If you made LC from a colonized jar of grain, could you...

[H2O2]

...use those syringes to inoculate many jars further down the road, at your leisure?

Also, could you then use a similarly made batch of syringes, from the newly colonized jars, to continue the process?

I've read that the strain loses vitality, but, how many generations do you think it'll last?
Three? ...Four? Then what happens?

I ask because, if you got huge flushes, and used very sparingly, you could get a lifetime of mushrooms from one colonized jar. Also, making sterile prints / syringes from prints could be a real PITA.

Do I have any major wrong assumptions here?
Thanks.

 

[Cultosaurus]

A)...use those syringes to inoculate many jars further down the road, at your leisure?

B)Also, could you then use a similarly made batch of syringes, from the newly colonized jars, to continue the process?

I've read that the strain loses vitality, but, how many generations do you think it'll last?
Three? ...Four? Then what happens?

I ask because, if you got huge flushes, and used very sparingly, you could get a lifetime of mushrooms from one colonized jar. Also, making sterile prints / syringes from prints could be a real PITA.

Do I have any major wrong assumptions here?
Thanks.

A) Conventionally, we make LC from the spores. That way in the jar, all we have is the white clouds of fluffy myc and no little bits of organic stuff. However, yes, some ppl have a TEK where they get some myc from the jar, or swish some sterile water around the jar once the grain/BRF is removed to get a solution to use to nocc up the next jars. But yes, take your live myc and put it to LC or agar.

B) yes. That''s exactly how we do it. Once colonized, the LC will continue. 3 generations is safe. Some say 4 or 5, but I'd suggest no more than 3. For furthering your LC, make new LC from a print or tissue from one of your shrooms. This is how you continue on forever, not from LC directly, but from the spores or clones from the shrooms that you get.

I tried to find the name for what happens after successive generations, it's one of those odd mycological terms. HA, I looked again, found it. Senescence.
http://en.wikipedia.org/wiki/Senescence . Biological aging.

Most commonly, you'll lose potency with Senescence, but I have read a few where the shrooms got smaller, but the potency remained the same. I wouldn't count on that happening though. Get a culture and make a new batch.

 

[H2O2]

Thanks.

One more thing please, if you had a fully colonized jar with a set of self healing ports on the lid.
And you injected a lot of sterile water, swished it around while the colonized grain is still in the jar, then sucked it back up...can those syringes last for years in the fridge?

Or will the organic material that you sucked up with the bits of myc somehow contaminate the syringe?


Thank you.
That senescence link was pretty cool, too.
I love science.

 

[Cultosaurus]

Conceivably, yes. You'd just have to insure that you get no contamination. It's organic material that would rot inside your syringe if it had any kind of bacteria on it. For storage for years, it's easiest to refrigerate LC or agar. For long term storage many ppl use what's called a slant. It's a test tube with agar in it, placed on an angle to get maximum surface area, thus, slanted. You'd grow it just like a petri plate, then refrigerate when the agar is covered. You'd have some kind of cork in the top to seal it. Also, with a syringe, you'd have to insure that no contams come in from the needle side.

The best long term storage is a spore print. I have seen some ppl bring a 30 year old print back to life. I don't know the max on LC or agar though. But it does go over a year.

 

[H2O2]

Okay. I can see how traditional LC and agar slants would be more risk-free... once you got them in the fridge.
"Easiest', I'm not so sure about though. Spore prints and agar work seem like another opportunity for contamination.
And, they're a whole new area of study and practice for most people. Another way for an amateur, like yousr truly,
to mess something up. That's why I like the idea of getting the myc strands right from the grain.

You can see that it's healthy and uncontaminated.
And with self healing ports, it seems much more sterile, with less steps and less chance of contamination,
...than doing bench work with spores or agar plates 'out in the open'. What do I know, though? It's all theoretical to me.


All right then, thanks for answering these questions, Cultosaurus!

Feel free to add anything to this thread, anyone at all, I'll be checking back periodically.
All input is much appreciated.

Salut! Ciao! Pass auf dich auf! Thanks!

 

[Cultosaurus]

Prints are, or should not be, too much of an opportunity for contam. Once you have some growing mush, unless you see black mold, green ninja or some other contam in your FC, you should be fine to print a cap. It's easy. Hopefully you'll have a GB, glove box, and can do your work inside there. You just take clean tin foil, wipe with alcohol, break the cap off of the stem and place gills down. If done in a GB, there is no need for this next step. (but cover your arm holes in the GB) Some people do it in the open, same steps as described, then cover with a glass, goblet or jar. Place a coin or something small under the glass to allow moisture out. It will take about 24 hours. Then put the print into an unused ziplock for storage. If kept in a cool dry place, they will last for years.

Quite often wild prints are taken this way and we have to clean them up on agar to get a good culture. That is where you would need to practice some agar work. Many times people just order a print from a spore house and make their own syringe. That would be just about the same as making a syringe from your own print. It's all a matter of confidence level. And everyone gets contamination from time to time. It's not a disaster, you just have to practice and you'll get better at it. Golden rule : "Cleanliness is next to shroomyness". As much as possible, keep your work area clean and sterile ... . many successful growers do much of their work in open air. But a GB or a flow hood really helps.

 

[H2O2]

Yeah, you're pet myc is protected from contamination when it's got active fruit bodies, I'm guessing,
because the mycelial network is strong and the organism is fending off the growth of competitors.

Somehow.
:?

 

[My Pitiful E-Rep]

So with agar work, you take your inoculate (whatever that might be)
introduce it to agar and then separate the healthy myc from any possible contams, correct?


Also, whats the difference between taking a fully colonized grain jar,
shooting, sloshing and sucking up sterile water ...and LC. Wouldn't that be considered a 'LC'?

 

[Cultosaurus]

So with agar work, you take your inoculate (whatever that might be)
introduce it to agar and then separate the healthy myc from any possible contams, correct?

Yes. You can introduce LC, spores or live mushroom to agar to start the growth. It's a good way to keep cultures in the fridge for up to about 2 years in what we call a slant. A slant is a test tube with agar and myc inside and a cork in the top. The myc does not need a continual supply of Oxygen to survive for that long. It's called a slant because you slant it to give greater surface area to the agar inside, you don't leave it standing strait up.

And Yes. Wild prints, the ones we collect in the field are most commonly put to agar to clean up the wild molds and spores that would get on a shroom in the wild. But you may also have to do this with a print that arrived not looking so clean. So, yes about the clean up. When you grow the myc on a plate (actually, I use small masons) you'll have some white ropey myc and if you develop any kind of contam, you slice off small bits of the most perfect white ropey myc that you have and transfer that to another agar petri. You many even have to do this several times until you have a completely clean culture to work with.

Also, remember to work as clean as possible. Working with a flow hood is best, if not, try to use a GB, glove box. Ghetto GBs are cheap and easy if you need to. Very few people try to do this in the open air. Sterilize your knife in a flame.

Also, whats the difference between taking a fully colonized grain jar,
shooting, sloshing and sucking up sterile water ...and LC. Wouldn't that be considered a 'LC'?

LC is a nutrient mixture. If you have used the 'swish' method, all you have is live myc and sterile water. But you can use this swish water to nocc up your LC.

There are many recipes for LC. The simplest is just Karo, as the American's call it. I know it as Corn Syrup. Most people prefer to use the light as it has no colour and makes it easier to see the myc growing inside your jar.

There are all kinds of things to add to your LC. Some are: ME, or malt extract; nutritional yeast; concentrace; and a few other things. Always filter your LC if you add any solids like the yeast. You want your LC to be as clear, even if coloured, as possible. Additives are not necessary, you can get away with just Karo or honey.

Use only 4% sugar by weight, not by volume. ME is a sugar based additive. It's important to remember that things like karo, honey and molasses are not 100% sugar. So, on a recipe for 100 gm of water, you'll be adding just over 5 gm of honey or karo. If measurement is not easy, it's best to have less than 4% than more than 4%.

Other things you can use for LC are honey, molasses, demura sugar, RAW sugar, Turbinado and Muscovado. Any of the brown sugars are less refined and still have some mineral content in them that is a nutrient for the myc. Although Molasses is a great one, it was hard to make out the myc inside the jar of dark liquid.

Never fill your LC jar over 3/4, usually only about 1/2 to 3/4. You want air in the top of the jar. Once the oxygen is exhausted the myc just stops growing. It will survive in the fridge for up to about 2 years maximum. Most only go for 1 year. You can add marbles or small bits of glass to the jar. It will help to break up the clumps of myc when you want to syringe some out. You'll be shaking the jar gently to mix up the myc and break up the clumps.

 

[My Pitiful E-Rep]

Just gonna throw this one out there:

Can't you put a colonized grain jar, properly wrapped, in the refrigerator for years?
Then just do G2G, which I am told, is faster...?

Also, many people have good experiences just sticking to plain grain, with a plain verm casing.
Half gallon jars and turkey trays. In a tub. Simplicity, simplicity, simplicity. I've seen results on other sites.

So, can you see any *simple* additions to the grain , like gypsum, coir, 'pond water' ( or is the whole point of grain to keep the nutrition inside the hull to cut down of contamination, maybe the pond water would 'infuse' the grain with nutrition, putting it inside? As opposed to on the hull where the contams can get at it and start to spread...I dunno), simple improvements in a straight 'oven sterilized' verm casing, or in the process itself...that would benefit this uber streamlined process or increase yields? What would be the next step here..aside from doing straw and cow poo which seems like a total PITA. lol.

For someone moving on to agar work and trying to perfect spore printing and that whole process of connecting the end of the cycle with a new beginning, and wanting to square away the main procedure here.


Thanks.

 

[Cultosaurus]

Can't you put a colonized grain jar, properly wrapped, in the refrigerator for years?
Then just do G2G, which I am told, is faster...?

I suppose that you could try. I've never seen anyone do that for long periods though. Certainly, that's what people do for short term storage, like being interrupted while growing where you won't be home for a while. The problem that may occur though is that you may get rot, contamination or invitro shrooms growing in the fridge. Those surprise invitros are something that people find all the time, when they come across something that they had forgotten about.

Also, many people have good experiences just sticking to plain grain, with a plain verm casing.
Half gallon jars and turkey trays. In a tub. Simplicity, simplicity, simplicity. I've seen results on other sites.

So, can you see any *simple* additions to the grain , like gypsum, coir, 'pond water' ( or is the whole point of grain to keep the nutrition inside the hull to cut down of contamination, maybe the pond water would 'infuse' the grain with nutrition, putting it inside? As opposed to on the hull where the contams can get at it and start to spread...I dunno), simple improvements in a straight 'oven sterilized' verm casing, or in the process itself...that would benefit this uber streamlined process or increase yields? What would be the next step here..aside from doing straw and cow poo which seems like a total PITA. lol.

Coir or Straw is a pretty normal addition to putting your grains in poo. It's added to hold the moisture content and for nutrition. Other very good and simple additives are the gypsum, pond water and coffee. Gypsum adds both calcium carbonate and sulfur that the mushrooms need as nutrients. Pond water has a higher mineral content than tap water and shrooms need all kinds of minerals, such as: copper, selenium, iron, phosphorus, potassium, magnesium, manganese, nitrogen and some others. All of these are very minimal, concidered trace elements, but the more you can provide, the better the shroom will do. Coffee in the form of coffee liquid or the coffee grounds adds nitrogen for the mushroom. You can soak your grains in a 1/4 coffee blend with water to improve nutrition. If you get what's called myc piss, that yellowish liquid on your colonized tray, that is just metabolites that comes from a higher nitrogen content and is nothing to worry about.

The nutrition of the grain comes from the whole grain, from the hull and the inside. Hopefully you won't have any contamination if the PC has done it's job. Soaking in pond water or coffee will add a little bit to the grain's nutrition.

There are 2 kinds of mushrooms, dung lovers and wood lovers. Cubies are dung lovers. Moving on to poo with coir or any other mix is a normal way for getting bulk grows or even larger yield trays. If you're doing any kind of grain, such as corn, WBS or rye, going to poo (or sometimes compost) is just about the only way to proceed. Other than that, you'd have to stay working with cakes.

Yield comes from encouraging pinning, IE, getting more shrooms to start growing; and from having a high enough moisture content to the sub, as shrooms are 90% water. Potency comes from feeding the shroom the nutrients that it needs to convert into psilocybin and psilocin. Remember that genetics is the #1 factor in potency though. You can get very good mushrooms from a plain grain sub (added to a poo, coir, what ever mix). There is nothing wrong with doing it that way. People like to add extra ingredients to make sure that the sub has everything that it needs to provide everything possible for the shroom to produce good potency. Just as you would eat properly but also take vitamins, just to be sure. It's that extra little bit that makes you feel more confident in the outcome.

Also, some leave out the casing and find it unnecessary. Many people still use it. It's a personal preference. It's there to keep the moisture content high.