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Offlinezeegos
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Peroxide Substrate
    #5650133 - 05/19/06 12:59 PM (17 years, 8 months ago)

I was just reading up on the Peroxide Agar Tek and how hydrogen peroxide kills all bacterial spores apart from birthed spores and living mycelium.
My question is, can you make the standard Pf Tek substrate but in place of water, use a percentage of peroxide solution (same vol of cource) and just inoculate with a small piece of colonised substrate?
This would be good as the peroxide would do all the sterilization work for you and let you relax about having to have a completely sterile enviroment.
If this could be done, what % H2O2 would you suggest?


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Invisiblecreamcorn
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Re: Peroxide Substrate [Re: zeegos]
    #5650159 - 05/19/06 01:07 PM (17 years, 8 months ago)

tried it a while ago for shits and giggles with a standard 10:1 soultion. failed. got mean green in a major way.... it took a long time for the green to show its face, like 10 days, probably wasn't able to germinate in the presence of the h2o2, but once the h2o2 boke down it took over town. i dont think in that concentration the h2o2 actually "killed" it, it just prevented it from getting started for a little while.

on the other hand if you use some sort of bulk spawning method so that the myc has such a head start and quick colonization, you might be able to colonize it before the h2o2 breaks down... so more like mixing an equal part of colonized substrate to uncolonized substrate and crossing your fingers, you might pull it off.

oh, PS, it was a small tray full of BRF/verm mix (about as much as would fit into a half pint) treated with the H2O2 solution, and inoculated with a clean agar wedge. it grew some but didnt get very far and certainly not quick enough.


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Invisibledysphoria
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Re: Peroxide Substrate [Re: zeegos]
    #5650176 - 05/19/06 01:11 PM (17 years, 8 months ago)

you dont actually have to have a completely sterile environment.
you just have to have an excellent grasp of sterile technique.
the reason i would preach sterile environments to new growers is to cut down on the risk of contam, while they develop their technique. once thats done, it doesnt matter, a clean counter top in the middle of a dirty/ill kept house, with nothing more than butane torches and alcohol swipes for the needle, has proved to work for many people.

to answer at least one of your questions that i feel viable to answer, 10% or less h2o2 diluted in water has proven to not damage mycelium.
and while i dont know why, i just dont think adding peroxide to jars would work...probably couldnt/cant kill all of the bacteria/foreign spores in it before the h2o2 broke down into water. if it was that easy, we wouldnt be investing in pressure cookers or devising plans of fractional sterilization of brf cakes.


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InvisibleFungusMan
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Re: Peroxide Substrate [Re: dysphoria]
    #5650302 - 05/19/06 01:42 PM (17 years, 8 months ago)

Lol you answered your own damn question...
"hydrogen peroxide kills all bacterial spores apart from BIRTHED SPORES and LIVING MYCELIUM."

Peroxide kills spores...As the great Mencia once said..."dit dit duhhhhhhh!"


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Offlineskeletor
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Re: Peroxide Substrate [Re: FungusMan]
    #5650331 - 05/19/06 01:47 PM (17 years, 8 months ago)

well if you injected it with and LC you wouldn't have a problem with the spores dying. unfortunetly when you PC h202 it would break down. so there would be no benifit at all.


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Offlinezeegos
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Re: Peroxide Substrate [Re: skeletor]
    #5650671 - 05/19/06 03:35 PM (17 years, 8 months ago)

It seemed like a realistic idea but then again, like you said, we would have all adopted it by now if it were possible.
I may try it myself once my myco farm bag is fully colonised.
Its nice to see you had it going nicely for a while creamcorn, with just a 10:1 (does that mean 10:1 water:100% peroxide or 10:1
water:3% peroxide). if you used a really dilute solution and it lasted a while, i have 17.5% H2O2 i could dilute down to a 9 to 10% conc which may have enough peroxide molecules to totaly eliminate all bacterial spores. But if it wasa 10% solution you used, then i guess thats this theory out the window.


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Invisiblecreamcorn
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Re: Peroxide Substrate [Re: zeegos]
    #5650765 - 05/19/06 04:01 PM (17 years, 8 months ago)

yeah sorry i had my ratio numbers sorta backwards and confusing... 1 part 3% h2o2 to 10 parts h2o.

myc can tolerate h2o2, but it doesn't really like it. too much of the stuff will stall your myc just like it stalls the contams. it can also cause mutations, so be careful with the higher percentages.

for more info see this - you gotta buy his little ebooks to get the full story, but you'll see there he basically has a method for using peroxide to treat bulk substrates rather than sterilizing them, kind of like what i was getting at. you still need plenty of spawn so the bulk sub colonizes quickly, because once that h2o2 breaks down its obviously not doing you any good!


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Offlinezeegos
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Re: Peroxide Substrate [Re: creamcorn]
    #5650795 - 05/19/06 04:08 PM (17 years, 8 months ago)

yea i found his whole book posted on a site so i just got it from there :P
One other idea i have thought about is to do with cloaning.
When people colne shrooms, they slice off a top layre and take the fresh, unexposed tissue from underneath, open up the container and add into it.
I was wondering if a syringe could be used simply by injecting into a mushroom stem to get a core of tissue in the needle and inject into substrate jar.
Good idea??


Edited by zeegos (05/19/06 04:10 PM)


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Invisiblecreamcorn
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Re: Peroxide Substrate [Re: zeegos]
    #5650809 - 05/19/06 04:13 PM (17 years, 8 months ago)

not only a good idea but that's exactly how i do it :smile:

trap a little piece inside the syringe needle by poking right through the stem, shoot it out into a liquid culture, few days later you got cleanly cloned mycelium swimming around, and you can fill tons of syringes with the LC to shoot up whatever you want

it could certainly work with a substrate jar too, but the piece you get in the needle is really small, it'll have to land just right inside your substrate so its touching some "food" and i'd imagine it taking quite some time to really take off.  the lc way works well and reliably on the other hand, and the resulting liquid can be sprayed all over your sub so there's lots of contact points.


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Offlinezeegos
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Re: Peroxide Substrate [Re: creamcorn]
    #5650825 - 05/19/06 04:19 PM (17 years, 8 months ago)

haha, top notch.
sry im kinda noobie, what do you mean by you add the tissue to liquid culture? what liquid culture?

I just read up on what liquid culture was, wierd how i have never came across it b4. I knew spore syringes were full of liquid culture but never knew you could grow mycelium in liquid.
Kinda beats making a spore syringe, eh?


Edited by zeegos (05/19/06 04:27 PM)


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Invisiblecreamcorn
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Re: Peroxide Substrate [Re: zeegos]
    #5650846 - 05/19/06 04:26 PM (17 years, 8 months ago)

liquid culture is basically water, with a small amount of a simple sugar added for food.  usually done at a ratio of 96% water 4% sugars.  things like corn syrup, honey, glucose, malt extract, etc can be used.  mycelium will gladly grow within the liquid, and you can fill up syringes full of live mycelium bits suspended in the liquid and use that to inoculate.  its easy to make huge amounts, and stores for months in the fridge for later use.  theres about a zillion posts on here already about LC so i'll let you get to reading some of those and see. i promise it'll be the most useful thing you learn :smile:


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Offlinetrentallica
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Re: Peroxide Substrate [Re: zeegos]
    #5651297 - 05/19/06 06:56 PM (17 years, 8 months ago)

Quote:

zeegos said:
yea i found his whole book posted on a site so i just got it from there :P
One other idea i have thought about is to do with cloaning.
When people colne shrooms, they slice off a top layre and take the fresh, unexposed tissue from underneath, open up the container and add into it.
I was wondering if a syringe could be used simply by injecting into a mushroom stem to get a core of tissue in the needle and inject into substrate jar.
Good idea??




i like were your going with that. let me sugest that insted of using a needle to try and suck up a small flesh core use that same needle with a few cc's of h2o to inject into the shroom before pulling the plunger. i figure you should be able to pick up more shroom pieces that way.


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Offlinezeegos
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Re: Peroxide Substrate [Re: trentallica]
    #5657233 - 05/21/06 05:17 PM (17 years, 8 months ago)

Would it be ok to use golden syrup? Think thats all i got right now.
Once my myco bag is fully colonised im gonna inject some of my sterilized LC solution into the bag and suck it back up with some mycelium then back into the jar of LC solution to colonise. Got that from one of the 'poor man' teks.


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