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Invisiblefngbronco
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F*** bacteria! Possible solution? * 1
    #14265846 - 04/10/11 07:45 AM (12 years, 10 months ago)

I've gone through and revamped everything! Synthetic filter discs for GE on all my jars, Lysol, 90% alcohol, 90-120 minute PC times from 15-20 psi, super sterile glove box technique (so I think). My agar dishes are cleaner for sure except some small amounts of bac every now and then and my grains sit perfectly until I inoculated them with an LC. I know 100% the bac came in through the lc and it sucks. My first lc expansion and it is growing great minus it's little passenger. My jars got about 40-60% before being overrun. I was curious if I could add an antibiotic (gentamiacin) to the LC to clear up the bac. Of course my jars are already dumped into compost, I was just curious if it'd work for the lc. All that I've read are ?s with suggestions, but no real answers or follow up. People have recommended against it for grains or as an initial additive though.

Has anyone tried it or had any experience? This jar isn't the end of the world to me but it saddens me that it may have to go. I'd hate to waste the gent, or create some super strain bac colony by just adding enough it has the anti-venom affect on the colony rather than killing it off.


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I challenge you to challenge yourself more! When you feel complacent and ready to hang it up, challenge yourself to get over it! If you fail, don't look at it as you didn't succeed, look at it as you would a rock face you're trying to climb. Stand back, wayyyy back, and look at it and plot another path. If you can't find one, shuffle down the way a little, a little change of scenery or a view from a different angle may give you the insight you need.

Anything I state is relayed information from a friend of a friend and should be viewed as completely fictitious. I do not partake in any illegal or grey-area-of-the-law activities, but do have lots of friends who may or may not.    -fngbronco

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Offlinex7x_x7x
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Re: F*** bacteria! Possible solution? [Re: fngbronco]
    #14266822 - 04/10/11 12:21 PM (12 years, 10 months ago)

the problem could be the grain. soak you the grain 12 hs before PC? if not, soak for 6-24 hs in a well filled pot, simmer, pour and leave it cool for 15 minutes, the fill the jars and PC.


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koh samui and oak ridge are my favourite strains

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Invisiblefngbronco
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Re: F*** bacteria! Possible solution? [Re: x7x_x7x]
    #14266842 - 04/10/11 12:27 PM (12 years, 10 months ago)

I put the grains outside and let them dry (exterior) then loaded and pc'd. I kept one control jar to make sure that my grains were good and my control is still clean and purty. I put 2.5mg in the LC and spun it. Hopefully it's enough I hear the gent is pretty strong stuff.


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I challenge you to challenge yourself more! When you feel complacent and ready to hang it up, challenge yourself to get over it! If you fail, don't look at it as you didn't succeed, look at it as you would a rock face you're trying to climb. Stand back, wayyyy back, and look at it and plot another path. If you can't find one, shuffle down the way a little, a little change of scenery or a view from a different angle may give you the insight you need.

Anything I state is relayed information from a friend of a friend and should be viewed as completely fictitious. I do not partake in any illegal or grey-area-of-the-law activities, but do have lots of friends who may or may not.    -fngbronco

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OfflineBrennus
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Re: F*** bacteria! Possible solution? [Re: fngbronco]
    #14267059 - 04/10/11 01:16 PM (12 years, 10 months ago)

This would've been a better topic in the Contamination Forum.

Your problem sounds like it's based in using a sugar-based liquid culture. These types of LC's are an amazingly great way to suffer massive contamination rates for a home-grower. Stick with spores or inoculating with agar wedges that you know are clean and see if you still have problems, especially if it's the only vector of contamination you haven't accounted for. Adding in antibiotics to a batch of LC isn't going to solve anything, either.

Additionally, "Grain LC" or mycelium-laden water is likely your best bet if you really want to get the cleanest bang for your buck. After shaking up a clean grain master, mix in sterile water and swish it around the jar. Aspirate into a syringe.

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Invisiblefngbronco
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Re: F*** bacteria! Possible solution? [Re: Brennus]
    #14267138 - 04/10/11 01:33 PM (12 years, 10 months ago)

Good point. My luck with agar has been shit at best unless dealing with wedges or tissue. I had a supplier's lc and tried expanding it. I'll definitely try the grain lc once I get a grain jar to survive lol. That's for the help.


Also can I have a mod move this to contam?


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I challenge you to challenge yourself more! When you feel complacent and ready to hang it up, challenge yourself to get over it! If you fail, don't look at it as you didn't succeed, look at it as you would a rock face you're trying to climb. Stand back, wayyyy back, and look at it and plot another path. If you can't find one, shuffle down the way a little, a little change of scenery or a view from a different angle may give you the insight you need.

Anything I state is relayed information from a friend of a friend and should be viewed as completely fictitious. I do not partake in any illegal or grey-area-of-the-law activities, but do have lots of friends who may or may not.    -fngbronco

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Offlinetotal
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Re: F*** bacteria! Possible solution? [Re: fngbronco]
    #14267439 - 04/10/11 02:46 PM (12 years, 10 months ago)

Did you get the gent sulfate from me? :shrug:
I dont see any reason why you couldnt add it to a LC to help prevent bacterial contams...
It comes "unsterile" and should be added prior to pressure cooking though...:super:

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Offlinex7x_x7x
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Re: F*** bacteria! Possible solution? [Re: fngbronco]
    #14267459 - 04/10/11 02:50 PM (12 years, 10 months ago)

when you put the grain outside, could become contaminated again with new bacterial spores.

from my point of view, are no other reason that justify the contamination. i been working with honey LC for years with an extremely low contam rate.

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Offlinex7x_x7x
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Re: F*** bacteria! Possible solution? [Re: x7x_x7x]
    #14267471 - 04/10/11 02:52 PM (12 years, 10 months ago)

which mushroom are you growing? start the culture with spores o mycelium?

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Invisiblefngbronco
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Re: F*** bacteria! Possible solution? [Re: x7x_x7x]
    #14267571 - 04/10/11 03:16 PM (12 years, 10 months ago)

Yeah the gs is from you total. I'm not worried about it's sterility as the culture is already buggy.

The lc was started with some oyster lc from a sponsor. It grew out well, but the transfer to more lc bugged. It's clear and there's tons of good growth but some cloudiness recently above the myc level. I'll post a pic soon as I get home. The jars I knocked up with it showed really decent growth for a little bit but then got wet looking. I've tossed them already though.


--------------------
I challenge you to challenge yourself more! When you feel complacent and ready to hang it up, challenge yourself to get over it! If you fail, don't look at it as you didn't succeed, look at it as you would a rock face you're trying to climb. Stand back, wayyyy back, and look at it and plot another path. If you can't find one, shuffle down the way a little, a little change of scenery or a view from a different angle may give you the insight you need.

Anything I state is relayed information from a friend of a friend and should be viewed as completely fictitious. I do not partake in any illegal or grey-area-of-the-law activities, but do have lots of friends who may or may not.    -fngbronco

Pill Divider Agar Tek

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Offlinex7x_x7x
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Re: F*** bacteria! Possible solution? [Re: fngbronco]
    #14267588 - 04/10/11 03:21 PM (12 years, 10 months ago)

some water is normal

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Invisiblefngbronco
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Re: F*** bacteria! Possible solution? [Re: x7x_x7x]
    #14267616 - 04/10/11 03:29 PM (12 years, 10 months ago)

Oh the jars are definitely bacteria, I meant they looked wet like bacteria took over the whole jars.


--------------------
I challenge you to challenge yourself more! When you feel complacent and ready to hang it up, challenge yourself to get over it! If you fail, don't look at it as you didn't succeed, look at it as you would a rock face you're trying to climb. Stand back, wayyyy back, and look at it and plot another path. If you can't find one, shuffle down the way a little, a little change of scenery or a view from a different angle may give you the insight you need.

Anything I state is relayed information from a friend of a friend and should be viewed as completely fictitious. I do not partake in any illegal or grey-area-of-the-law activities, but do have lots of friends who may or may not.    -fngbronco

Pill Divider Agar Tek

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Offlinex7x_x7x
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Re: F*** bacteria! Possible solution? [Re: fngbronco]
    #14267841 - 04/10/11 04:34 PM (12 years, 10 months ago)

ugh

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OfflineRogerRabbitM
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Re: F*** bacteria! Possible solution? (moved) [Re: fngbronco]
    #14269726 - 04/10/11 10:19 PM (12 years, 10 months ago)

This thread was moved from Advanced Mycology.

Reason:
If you have a clean culture on agar, simply use it to inoculate a grain master, and then expand further by grain to grain transfers.  I think you'll have better luck with that in a glovebox than LC, and it's a heck of a lot faster. Spray the inside of your glovebox with water just before working.  The particles in the air will be drawn to the water droplets and contained.
RR

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OfflineMedici_head
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Re: F*** bacteria! Possible solution? (moved) [Re: RogerRabbit]
    #14274285 - 04/11/11 07:02 PM (12 years, 10 months ago)

check out sabouraud dextrose agar, it's used in the health industry for promoting only mycological growth and suppressing bacterial growth. a small bit will go a long way and especially good for home-growers.  what's your agar composition?

medici


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Y o u ' r e  a l l  G o d s ;  Y o u ' r e  a l l  G o d s ;  Y o u ' r e  a l l  2  m i l l i o n  y e a r s  o l d . ~T. Leary

Edited by Medici_head (04/11/11 07:26 PM)

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