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Offlinellamaboy
the weasel thatsnagged the bee

Registered: 11/08/03
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Question about agar contam *pic*
    #2215265 - 01/02/04 05:18 PM (12 years, 11 months ago)

Okay, first off, this is an agar plate made from pda. the plate and everything was made under sterile conditions. i couldn't use a standard spore print to make this plate, as no one had the strain that i wanted in a plate, all anyone had was a syringe. so, i used the syringe. i dropped about two drops of water onto the plate and it's been incubating for about 2 days now. my question is this: what the fuck is that shit growing over it. it's directly where the water was placed and then spread to. i'm thinking that the standing water ontop of the agar has done something to the agar, but i'm not sure. i just want to know what you people think. i'm certain that those little white dots that you see there are soon too be colonies of mycelium, but what the hell is that green(don't know about the true colour of it, i can't really see colour that well.) shit that's all over the plate now? if anyone has seen anything like this before, please help me out...every single plate has this shit one it, and it's only where the water was.


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Offlinellamaboy
the weasel thatsnagged the bee

Registered: 11/08/03
Posts: 563
Loc: Portland PNW
Last seen: 7 years, 1 month
Re: Question about agar contam *pic* [Re: llamaboy]
    #2215266 - 01/02/04 05:19 PM (12 years, 11 months ago)

heh, so i just remembered that there's a contamination forum *argh* maybe it can sit here for a while...if it should be moved, move it for me would ya.


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OfflineGranola
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Re: Question about agar contam *pic* [Re: llamaboy]
    #2215561 - 01/02/04 08:08 PM (12 years, 11 months ago)

Mmmm, bacteria. looks like thoses little guys didnt stand a chance, next time, shake the hell out of the syringe, stand it on it's plunger and allow it to rest for several hours, push the syringe down the plunger and evacuate some of the water, shake it again and flame the needle, red hot all the way down, push about 1/5cc out and bring it to the agar, start another drop and drag it across the surface, push a couple drops and repeat the push & drag. try not to leave a full drop, the droplets that do remain will evaporate quickly.


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Offlinellamaboy
the weasel thatsnagged the bee

Registered: 11/08/03
Posts: 563
Loc: Portland PNW
Last seen: 7 years, 1 month
Re: Question about agar contam *pic* [Re: Granola]
    #2216275 - 01/03/04 06:27 AM (12 years, 11 months ago)

so, it is bacteria? it's not possible that it's some sort of reaction that the water had with the PDA? i mean all of the agar plates have this on it, right where the water was. if it were bacteria, would that mean that it was in the syringe? i followed all sterile techniques.


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InvisibleG a n j a
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Re: Question about agar contam *pic* [Re: llamaboy]
    #2216401 - 01/03/04 10:46 AM (12 years, 11 months ago)

Looks like a slime mould, or bacteria to me.
At the best a yeast infection, did you use yeast in the agar?


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OfflineSuntzu
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Re: Question about agar contam *pic* [Re: llamaboy]
    #2216698 - 01/03/04 03:04 PM (12 years, 11 months ago)

That is most definitely a mixed bacterial pool. At least three; Bacillus [see the snotty periphery], Pseudomonas [yellowish]--both very fast growing; The pinkish shit could be any number of things.
Judging from the flow path of that mess, it looks to me like there was a bacterial innoculation at the center, and the innoculant ran across the diameter of the petri.
A couple very important petri tips, IMO. . .
If you allow the agar to cool down to ~60 C or so before pouring [via warm oven, water bath, etc] the amount of resulting condensation will be reduced.
Pouring the petris in a stack ~10 or so high will also cut down on the condensation.
After the plates have cooled, flip the stack upside down. This is the best way to keep/store/incubate cultures, IME. As mycelium grows, additional condensation will occur; droplets will not run across the surface of the agar if they're in this orientation. An additional step that may be too much of a risk, depending on equipment/cleanliness, is to wipe any residual condensation from the bottom plate [the non-agar'd piece] during innoculation.
Condensation is something to be minimized. Even with clean innoculum, wet petris can wick things inside.


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Offlinellamaboy
the weasel thatsnagged the bee

Registered: 11/08/03
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Loc: Portland PNW
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Re: Question about agar contam *pic* [Re: Suntzu]
    #2216780 - 01/03/04 03:56 PM (12 years, 11 months ago)

because every singe petri dish ended up with this little bacterial concoction, do you think it was something that was in the sryinge, or something from the outside...the bacteria seems to have only been where there was water from the innoculation...or is it just that bacteria need standing water to grow, and thus, they grew on the innoculant water.


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OfflineSuntzu
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Re: Question about agar contam *pic* [Re: llamaboy]
    #2216885 - 01/03/04 05:22 PM (12 years, 11 months ago)

It really looks like it was in the innoculum.


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Offlinellamaboy
the weasel thatsnagged the bee

Registered: 11/08/03
Posts: 563
Loc: Portland PNW
Last seen: 7 years, 1 month
Re: Question about agar contam *pic* [Re: Suntzu]
    #2216957 - 01/03/04 06:17 PM (12 years, 11 months ago)

hrmmmmmm....that's what i was afraid of...do you think that it would be best to replace the syringe? if i used a different method, do you think that the bacteria would still get the foothold like that, or could the mycelia take over?


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OfflineSeussA
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Re: Question about agar contam *pic* [Re: llamaboy]
    #2220524 - 01/05/04 02:32 PM (12 years, 11 months ago)

> It really looks like it was in the innoculum.

My thoughts as well... also appears to be bacterial from the picture. Try using an antibiotic in your agar next time.


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