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thedreammachine
Docter

Registered: 11/04/13
Posts: 14
Last seen: 1 year, 7 months
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Dead jars after liquid culture incoulation
#19087206 - 11/04/13 08:10 PM (10 years, 2 months ago) |
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Hey guys, first of all this is my first post on this wonderful site. Im a little puzzled on where i went wrong with this set up and procedure.
I have the puerto rico strain that i injected into a liquid culture and let it sit a week and a half in my closet. There was very slow myc growth and since this is my first time preforming an LC i thought maybe it was time to extract some and incoualted a couple jars with oak flower and vermiculite. I couldn't find any BRF, i thought i would not be that big of a deal its just nutrient for the MYC, right? Its been like 3 weeks now and the LC has a bunch of little white dots all up in the water.
So did i just pull the trigger to quick and didnt give time for the MYC to grow in the LC or is it the oak flower? Because there has been no activity and its been a week and a half with these cakes.
Thanks
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,810
Loc: Canada
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Re: Dead jars after liquid culture incoulation [Re: thedreammachine]
#19087285 - 11/04/13 08:30 PM (10 years, 2 months ago) |
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Its probably contamed I wouldn't trust a LC inoculated with anything but an agar wedge.
However spores can take up to 3 weeks to germinate in some cases. Patience is key in this hobby.
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TrentBoyett
Aspiring Mycologist



Registered: 11/29/12
Posts: 16,000
Loc: Kazakhstan
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Re: Dead jars after liquid culture incoulation [Re: Pastywhyte]
#19087737 - 11/04/13 10:25 PM (10 years, 2 months ago) |
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Idk what oak flower is, but you can grow mushrooms on a lot of stuff so idk if that's causing the problem or not.
You probably didn't give the mycelium enough time to colonize the lc.
LCs from spores are sketchy, always run a test jar before you use it.
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Kagenical
Stranger

Registered: 05/27/08
Posts: 128
Last seen: 10 years, 2 months
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Re: Dead jars after liquid culture incoulation [Re: TrentBoyett]
#19087784 - 11/04/13 10:36 PM (10 years, 2 months ago) |
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Most multispore LC's will take around a month to mature...
Many cloud within the first 2 weeks after inoculation, then clear up as the mycelium takes over the liquid and kills off any competing organisms. In my experience, many LC's will contaminate within the first 1-3 weeks, then after they mature, will make many successful inoculations.
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TrentBoyett
Aspiring Mycologist



Registered: 11/29/12
Posts: 16,000
Loc: Kazakhstan
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Re: Dead jars after liquid culture incoulation [Re: Kagenical]
#19087813 - 11/04/13 10:41 PM (10 years, 2 months ago) |
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No, if your LC contams throw it out, the contam will still be in the LC even if it doesn't appear to be, it's hard to tell if a LC is contammed.
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Kagenical
Stranger

Registered: 05/27/08
Posts: 128
Last seen: 10 years, 2 months
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Re: Dead jars after liquid culture incoulation [Re: TrentBoyett]
#19087866 - 11/04/13 10:56 PM (10 years, 2 months ago) |
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I've not found that to be true.
I've done many MS LCs and all of them clouded substantially after inoculation, only to turn crystal clear about 4-5 weeks after mycelium starts to form. I always shoot LC to agar first to ensure it's viability and check for contams. Many of my LC's have produced bacterial contamination within the first few weeks then cleaned themselves up.
I used a master culture, which behaved this way for almost an entire year and produced no contaminated rye jars. Literally, a 100% success rate.
I'm not saying this is always the case, but to assume a spore syringe is 100% free of any genetics but mushroom spores is asinine. There will always be an endospore or two in it, these will germinate much faster than mushroom spores, and will typically be out-competed by the spores as time goes on. If you sterilized your culture properly, many will clean up if your syringes are sufficiently clean.
Though I will admit in the OP's case, he's likely FUBARd.
Edited by Kagenical (11/04/13 10:57 PM)
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TrentBoyett
Aspiring Mycologist



Registered: 11/29/12
Posts: 16,000
Loc: Kazakhstan
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Re: Dead jars after liquid culture incoulation [Re: Kagenical]
#19087894 - 11/04/13 11:04 PM (10 years, 2 months ago) |
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You're probably not actually seeing a contam in your LC, something else is making it cloudy, like the spore solution or something.
Then you're probably getting contams on the test because not enough of the lc is colonized with mushroom mycelium so it is more open to contains.
Just my opinion.
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