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Offlinekjb1891
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Growth w/ Liquid
    #2155677 - 12/03/03 05:03 PM (13 years, 4 months ago)

When using liquid mycelium syringes, etc. I read that jars usually colonize in 1-2 weeks. How soon do you usually start seeing growth begin though? Does it look the same as if spores were used? Also, when you make a liquid mycelium syringe, how does it look? Is it supposed to be mostly clear?


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InvisibleZwieback0
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Re: Growth w/ Liquid [Re: kjb1891]
    #2155850 - 12/03/03 05:47 PM (13 years, 4 months ago)

It can take up to a week.

Yes it looks the same.

If you used honey tek, yellowish tint and stringy myc should develop.


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Offlinekjb1891
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Re: Growth w/ Liquid [Re: Zwieback0]
    #2156312 - 12/03/03 08:46 PM (13 years, 4 months ago)

I take it that it grows very rapidly once it starts then? So, it would start showing growth by at least a week, then finish colonization winthin a week?(Or so)


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Re: Growth w/ Liquid [Re: kjb1891]
    #2156340 - 12/03/03 08:59 PM (13 years, 4 months ago)

Quote:

Is it supposed to be mostly clear?




That would depend on how potent it is. If you take all the mycelium off a petrie dish and stuff it into a 10cc syringe like I do there will be a lot of little chunks of mycelium in it. It doesn't need to be this potent -- as long as there are cells in it. Big chunks get stuck in the syringes, though, unless you use like a 16-gauge needle.

Speed of colonization depends mostly on the potency of the syringe and how much you put into your substrate and how airy the substrate is. If you inject your inoculum into the middle of the cake, though, colonization can start mainly from the inside, so it can take longer to see anything, but then you'd know when the cake is most likely fully colonized....

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Offlinekjb1891
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Re: Growth w/ Liquid [Re: micro]
    #2156744 - 12/03/03 11:10 PM (13 years, 4 months ago)

It's being used on grains, not BRF or anything else.


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Offlinekjb1891
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Re: Growth w/ Liquid [Re: kjb1891]
    #2159221 - 12/04/03 09:16 PM (13 years, 4 months ago)

I was wondering because the Poor Man's Mycelium Syringe Tek says that colonization should be finished in 1-2 weeks, but after 4 days my friend doesn't see any growth yet.


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Re: Growth w/ Liquid [Re: kjb1891]
    #2159249 - 12/04/03 09:26 PM (13 years, 4 months ago)

If you know there was live mycelium put into the jars and the water content is right and you didn't overheat the jars or add chemicals you will see growth eventually.

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Offlinekjb1891
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Re: Growth w/ Liquid [Re: micro]
    #2159657 - 12/04/03 11:27 PM (13 years, 4 months ago)

So, what's the point in making mycelium syringes when it doesn't seem to be growing any faster than spore syringes? Basically just another way to grow shrooms other than with spore syringes?


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OfflineBlue Helix
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Re: Growth w/ Liquid [Re: kjb1891]
    #2159697 - 12/04/03 11:43 PM (13 years, 4 months ago)

I've been growing jars for 10 years, and with all due respect, these guys don't know what they are talking about with respect to a good set of jars IF they are saying it will take 2 or 3 weeks every time. Liquid mycelium is many, many times faster than spores and is about 1/10th as likely to contaminate since you can use hydrogen peroxide in your mycelium solution.

If you don't keep your jars at an even and the correct temperatures (84-86F), they won't grow as fast. If the grain was too dry, they stall. If you didn't use hydrogen peroxide in your mycelium syringes, it'll grow more slowly. If you used a tiny bit of innoculant rather than a good 4 to 10ml, it takes longer. If the grains were packed tight or weren't expanded enough with water (for birdseed), they'll grow more slowly. There are a LOT of reasons and ways to slow things down.

For me a perfect set of jars using about 5-10cc per jar ALL FINISH--yes, I said FINISH--in 6 to 12 days. If you do it right, you can see mycellium running in the jars in 24 to 48 hours. That's if you do everything right. I mean keeping the jars exactly at 84-86F with a PC fan or whatever keeping the air moving and a dessicant in there to keep things outside the jars dry.

If you do things halfway right, you should see growth by day four at the latest. The jars should finish up by the third week at the VERY latest.

If you don't see growth by day five, forget it. Toss the jars and start over. If you are that far off the mark, your jars will be so far in the balance of bacteria at that point, that even if you DO somehow fix the problem, they'll finish up so slowly they are bound to be weakly colonized. The only way you could expect to fruit such weak jars would be without a casing.


Listen, talk is cheap. I'll post some pictures in a few minutes. Let me get them together. I got a set on 84 hours (3.5 days) where most of the jars are about 75% done.


Edited by Blue Helix (12/05/03 02:33 AM)


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Offlinekjb1891
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Re: Growth w/ Liquid [Re: Blue Helix]
    #2159785 - 12/05/03 12:22 AM (13 years, 4 months ago)

Thanks for the info, but I'm fairly experienced with growing. I've just never used any kind of liquid culture before. I've always used spores. So, I'm really only a nubee to liquid cultures. I did the same size jars of popcorn that I usually do that take 3 weeks with spores. I guess I was expecting too much too soon from the mycelium.


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Offlinekjb1891
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Re: Growth w/ Liquid [Re: kjb1891]
    #2159803 - 12/05/03 12:26 AM (13 years, 4 months ago)

By the way I used 2cc of mycelium solution per pint jar. Is that too little? What kind of spawn do you use Helix?

6 to 12 days is pretty impressive though.


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OfflineBlue Helix
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Re: Growth w/ Liquid [Re: kjb1891]
    #2159826 - 12/05/03 12:36 AM (13 years, 4 months ago)

Here are some photos of a group I got going that are on their 84th hour.

This chamber is a double rubbermaid container (one stacked in another) sitting on reflective bubble wrap stuff you can get at a hardware store. The top of the rubbermain, when closed, is covered with a standard comforter blanket. Keeping the chamber well insulated in very important.

The bottom container has two bricks in it, about 6 inches of water, and an Ebo Jager aquarium heater (didn't show that because I didn't want to move the top chamber around for the picture with the jars stacked right now). The top chamber has the jars elevated about 5 inches on some eggcrate material. There is a PC fan running, blowing down but elevated about 2 inches off the bottom, to keep the air moving against the warmed bottom and spread the heat throughout the chamber.

There is a desiciant to keep the chamber very dry since you don't want any chance for mold or standing water in there (and it DOES take water out of the top vermiculite layer too, keeping it from sprouting cobweb or other molds). NONE of the jars have taped holes. I never tape holes. You have to tape holes only if you let your jars sit around in moist environments and keep mold all around them (I don't do either of those things).

The KEY to this chamber is that the temperature inside is kept even and ideal all around the jars. The bottom jars are not any hotter than the top ones like in many aquarium heater chambers I've seen. The whole chamber can be tuned to a very specific temperature range that seldom varies more than 1 degree.










Edited by Blue Helix (12/05/03 01:19 AM)


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OfflineBlue Helix
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Re: Growth w/ Liquid [Re: Blue Helix]
    #2159885 - 12/05/03 12:58 AM (13 years, 4 months ago)

kjb1891, the amount of inoculate determines how many places it touches as does the dryness of the mix. A too dry mix of birdseed and vermiculite will tend to absorb the inoculate before it touches around many places, even if you do use a lot (say 5cc). Just remember each place the liquid touches will explode in growth. And remember that a single agar plate can EASILY be enough for a HECK of a lot of water. I use a single agar plate with about 1 pint of water. I mix in one half pint. When I'm done, I mix in another using the same agar pieces. Remember just a couple cells of mycelium is all you need. You don't need a 7-11 slurpy in there.

I make my mycelium syringes using agar media and some dry spores. I pour the media in 1/2 pint Kerr jars while it's hot and after it's been sterilized with the jars. I scrape spores off into the agar and close the jars.

Once the mycelium takes off, I make the mycelium syringes by breaking up the agar just like everyone else, but something I do that I seldom hear others doing is I LIBERALLY use hydrogen peroxide in the mix. In the batch you see above, I poured 20ccs of pure hydrogen peroxide over the mycelium chunks. I rolled that sizzling, cleaning hydrogen peroxide around in the jar a second and then added the rest of the water to the jar. The hydrogen peroxide makes it so that if spores get in the solution while you are messing around, they die. It also reduces the bacteria in the solution drastically, giving the upper hand to the mycelium. There are those that say hydrogen peroxide will slow down the mycelium in the first couple days. That is bullshit. It SPEEDS up the jars by giving the mycelium a clear field to run the ball so to speak.



Edited by Blue Helix (12/05/03 01:02 AM)


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Offlinekjb1891
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Re: Growth w/ Liquid [Re: Blue Helix]
    #2161934 - 12/05/03 04:43 PM (13 years, 4 months ago)

I made my mycelium syringe from a colonized cake, not from agar. So, maybe that makes a difference, too?


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OfflineBlue Helix
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Re: Growth w/ Liquid [Re: kjb1891]
    #2162227 - 12/05/03 06:03 PM (13 years, 4 months ago)

Colonized cakes from spore syringes have tons of bacteria in them and many more bacterial endospores. Even the spore syringes themselves have a tiny bit of bacteria, which is why they don't do well if you try to use them to innoculate agar (you see, agar, unlike grain, does not favor mycelium over bacteria). Unless you kill off that bacterial load when you make your mycelium syringe from it, it's a bad start with the bacteria in the lead. If you are going to use a colonzied cake to make mycelium water, I think it's even more critical that you use straight-out-of-the-bottle hydrogen peroxide solution to kill the bacteria for about a minute and then dilute that solution down to a 1:10 hydrogen peroxide from the bottle to sterile water. Otherwise you'll just have too much initial bacteria in there for the mycelium to get a hold of the new cakes.

The only reason I like agar is because you can tell when it is bacterial free. With colonized cakes, there is a lot of bacteria in there, you don't know it, and that's bad news if you are trying to use that cake for another expansion into grain (i.e. using it to make mycelium syringes).

The biggest proof of bacteria gone wild isn't little girl bacteria half naked at the beach. It's a terrible smell. Do you smell that?


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OfflineRockGut
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Re: Growth w/ Liquid [Re: Blue Helix]
    #2276771 - 01/27/04 07:09 AM (13 years, 3 months ago)

Awsome explanations. Thanks Blue Helix.


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OfflineShrumester
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Re: Growth w/ Liquid [Re: Blue Helix]
    #2277330 - 01/27/04 12:44 PM (13 years, 2 months ago)

Wow! That is a perspective I have never heard or thought of before-H2O2 SPEEDS growth. I gave up on H2O2 a while ago after seeing how slow colonization took. I was using it in the substrate, though, not the mycelium syringes. Have you done some controlled experiments to verify this. I would love some numbers (i.e. number of days w/ v.s. w/o). Thanks Blue.


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Anonymous

Re: Growth w/ Liquid [Re: kjb1891]
    #2277377 - 01/27/04 12:57 PM (13 years, 2 months ago)

can you give your recipies guys and sterilization procedures


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Re: Growth w/ Liquid [Re: RockGut]
    #2279067 - 01/27/04 10:59 PM (13 years, 2 months ago)

I currently use about 10-20 ml of hydrogen peroxide straight from the bottle per half pint of water. I add the hydrogen peroxide to the mycelium-covered agar straight, shake it up real well, and then add the water. The mycelium agar, in half pint jars, is sometimes stored for months in the refrigerator before I use it, so it's not always super fresh. This technique has never failed me, and I never have bacterial problems.

Before I used the hydrogen peroxide, I would sometimes have bacterial problems. Also I have noticed that the hydrogen peroxide wash prevents any contaimination during the injections from mold spores in the room. I don't even worry about the room being super clean with hydrogen peroxide washing. I also don't worry about the time it takes to break up the agar since it is washed with hydrogen peroxide. It really just makes the whole process less stressful since I don't happen to own a clean lab to work in.

My colonization times are typically about 7 days for the full set of jars, which are stored at about 84F innoculated with 5-10 ml of mycelium-water-hydrogen-peroxide solution.


Edited by Blue Helix (01/27/04 11:07 PM)


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