did everything as sterile as possible with lots of alcohol wipes/covers. Then boiled the closed jar with a little hole in the lid inside a pot, covered tightly so steam and heat would be in plenty inside the pot. 30 mins of boiling. Inoculated with about 1.5 cc or less of spore syringe solution from vendor.

The growth seemed to be faster than the first jar, which btw i think is contaminated, because the mycelium there has a darker tint to it and feels not so fluffy anymore, almost decaying or something. The second new jar *looks* clean and good. Still does. About 2 days ago, foaf of mine decided to suck some of that mycelium goodness into syringe and inoculate a little cake jar. The mycel was broken down plenty by sucking in and out quickly and there was definitely a good deal of mycelial fluffs/flakes/bits floating around everywhere in the jar. That was sucked in the syringe and used about a third of it on one little jar. Inoculated in 2 places. Nothing as of yet.

The cake substrate is actually about 3 weeks old maybe more. It doesn't look contaminated at all, no smell, no funny colours. FOAF of mine actually is good at that, having clean cakes, never had a contaminated cake unless very careless.

Why is there nothing growing? what is going wrong?

 

[Otiliya]

this is the 2nd karo liquid culture jar, and it looks good right? that's how it is supposed to look right? swim inocculated using that jar twice now and nothing! swim used 3mL on the first jar and used the rest 6-7mL on ONE JAR! nothing still. there were plenty mycelium flakes floating in the syringe.

someone please help, what is going wrong?

 

[dread]

it looks good right? that's how it is supposed to look right?

Yeah, but that's the thing with LC:s. You can not tell if they're viable just by looking at them.

From the appearance I can tell you have some mycelium in there. Whether it's mushroom or mold only time will tell. Try it on one jar first.