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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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Bacteria Problem
    #7275507 -

Hey folks!

I seem to be having some bacteria problems with my BRF cakes. At every inoculation site on every jar I've knocked up has a bacteria blotch. This is leading me to believe that I am introducing the bacteria while innoculating. The strange thing is that I flame sterilize the spore syringe prior to innoculating every jar. Let me give you a run down of my procedure:

I am following the pf-tek using half-pint mason jars. The lids contain the typical 4 inoculation points and they are covered with micropore tape. I place a foil cover over the jars and PC at 15 psi for 45 minutes. When finished, I leave the jars in the PC for 12 hours or so to cool. When I'm ready to inoculate I open the lid on the PC and remove a jar. I flame the needle red hot, allow some spore solution to shoot out the end, then quickly remove the foil cover and inoculate the jars. I don't remove the micropore tape, I just puncture right through it. After that it's into the incubator at 80'F.

Is there a better method to what I am doing? Can you spot how I could be introducing bacteria into the jar? Any help would be much appreciated.

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Re: Bacteria Problem [Re: leatherlung]
    #7275555 -

invest in some RTV high temp silicone, then place a good dab over every innoc. hole. This gives you a self sealing innoc. port. It's really easy for just one little contam spore to get in.


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c        l        i      c        k        m        e

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Re: Bacteria Problem [Re: aspore]
    #7275567 -

Won't this plug the inoculation points, preventing any FAE though?

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Re: Bacteria Problem [Re: leatherlung]
    #7275754 -

The above makes it sound like your inoculate is contaminated.
RR


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Download Let's Grow Mushrooms



semper in excretia sumus solim profundum variat

"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
Thomas Edison

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Re: Bacteria Problem [Re: RogerRabbit]
    #7276273 -

Hmmn, definitely a possiblity. That's what sucks, there are so many possibilities! I really don't want to accuse the vendor of providing a contaminated product; there is likely a flaw in my procedure somewhere. But then how would you ever know? And how responsive would a vendor be to such claims?

Spores101.com seem to be a reputable company. Has anyone else had problems with these guys?

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Re: Bacteria Problem [Re: leatherlung]
    #7276293 -

try to also alcohol swab the needle after you flame it. i flame it until it's red hot for each jar and i rub an alcohol swab over it before each hole. i use a new alcohol swab for each jar.

after you flame sterilize the needle, it's fine, but tiny mold spores and whatnot in the air can land on it inbetween each hole.

but id still go with what RR said due to the fact that every hole has a blotch.


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http://www.doctorfungus.org

http://www.mushroomexpert.com/

http://www.textbookofbacteriology.net

http://bugs.bio.usyd.edu.au/learning/resources/Mycology/index.html

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Re: Bacteria Problem [Re: LayYouIn]
    #7276482 -

Yeah... contaminated inoculant is the last thing most people seem to suspect, yet it's usually the first culprit.

I am fully aware this isn't exactly ideal procedure (that's a nod and a wink to you, RR, before I'm yelled at :smile:), but if you are entirely reliant on this syringe to continue your work while you wait for a replacement, you can try and form an LC with it, then give it a few CCs of peroxide. I did a comparison run with an LC I suspected to be contaminated, knocked up five without peroxide, and seven with. All seven with peroxide came out clean, and all five without it came up chock full of metabolites (though several seem to be showing promise of pulling through despite the infection).

Now, you can't just add peroxide to the spore syringe, because your spores haven't germinated, and they'll be killed. But, since mushroom mycelium tends to survive peroxide exposure while bacteria are easily killed by it, it can be used to selectively clean up bad inoculant. Since you're not working with a contaminated LC, I don't know how well it will work; your spores might not germinate and colonize the LC if the bacteria have too big of a head start. But there's only one way to find out!

Note that this won't eradicate many molds, but it looks like you're not experiencing any.


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Re: Bacteria Problem [Re: figgusfiddus]
    #7276561 -

I've been reading your opinions on incubators as well figgus and I think I may just scrap my incubator as well. It isn't helping the problem any... At least the blotch is holding to just the inoculation sites and not spreading. I didn't use too much spore solution per jar, just the typical one to two CCs. However I am out of solution and waiting on another syringe. I think I am going to go the LC route this time around. The faster colonization times sounds absolutley worth it, not to mention more medium to work with.

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Re: Bacteria Problem [Re: leatherlung]
    #7276590 -

Yeah... LCs are great, but use your spore solution sparingly when making them. It's not that unusual for LCs to go bacterial (which is NOT the end of the world if you've got aggressive mycelium, though it's still not good), so you should always have a backup plan. I would never just rely on LCs for months on end; I always want a master print to come back to.


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Re: Bacteria Problem [Re: figgusfiddus]
    #7276915 -

How do you know when the LC is contaminated with bacteria? What are the tell-tale signs?

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Re: Bacteria Problem [Re: leatherlung]
    #7276937 -

You don't know until you inoculate with it. That's why it's kind of a bad technique for a first-timer.


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Re: Bacteria Problem [Re: figgusfiddus]
    #7276993 -

you could try having ur oven turned on with the oven door open and work in the hot air updraft, this may help prevent contams landing on ur stuff, along with disfecting the entire work area and even entire room.

Bacteria is everywhere, as i found out when i did not have enough verm/bfr mix for my second jar so i tinfoiled it and left it in the cupboard (without sterilising it) thinking i'll make up some more in a few days and pc it. just took it out 2day remember, damn thats gonna be contaminated, quick look and couldnt see anything then a smelled,. damn that thing stinks, gonna have to PC it to kill all the bacteria then throw it out.


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PF TEK - writeup by EvilMushroom666
Lets Grow Mushrooms - RogerRabbit & RoadKills website with sample videos plus the full PF TEK video series. Alot of great information - BUY THE DVD
Cakes can and will pin! - So you think cakes suck for pins. Your wrong
Franks Simple Coir/Verm Tek
Franks Proper Pasturisation Tek
Franks Spawning To Bulk - Monotub
Professor Pinheads RTV Injection Port Tek
Foo Mans No Soak WBS Prep Tek

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Re: Bacteria Problem [Re: leatherlung]
    #7277588 -

leatherlung said:
I've been reading your opinions on incubators as well figgus and I think I may just scrap my incubator as well.


if you've already made it, you might as well keep it. it'll help a little if you live in a cold place. or you could just use it as a monotub or FC.


--------------------
http://www.doctorfungus.org

http://www.mushroomexpert.com/

http://www.textbookofbacteriology.net

http://bugs.bio.usyd.edu.au/learning/resources/Mycology/index.html

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Re: Bacteria Problem [Re: veda_sticks]
    #7280017 -

veda_sticks said:
you could try having ur oven turned on with the oven door open and work in the hot air updraft, this may help prevent contams landing on ur stuff, along with disfecting the entire work area and even entire room.

Bacteria is everywhere, as i found out when i did not have enough verm/bfr mix for my second jar so i tinfoiled it and left it in the cupboard (without sterilising it) thinking i'll make up some more in a few days and pc it. just took it out 2day remember, damn thats gonna be contaminated, quick look and couldnt see anything then a smelled,. damn that thing stinks, gonna have to PC it to kill all the bacteria then throw it out.


Ew. Oven tek is supposed to be pretty rotten. People have made it work, but I strongly suspect it works on par with open-air or maybe a bit worse.


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