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OfflineHayfork
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Ur Opinions on My Contains Plz
    #26548939 - 03/21/20 01:29 PM (3 years, 10 months ago)

How's it going gang,

Dusted off my gear and resumed my small BRF setup.  Had some 1-year-old liquid culture that I added some sugar to in hopes of reviving.  A week later injected the LC into some BRF jars. Jars were off to a really good start but all 16 of them started to develop green mold right below the top verm  layer.  I've never encountered this pattern of green mold before in my jars during incubation.  I'm getting ready to toss the LC and make a couple new jars of LC.  Can you folks think of something that might be causing this type of contamination?  I would hate to make the same mistake twice. 

I'm well aware of the risks of LC but have had good luck in the past, not set up for agar at the moment.


Thanks for your help. Pictures attached.





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Offlinehazyhorse
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Re: Ur Opinions on My Contains Plz [Re: Hayfork]
    #26549003 - 03/21/20 02:05 PM (3 years, 10 months ago)

https://www.shroomery.org/forums/showflat.php/Number/26096469/page/1

i had something similar happen when i used a LC i didn’t clean up on agar first. i don’t know what it was & never got any answers. but i have gotten that kind of contamination before. i didn’t end up fruiting the jars


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InvisiblefahtsterM
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Re: Ur Opinions on My Contains Plz [Re: hazyhorse] * 1
    #26549032 - 03/21/20 02:26 PM (3 years, 10 months ago)

Old LC.. might be best to start from spore syringe to pf jars.  Might be penicillin in those jars... Probably got in when you added the sugar

Faht


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OfflineHayfork
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Re: Ur Opinions on My Contains Plz [Re: fahtster]
    #26549143 - 03/21/20 03:33 PM (3 years, 10 months ago)

I have no problem starting over.  I've goty pretty good at making reliable LC.  My main concern was maybe a flaw in my sterilization or inoculation procedures that might be obvious to you veterans out there.

I've had plenty of contams after birthing jars.  Very rarely in jars during incubation and never like this instance, exactly where the BRF meets top 3/8" thick layer of verm on every single jar.


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InvisiblefahtsterM
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Re: Ur Opinions on My Contains Plz [Re: Hayfork]
    #26549161 - 03/21/20 03:42 PM (3 years, 10 months ago)

Yeah it’s hard to say.. with both sterilization and inoculation you would think that the top isn’t the only place you would contamination.. if it were your iniculant, your probably see it all over and the top of the jars is where the steam enters first during sterilization, although, it could have worked it’s way out from the center and there’s contamination behind the myc that’s showing.  I’m assuming you used a pc and you’re using a dry verm barrier?

The year old LC seems suspect to me and adding nutes to it after the fact.  How exactly did you go about that?

Faht


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OfflineHayfork
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Re: Ur Opinions on My Contains Plz [Re: fahtster]
    #26550571 - 03/22/20 09:47 AM (3 years, 10 months ago)

Yes PC my jars for 60 minutes.

I added nutes to old LC by dissolving unrefined rock sugar in water and bringing to boil in microwave.  Sterilized syringe by cycling alcohol then H2O2 through it. 

Injected nutes through self healing port while spraying everything with copius amounts of alcohol.  Didn't see any vacuum while removing syringe.

I made 4 new LC last night.  Will test in a couple weeks.  If I encounter same pattern of contams I'll bother you guys for more help. 

🤞🤞🤞 Thanks for all your inputs.

On a positive side note.  I already had tons of alcohol and H2O2 on my shelves when the stores ran out😜


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InvisiblefahtsterM
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Re: Ur Opinions on My Contains Plz [Re: Hayfork]
    #26550730 - 03/22/20 11:05 AM (3 years, 10 months ago)

Well, I think the cycling of alcohol and h2o2 through the syringe was the culprit... that and using the microwave.  Probably would have been better off just sterilizing the syring in the pc and using the lc as it was if it was alive enough to still grow.  But if you were to add the sugar water to it, it would be better to dissolve the sugar in water on the stove, bring to a boil for 15 minutes and then cycle that water (while still boiling) through the syringe a few times and on the last time, keeping the water in the syringe and let cool.

Or just use the pc to sterilize the syringe with sugar water in it

You probably only sanitized the syringe with the alcohol and h2o2. The microwave can be inconsistent.

Faht


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