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OfflineRo0k
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Assessment help, please. Contaminated Agar plates?
    #22392189 - 10/17/15 07:59 AM (8 years, 3 months ago)

I am wondering if I can tap the vast wealth of knowledge for an assessment of one of my plates.  It's a first transfer from a plate I made from a syringe.  Transferred on 10/10 from nutrient agar to nutrient agar.  To me it looks good, but I don't have the experience to tell whether it's healthy myc or not. 

As you can see in the first pic, there appears to be a healthy circular mycelium growth from the transfer, but there's also a secondary circle spreading out from the transfer that doesn't look as white and cloudy.  That's what I'm concerned about.  It looks stringy like myc, but it doesn't seem to have the vivid white of the stuff at the center.

Does it look like bacterial contamination?  Or am I just being thrown off by the difference in coloration?

Note: I think you can ignore the black flakes.  That's char from my flame sterilized knife that fell into the receiving plate.  Should be sterile as it was all burny. 

First image:


Under light for different contrast (I hope)



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OfflineGloogloos
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Re: Assessment help, please. Contaminated Agar plates? [Re: Ro0k]
    #22392353 - 10/17/15 09:09 AM (8 years, 3 months ago)

What species is it, I've had similar looking growth with Mexicana, but not with cubensis. 
Not in any of my dishes at least.

If it has any stringy or thread like growth it isn't bacteria, bacteria can come in many colors and shapes, but generally its a dense colored circle or, if there is fluid, its a soupy thick liquid.  If there are stringy things then you are dealing with some multi-cellular organism, more than likely a fungus.

It very could well be a another substrain from that multispore syringe(im assuming it was MS).  If you had pics of the other dish that you transferred from that would be awesome, and where did you transfer from in terms of the mycelium (from the very edge, just within the edge, towards the center, the actual center etc.)

It could also be a change in growth, mycelium will change in appearance as it grows outward, sometimes it becomes more strandy (rhizomorphic) and sometimes it becomes more fuzzy appearing(cottony?, I dont know the term lol).
But generally that happens in sectors, not around the whole thing.

My advice is as follows, if you don't have other dishes to work with and its all or nothing with this dish, isolate both myceliums into separate dishes.  If you do have other dishes and its not so important then observe, see what happens. The very worst that can happen is you learn something.

If it is indeed a contam you will find out pretty fast, and opening could lead to further contamination of your clean space, which is a risk that is seldom not worth taking.


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InvisiblebodhisattaMDiscordReddit
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Re: Assessment help, please. Contaminated Agar plates? [Re: Gloogloos]
    #22392370 - 10/17/15 09:14 AM (8 years, 3 months ago)



I would ignore the post above mine its like 95% wrong


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OfflineGloogloos
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Re: Assessment help, please. Contaminated Agar plates? [Re: bodhisatta]
    #22392491 - 10/17/15 09:53 AM (8 years, 3 months ago)

It's like 95% wrong? please clarify.  all you did was post pictures.


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Invisiblemicro
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Re: Assessment help, please. Contaminated Agar plates? [Re: Gloogloos]
    #22392525 - 10/17/15 10:01 AM (8 years, 3 months ago)

just tomentose vs. rhizomorphic growth i'd assume


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InvisiblebodhisattaMDiscordReddit
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Re: Assessment help, please. Contaminated Agar plates? [Re: Gloogloos]
    #22392537 - 10/17/15 10:04 AM (8 years, 3 months ago)

If bacteria is motile it will not form distinct colonies even if there's no extra moisture/water on the surface of your agar.

Some bacteria appear to form networks and are pesudo fungal like

As for substrains. They're just strains and if you grow ms you have 100-1000 strains easily from .25CC if you go right from a syringe with no dilutions.

Rhizomorphic and tomentose growth are much more a result of the nutrition % in agar rather than any sort of indication of performance.

Agar is there so we can open contaminated dishes to save cultures. If you know what you're doing you can open and use those plates. Don't try it with a flow hood though.

Also pictures are worth 1000 words


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OfflineRo0k
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Re: Assessment help, please. Contaminated Agar plates? [Re: bodhisatta]
    #22392613 - 10/17/15 10:15 AM (8 years, 3 months ago)

So is the first pic contaminated? The second one clearly is.


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Invisiblemicro
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Re: Assessment help, please. Contaminated Agar plates? [Re: bodhisatta]
    #22392616 - 10/17/15 10:16 AM (8 years, 3 months ago)

Quote:

bodhisatta said:
Some bacteria appear to form networks and are pesudo fungal likes




on TOP of the mycelium?

that looks like it's one thing that's growing, not fungi and a bacterial colony

that would make *two* circles >.>


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InvisibleMr.GuessWork
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Re: Assessment help, please. Contaminated Agar plates? [Re: bodhisatta]
    #22392759 - 10/17/15 10:49 AM (8 years, 3 months ago)

Quote:

bodhisatta said:
If bacteria is motile it will not form distinct colonies even if there's no extra moisture/water on the surface of your agar.







I didn't think you could see much motility on a solid medium, and I've definitely seen motile bacteria form colonies on solid agar. I thought you needed a semi-solid agar to see motility.


Quote:


Some bacteria appear to form networks and are pesudo fungal like





Are you talking about Streptomyces? The colonies look a bit different from mushroom mycelium. Here's a pic:



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OfflineRo0k
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Re: Assessment help, please. Contaminated Agar plates? [Re: Gloogloos]
    #22392834 - 10/17/15 11:06 AM (8 years, 3 months ago)

Strain B+

Took the sample from the leading edge from aa plate inoced from a ms syringe.

I had 2 plates of B+ and took 2 samples from each to make 4 plates. They all look roughly similar to this. No obvious secondary colonies that I can see, I'm mainly concerned with bacterial growth from under the transfer.


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Re: Assessment help, please. Contaminated Agar plates? [Re: Ro0k]
    #22392956 - 10/17/15 11:41 AM (8 years, 3 months ago)

Quote:

Ro0k said:
So is the first pic contaminated? The second one clearly is.



Both good


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InvisibleMad Season
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Re: Assessment help, please. Contaminated Agar plates? [Re: bodhisatta]
    #22392967 - 10/17/15 11:44 AM (8 years, 3 months ago)

:whathesaid: just look thin, and like an oversized leading edge lol.


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Re: Assessment help, please. Contaminated Agar plates? [Re: Mad Season]
    #22392978 - 10/17/15 11:48 AM (8 years, 3 months ago)


Bottom right was the same plate Cs5b and the other pic has Cs6b


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