LC from questionable B+ spore water

[Dipole]

Well I did the most pathetic job of washing spores off a B+ print in a plastic bag that can be imagined. So I have a small amount of barely noticeable spores in 100ml of distilled water in a pint jar w/ injection ports. I probably lost more spores than I gathered. I ain't doing it that way again, I am not coordinated well enough.

The spore count looks anemic to me, but I am more concerned about comtams that must have been introduced by the jackass handling of the spore print and syringe. So my plans for noc'ing up some brf jars with spore water don't sound too good to me right now.

So I am making some Karo/vanilla LC jars instead. I am thinking that I can noc up the jars, contams and all. Two days later I will inject 3ml of H2O2 to kill off the contams.

Does this sound like a viable plan?

I'm thinking I might also place a ring of incense sticks around the jars, get naked, and light the incense with a propane torch while singing the opening music to 2001 Space Odyssey and doing some ballroom dancing moves.


Thanks

[(nilla)]

Ehhhhhh,,,not so much.

ummmm h202 wont kill of the contams
and it is very difficult to spot contams in LC
you spot it right away on agar / grain / brf

your best bet would be to test the syringe on agar / grain jars / brf cakes.. you dont have to use the entire syringe.. just enough to see if it got contaminated.. then if its not, you can finish it up..
who knows? maybe you got a clean syringe.. just try it.. good luck

[Dipole]

Oh. I thought I read that many spores and comtams can be killed or slowed by H2O2. That mushroom mycelium could tolerate H2O2. Hippie even recommended using 1-3cc H2O2 in an LC for storage in a refrigerator.

[~Hippie3]

yeah, well, after more experience i eventually concluded it wasn't much help, massive contams in liquid are impossible to get rid of entirely without killing the mycelia too

[Dipole]

Bummer. Well I am doing 3 pints of LC. Maybe I will get lucky.

[Dipole]

2 out of 3 jars have myco blobs floating on the bottom. The jar that doesn't is potato/karo/vanilla. It was a little warm when it got spore noc'ed. FOAF didn't think it very warm, could this be a reason for a slow start. I has been 5 days, the other 2 had obvious growth in 3 days.

No contams that can be seen.

I didn't bother with H2O2. I will post pics tomorrow if I can.

[Dipole]

Here are pics of a B+ LC started from what foaf thought would be a badly contaminated spore solution.


Karo and Vanilla


Karo and Vanilla


Potato Water, Karo, and Vanilla

The solids from the potato water have sort of settled on the bottom.

Room temperature is in the low 70's F.

Thanks

How much growth should foaf go for before inoculating substrate?

Any problems that I am not seeing?

[vitae1234]

What are your feelings on growing out a test jar? Not to oversimplify, but you're either going to grow out healthy myc, or grow out contams. Unless you want to mess with agar, there's one way to find out.

It would be a shame to noc up a bunch of substrate with contaminated LC, so if the test jar bombs, I'd cut my losses and start fresh.

FOAF can send you a B+ print if your stuff fails.

[Dipole]

Thanks vitae1234 for the spore print offer. FOAF is planning on doing a test run with the LC jars. One of the LC jars is for someone FOAF knows so a good jar is important. FOAF does plan on giving the jars at least 2 weeks to prove themselves contam free.

[dial8]

Looks like all 3 are showing some growth unless the last pic is just potatoe sedimentation. Let them go for several more days. YOu want it pretty thick in there. How many days has it been so far? Usually takes about 2 weeks to have sufficient growht to make syringes from.

Its hard to id contams in a lc that is why you should always test an lc on a jar or two first before you knock up a bunch.

[Dipole]

Thanks Dial8, the jars were 6 days old at the time of the pics.

I have read that contams are hard to see in an LC. Is that still largely true even at 2 weeks?

[dial8]

Yes its true even at 2 weeks.

Trick and other sporulating fungi are the same color as mushroom myc. THey change color upon sporulating and they rarely sprulate enough in an lc to detect the color difference.

Sometimes the myc will look slightly different and bacteria can have a different look but even these things are hard to detect unless the problem is really bad.

Unless you are very confident in your sterile techniques you should always test your lc.

[Dipole]

A test run will be done. When it comes to sterility, confidence is severely lacking. Pics will be posted again this Friday.

Thanks for your help.

[jimbojones]

I'm keeping my fingers and toes crossed!

Good luck Dip!

[~Hippie3]

those all could stand more growth before use.
any air exchange going on, any stirring ?
both improve growth significantly
http://forums.mycotopia.net/showthread.php?t=5177 (Bubbling your LC: airport in action)

[Dipole]

The B+ is still growing. About double from last week

About 100ml of fluff in each jar.

Let it go a another week?

[dial8]

Its looking real good and with that much you could make up some nice syringes but I like mine to be just a bit thicker. Let it go for another few days.

You are swirling it occasionally, right?

[Dipole]

No, I haven't swirled ever. I guess I had better get started. Thanks for letting me know, I didn't have a clue. I thought it was best not to screw with them. Thank you for commenting on the post.

[~Hippie3]

merged 2 into 1 thread

[Dipole]

Wow, swirling brought the myco in lifeless jar to life. I hope it is myco. I plan on noc'ing up some super brf cakes tomorrow. May fortune favor the foolish. I hope the LC is clean.