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InvisibleTrippyTreez
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Registered: 11/23/19
Posts: 2
First shot at Agar, Obvious contams / Potential Tomentose growth?
    #26340466 - 11/23/19 02:49 PM (4 years, 2 months ago)

Little background. I'm a newbie, the microbiology background I have is in brewing compost teas and cannabis. This was my first go at Agar, and admittedly I was sloppy on my movements in my SAB (Shit was tough the first go around).

This was a MS Syringe into Agar.

I used a pre mixed MAE mix - I poured in a SAB. I did not have a inoculation loop for this run. I poured slightly too hot. I had condensation that I removed with a warm glass of water. Which came back from temp swing in the warmer room I brought them into. I did NOT have para-film at the time and used micro-pore tape. I did not have a face mask/surgical mask at the time.

Basically this was a newbie first run, learning about my agar, pressure cooking, sterility etc. You name it I had to figure it out, but I figured I'm not scared of trying something like Agar so I dove right in.

My most recent pours with all the correct equipment, proper agar mixtures, surgical masks, inoculation loop, towel on bottom of the SAB, a Riser in the SAB on the towel, etc. Turned out great (so far) we'll see in the next couple days but they look pro as fuck, and I was pretty damn proud of myself to bounce back from this first run (which looks like it is totally fucked).

My first question is.. Are the saucer like mycelium growth actually Tementose Mycelium?

If you look at the pictures I took under the microscope, you can see small little, what look like water drops on the very end. From my reading people have claimed that to be mold. I couldnt find enough microscopic mycelium pictures to verify if this was in fact a contamination or not (pin mold etc) Ive ruled out cobweb mold as its not grey, everything is extremely white.

I must add... when these germinated, the small little starting patch looked like a white spiked ball. It didnt seem like a rhizomorphic mycelium.

The other agar dish, seems to have this mold or Tomentose mycelium piggy backing a rhizomorphic mycelium. You can see the strands poking out just on the edges of it. I've included a microscope shot of the rhizhomorphic looking mycelium as well. It looks like the end of a find brissled brush/broom. If you look closely you can also see small, what appear to be, "spores" as well.

It's also quite obvious to me that I have a bacterial contamination in one if not both of the agar dishes. 

I'd appreciate any advice on the contamination's present in my dishes. I would like to learn from this first run so I know what to look out for. I've already adjusted my sterility tek, so the next batch hopefully will come out great.

Thanks!

-trippy



Edited by TrippyTreez (11/23/19 02:58 PM)


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InvisiblebodhisattaMDiscordReddit
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Re: First shot at Agar, Obvious contams / Potential Tomentose growth? [Re: TrippyTreez]
    #26340483 - 11/23/19 03:01 PM (4 years, 2 months ago)

Sometimes mycelium grows like that I'd transfer some of it and see how it continues to grow. Pictures 7&8 look good imo


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InvisibleTrippyTreez
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Registered: 11/23/19
Posts: 2
Re: First shot at Agar, Obvious contams / Potential Tomentose growth? [Re: bodhisatta]
    #26340494 - 11/23/19 03:05 PM (4 years, 2 months ago)

Quote:

bodhisatta said:
Sometimes mycelium grows like that I'd transfer some of it and see how it continues to grow. Pictures 7&8 look good imo




Thank you Bod. Appreciate you taking the time to look at it.

Pictures 7 & 8 are the very outlying edge of the picture 6's growth. Which is the most gnarly looking to me (what do I know)

I was thinking I might be able to pull a microscopic bit of it off that edge and transfer it. I'll transfer an outlying edge of the tomentose looking saucer growth as well, and see where that takes me.

I was honestly slightly concerned about health risks associated with the whiter/fluffier mold. I forgot where I read it but some people were claiming that kind of growth could be an opportunistic mold (forgot what the name was)

Regardless i'll transfer it in the SAB and see where it takes me.

Thanks again for the response, and thank you for all of the knowledge you've dropped on here.

-trippy


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