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Topic Author Last Poster Posts Pages Last Post
Liquid Culture - 1
Smelly honey tek onenessMarx2k6 1 10-27-01  09:30 pm
Honey jar contam or mycellium?Mr. Tambourine ManMr. B19 3 12-13-01  10:43 pm

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C K (Phrozendata)
Posted on Sunday, November 04, 2001 - 09:25 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

This question came up and got me wondering, expecially if i'll be making some soon. I'm hoping to become self sufficient with my next crop. Spores will be taken on tin foil and wax paper and used to make syringe. Other than doing a "two jar test" is there any methods you guys use for testing the sterility of syringes?
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Nan (Nanook)
Posted on Monday, November 05, 2001 - 12:06 am:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

I shoot a jar of Liquid Culture Tek, Dextrose specifically, with 1.5 ccs of spores and let it incubate for 5-10 days. Then you will know for sure if the spores are good, and if the liquid culture pans out you can shoot the mycleial solution
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C K (Phrozendata)
Posted on Monday, November 05, 2001 - 05:47 am:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

How do you know for sure? Smell?
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Nan (Nanook)
Posted on Monday, November 05, 2001 - 09:15 am:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

Molds form dark spots. Bacteria discolors and/or clouds. Yeast bubbles. If the culture is clean it looks like fine white puffs of tissue clumping on the bottom of the jar. It's a cheap easy way to test syringes, and if they test good you can go ahead and shoot the mycelial culture.

The jars should be sealed. No smell. No leakage when shaken or swirled. No contams entering. The only way to smell them would be to open and stick your nose in... If you do that the jar will contam and you put yourself at personal risk with contam exposure.
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Nozza (Nozz)
Posted on Monday, November 05, 2001 - 05:56 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

Agar makes it very easy
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Nan (Nanook)
Posted on Monday, November 05, 2001 - 11:33 pm:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

Not for shooting jars it's not. And it's worlds slower. Dextrose is _fast_.
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An guy (Boomer)
Posted on Monday, November 05, 2001 - 12:56 am:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

does that mean it's not mycelium?

One of my jars, the white stuff looks like the stuff in the other jars- very ephmeral, almost like I don't know, just very diffuse, see-through- looks like that in the other jars too, but in this one, it's manufacturing air bubbles, trapping them, and floating to the top.

I keep waiting to see if it will grow out of the water, I read cobweb grows on top of the water, but it is so far staying under the water. Just the air bubbles trapped in it are making it float.

Anyone know what this is? Had this happen to them?

Thanks

boomer
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Nan (Nanook)
Posted on Monday, November 05, 2001 - 01:32 am:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

Yeast infection. Yeast is fermenting the sugar and making alcohol and C02. Fizzling or bubbling liquid culture is a dead giveway for this contam.

The wispy, emphermal stuff that stays on the bottom until you swirl it what a good clean culture should look like.

This is a good place to drop another tip since people are using this tek. Make sure the jars and everything are crystal clean... Not that a bit of filth hurts the tek (because it is sterilized) but the flecks of dirt in the clear liquid culture _look_ like contams... and you never seem to notice them until after the mycelium has started to take off... You will be swirling in front of a good light and all of the sudden there is a flake of verm or a small grain of rice flour that somehow got in there and then it seems to make all the difference, it's very noticeable.

I was embarrased to find a hair and another fleck of filth floating in one of my Karo jars the other day. Of course they are sterile, I forgot to wipe the jars out before filling...
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Nan (Nanook)
Posted on Monday, November 12, 2001 - 04:48 am:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

You can get the full range of contams in 4% sugar solutions. The two most common are yeast and green mold. Cobweb will grow in them, so will bacteria (which typically come along with the yeast).

Incubation time for contams is 5 days. I think that because the 4% sugar solution is so nutrient specific it may give spores an edge... With the exception of yeasts.

Yeast in 4% sugar causes the solution to bubble.

Mold appears as growing dark specks in the solution, they sometimes float to the top.

Bacteria causes the solution to grow cloudy, sometimes clumps and dots that stick to the glass appear. They can (like mold) be any color but they are typically opaque milky, pink, brown, or black.

Pure mycelium culture looks like clumpy threads of white tissue paper that collect on the bottom of the jar. Jars should be swirled frquently to keep the mycelia stirred and broken up.

Try to keep the jars very clean when making them up. Dust, particles, hairs, ect. are easily mistaken for live contams (if sterilized properly they should not be a problem).
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Brad (Raze)
Posted on Tuesday, October 30, 2001 - 02:56 am:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

How do you go about recognising contams with honey tek?
Mine is just starting to colonise. Little floaty strings all over. How would I tell apart shroom mycilium from say mold?
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Nan (Nanook)
Posted on Tuesday, October 30, 2001 - 03:45 am:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

Mycelia is white, pure white and looks kinda like stringy clouds floating in a liquid culture. Contams are off color, none of them look like stringy white clouds floating in liquid culture.
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Brad (Raze)
Posted on Tuesday, October 30, 2001 - 03:49 am:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

Its hard to tell whats white when the water is yellow...
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Nan (Nanook)
Posted on Tuesday, October 30, 2001 - 04:05 am:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

I am getting to hate Honey Tek more and more with each passing day... Sigh... And it is not you, honestly, it is the honey... For precisely this reason.

Brad, honestly, if you followed the Tek (and I know you are a smart fellow) the odds are greatly in your favor that the stringy floaties you are seeing is a pure mycelial culture. If you have doubts, give it a few more days in the Incubator and if it is contamed, it will look it. You will know.

On the other hand if it is indeed a pure mycelial culture, as I think it likely is, it will fill out and cloud with a large puff that looks like cotton soaked in the jar.

Contams are not likely to hide as the growth progresses: they go black, green, brown, gray, the culture fizzes and bubbles, scums over, crap floats up... It will look nasty and you will know.
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Nan (Nanook)
Posted on Tuesday, October 30, 2001 - 09:18 am:Edit Post Quote Text Delete Post Print Post Move Post (Moderator/Admin Only)

>>since we're on the subject, cloudyness in the jars are ok right? i have a few jars and they're growing but some of them are cloudy and others are clear (used dextrose). they all have puffy "cottonballs" forming in all of them. maybe i'm paranoid.<<

Humm... They should be clear with defined "puffs" of mycelia growing. General cloudiness or haze speaks to me of possible bacteria contam.

>> and black specks are nothing to worry about either right? i mean, those are the spores.<<

Wow... If the black specks went into the jar when inoculating they were spores... But tiny black specks in liquid culture is also a danger sign of cobweb contam... Cobweb fruitbodies in liquid culture appear as tiny black specks. Nothing you want to shoot into jars unless you are _sure_.

The dextrose tek I posted is "pain in the ass" detailed... PC sterilization, glove box for inoculating, solid lids... I had tiny black specks once using Polyfill lids... No problem, spores I thought... I shot 30 jars with cobweb, then birthed, ground the cakes and spawned straw. It was a real mess to clean up, I had cobweb everywhere and it took me days to trace it back to the contamed dextrose jar... When I post specific and detailed Tek info... I do it not to make life difficult for others, I do it to make it easier.

Really.

>>finally, for some reason, i stored some syringes in the fridge and when i looked at them today, the myc was starting to grow in the syringe!<<

Spores in sterile water do not germinate. Period. There is no food unless you loaded the syringe with spores in dextrose or honey.

>>i should use them asap huh?

If you are sure the syringe contained nothing but sterile spores and liquid culture media... If the spores were in pure water I would not shoot.

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