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InvisibleQuiescence
Intriguer

Registered: 04/25/05
Posts: 315
Liquid Culture Contamination Attempt and Failure.. in a good way?
    #4331137 - 06/24/05 01:17 AM (18 years, 9 months ago)

So how do you know when a liquid culture is contaminated? I always heard.. "you'll know it when you see it" .. well I have never had a contaminated culture.. to my knowledge at least. Never had a problem where all the jars I knock up die on me.. so I don't think my LC's have ever been a problem.

SO, I wanted to know how you can tell when an LC is actually contaminated. To do this, I scraped up a big chunk of spores, with my unwashed finger, from around the fan blade that I use in my fruiting chamber. I stir a big chunk into 3 LC's that I made using tap water and honey that I did nothing to. Just poured it in, open air. Shook the jars and incubated them.

Nearly two weeks later and all three have significant growth. Big gooey balls of mycelium sperm. But they all look completely normal. There are some black spots.. but I am pretty certain they are ungerminated spores or other stuff that was on the fan... they are just small black dots like you would normally see if you had a big ball of spores.

So do I just need to wait longer for contamination to appear? This seems like it would be long enough. I did all I could think of to make them 'wrong'...

What's up?

~Quiescence~


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God raised up the fellow who was relaxed in the company of tax collectors, prostitutes and other outsiders, and went on to say that many of them would be welcomed to God?s banquet table long before those self-righteous people on display at church.

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Offlinelepiota
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Registered: 10/27/04
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Re: Liquid Culture Contamination Attempt and Failure.. in a good way? [Re: Quiescence]
    #4331154 - 06/24/05 01:26 AM (18 years, 9 months ago)

i had some contaminated LCs: they smelled bad.


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Offlinecaimen
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Registered: 06/22/04
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Re: Liquid Culture Contamination Attempt and Failure.. in a good way? [Re: Quiescence]
    #4331158 - 06/24/05 01:29 AM (18 years, 9 months ago)

I saw a T.V. show that said Honey will slow or provent bacteria and mold growth but Im not 100% sure.


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Invisibleagar
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Re: Liquid Culture Contamination Attempt and Failure.. in a good way? [Re: Quiescence]
    #4331163 - 06/24/05 01:34 AM (18 years, 9 months ago)

Leave them alone & don't agitate them. Sooner or later - you will see light buff blue growing on the surface of the LC, or some color other than white Myc.


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Offlinetracerz
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Registered: 05/08/05
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Loc: wa forest
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Re: Liquid Culture Contamination Attempt and Failure.. in a good way? [Re: lepiota]
    #4331173 - 06/24/05 01:36 AM (18 years, 9 months ago)

your right about the black dots with bacteria in karo the water turns slightly yellow or yellower in honey the most common lc problem.and you must have a clean finger there lol.

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InvisibleQuiescence
Intriguer

Registered: 04/25/05
Posts: 315
Re: Liquid Culture Contamination Attempt and Failure.. in a good way? [Re: caimen]
    #4331177 - 06/24/05 01:38 AM (18 years, 9 months ago)

lepiota, you mean a bad smell like when WBS sits in water too long? Similar yea? These guys have no smell at all..

Caimen, So You think these could NOT be contaminated? Can honey be that magical? Then why is everyone so anal about PC'ing LC's? If I can make an LC from spores I pick up with my fingers.. use tap water.. and intentionally stir with a dirty finger to get nasty stuff in thw water.. and Have it all turn out fine....

I guess my question is. How long before you Normally see a contaminate? What will it look like.. growth On the myc. Seperate growth? What contams usually affect LC's? Who has had a contaminated LC's .. and known it by looking at them?

~Quiescence~


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God raised up the fellow who was relaxed in the company of tax collectors, prostitutes and other outsiders, and went on to say that many of them would be welcomed to God?s banquet table long before those self-righteous people on display at church.

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InvisibleQuiescence
Intriguer

Registered: 04/25/05
Posts: 315
Re: Liquid Culture Contamination Attempt and Failure.. in a good way? [Re: agar]
    #4331180 - 06/24/05 01:39 AM (18 years, 9 months ago)

Awesome. Thanks agar..

So watch for stuff on the surface. Will do.

Thanks man.

~Quiescence~


--------------------
God raised up the fellow who was relaxed in the company of tax collectors, prostitutes and other outsiders, and went on to say that many of them would be welcomed to God?s banquet table long before those self-righteous people on display at church.

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Offlinelepiota
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Re: Liquid Culture Contamination Attempt and Failure.. in a good way? [Re: Quiescence]
    #4331322 - 06/24/05 03:11 AM (18 years, 9 months ago)

bed smell like when the wbs is contaminated by bacteria and looks jelly and smells like something sweet and rotten.
actually they were not liquid cultures but failed attempts of agar cultures in which water entered during sterilization and the polifill filter was completely wet with water+nutes.


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Invisibledeanofmean
mycophagous

Registered: 12/06/02
Posts: 2,017
Loc: PNW
Re: Liquid Culture Contamination Attempt and Failure.. in a good way? [Re: Quiescence]
    #4331354 - 06/24/05 03:58 AM (18 years, 9 months ago)

I'm calling BULLSHIT if you did the procedure that you claim, and haven't noticed any contaminates after two weeks, you must be in a sterile bubble, or in outer space.  :evil:


edited 4 spelling

Edited by deanofmean (06/24/05 04:31 AM)

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Offlinelordoftheshroomz
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Re: Liquid Culture Contamination Attempt and Failure.. in a good way? [Re: deanofmean]
    #4331469 - 06/24/05 06:45 AM (18 years, 9 months ago)

nice avatar lepiota


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OfflineIGnosticAbhorI
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Re: Liquid Culture Contamination Attempt and Failure.. in a good way? [Re: lordoftheshroomz]
    #4331517 - 06/24/05 07:26 AM (18 years, 9 months ago)

Quote:

Big gooey balls of mycelium sperm.




:rotfl:

Mine ALWAYS look white...

Green starts out white and doesn't really get green until it starts to produce spores...and i'm sure that in a liquid environment it'd be nearly impossible for it to generate.

My point - Mine never have a color.

They either smell...or I ca'nt tell lol...

I can always tell 2-3 days after knock up.

I've seen Tric. so much in my jars (mainly from lc's), that I can spot tric. early in it's whitey tidey stage :cool:.

-Gnostic

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Offlinescatmanrav
Brainy Smurf

Registered: 05/08/04
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Re: Liquid Culture Contamination Attempt and Failure.. in a good way? [Re: IGnosticAbhorI]
    #4331679 - 06/24/05 09:19 AM (18 years, 9 months ago)

Quote:

IGnosticAbhorI said:
Quote:

Big gooey balls of mycelium sperm.




:rotfl:

Mine ALWAYS look white...

Green starts out white and doesn't really get green until it starts to produce spores...and i'm sure that in a liquid environment it'd be nearly impossible for it to generate.

My point - Mine never have a color.

They either smell...or I ca'nt tell lol...

I can always tell 2-3 days after knock up.

I've seen Tric. so much in my jars (mainly from lc's), that I can spot tric. early in it's whitey tidey stage :cool:.

-Gnostic




^^^^yep....only after alot of time do I get color, if at all when I have contams.

I also believe, his procedure is POSSIBLE to get without contams...but very unlikly..


--------------------
"life is like a drop of rain getting closer and closer to falling into a lake, and then when you hit the lake there is no more rain drop, only the lake."

Growing with bags, start to finish (including my new grain and substrate prep)
Anyone looking to start bulk tubs/mono tubs/shotgun hybrids? Good tubs to use..
How I do grain (old still good tips)
Turn your closet into a fruiting chamber
Casing layer colonization and overlay

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InvisibleQuiescence
Intriguer

Registered: 04/25/05
Posts: 315
Re: Liquid Culture Contamination Attempt and Failure.. in a good way? [Re: scatmanrav]
    #4332309 - 06/24/05 12:04 PM (18 years, 9 months ago)

So replies for everyone.
Deanofmean: I shit you not. It wasn't two weeks yet.. like 11-12 days.. I can't remember when I did them exactly.

Iggy: That would make sense. How different does trich look? They are in half-pinters .. the ones with the wavy lines so it is hard to see well. Plus its honey water so it is yellow. Quite possible I can't tell yet...

scatmanrav: Hmm possible to not be contaminated? Heh.. I really doubt it... I mean possible I guess... but those spores had been in the fan for over a week... plus.. the finger... I can't imagine they don't have something in there. I just think I can't tell yet...

So basically I am leaving them alone in a seperate place. I will check in a week or so. See if stuff is floating like agar said. Check for any new smells.. and maybe some color.

Thanks for the replies.

~Quiescence~


--------------------
God raised up the fellow who was relaxed in the company of tax collectors, prostitutes and other outsiders, and went on to say that many of them would be welcomed to God?s banquet table long before those self-righteous people on display at church.

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Offlinelepiota
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Registered: 10/27/04
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Re: Liquid Culture Contamination Attempt and Failure.. in a good way? [Re: IGnosticAbhorI]
    #4332314 - 06/24/05 12:05 PM (18 years, 9 months ago)

Quote:

IGnosticAbhorI said:
Green starts out white and doesn't really get green until it starts to produce spores...and i'm sure that in a liquid environment it'd be nearly impossible for it to generate.
My point - Mine never have a color.
They either smell...or I ca'nt tell lol...
I can always tell 2-3 days after knock up.





uhmmm... now listen what happened to me: inoculated 8 PF jars with a liquid culture, it was my 1st liquid culture so i'm usure about the correctness of the procedure, anyway, they colonized completely in an extra short time if compared to spore syringes (1 week). they were completely white when i birthed them, and i mean white. after one week from birth they all got contamined with green mold.
do you think it was something inside my LC, or it would have shown up during incubation, and the problem is in my PMP?


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Offlinescatmanrav
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Registered: 05/08/04
Posts: 11,483
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Re: Liquid Culture Contamination Attempt and Failure.. in a good way? [Re: Quiescence]
    #4332384 - 06/24/05 12:23 PM (18 years, 9 months ago)

Quote:

Quiescence said:
scatmanrav: Hmm possible to not be contaminated? Heh.. I really doubt it... I mean possible I guess... but those spores had been in the fan for over a week... plus.. the finger... I can't imagine they don't have something in there. I just think I can't tell yet...

~Quiescence~




Someone claimed to do eactly what you did, but using a print instead of the fan blade. Using his finger, into the LC, open air in his dirty bedroom. And he says they turned out fine...I could believe this.

I could also believe that in my chamber, I keep contams out, so if you havnt had a contam in your chamber, its quite likly that there arent to many mold spores in there, and that the ones that were there, dont land on the spot where you scrape..and even if a few spores are right there, its possible with that many cubie spores to fight it off. I just know that I've gone 7 months, without cleaning my fan in a closet twice the size of a martha...and contams were never a problem. If there were that many spores floating around in my chamber I think I'd have more of a contam problem. As it is, I really never get contamed casings...I have yet to see mold growing on poo, or a casing with poo..I dont open jars when they get contamed or clean them out, just trash the whole jar, everythings kept very clean, HEPA rnning outside the closet all the time...and there were spores everwhere in the closet, and fan, and alot germinated on the moist plastic walls...never molds though.

So yeah, I think its possible to happen, a few times out of many trials...but unlikly...

Green mold in LC looks slightly different, pretty close..maybe a little more like cotton balls..sometimes I cant see it at all though and it turns out contamed. And bacteria is really hard to see sometimes.


--------------------
"life is like a drop of rain getting closer and closer to falling into a lake, and then when you hit the lake there is no more rain drop, only the lake."

Growing with bags, start to finish (including my new grain and substrate prep)
Anyone looking to start bulk tubs/mono tubs/shotgun hybrids? Good tubs to use..
How I do grain (old still good tips)
Turn your closet into a fruiting chamber
Casing layer colonization and overlay

Extras: Filter Print Post Top
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