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OfflineBaern
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Registered: 02/09/11
Posts: 20
Last seen: 3 years, 5 months
Agar Contamination or Weird Mycellial Growth? Pics
    #22275185 - 09/22/15 05:05 PM (8 years, 4 months ago)

Hey folks,

Getting back into mycology. Trying agar for the second time after a hiatus of a couple years. I built a flowhood, but I'm fairly sure I still got a lot of contams with my recent agar dishes. Before I confirm that analysis, here are some pics.








What is that strange, nearly translucent, perimeter around my inoculations, and what could account for it? Initially I thought it might be a foreward outpost of mycelium but it appears to be much too large for that now. I'm assuming contamination, but I can't tell of what kind. If anyone can shed some light on this perhaps I can do more to limit my contamination vectors in the future.


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Offlinevagblast
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Registered: 10/04/10
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Re: Agar Contamination or Weird Mycellial Growth? Pics [Re: Baern]
    #22275196 - 09/22/15 05:07 PM (8 years, 4 months ago)

Pics? Don't.see em


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OfflineBaern
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Re: Agar Contamination or Weird Mycellial Growth? Pics [Re: vagblast]
    #22275212 - 09/22/15 05:10 PM (8 years, 4 months ago)

Sorry, fixed.


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OfflineBaern
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Re: Agar Contamination or Weird Mycellial Growth? Pics [Re: Baern]
    #22275229 - 09/22/15 05:13 PM (8 years, 4 months ago)

Some more, not sure if holding them up helped them be visible.







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Offlinevagblast
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Re: Agar Contamination or Weird Mycellial Growth? Pics [Re: Baern]
    #22275308 - 09/22/15 05:28 PM (8 years, 4 months ago)

Yeah I'm pretty sure that's bacteria. Lots of it. Where did u take the tissue from for those clones?


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OfflineBaern
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Re: Agar Contamination or Weird Mycellial Growth? Pics [Re: vagblast]
    #22275337 - 09/22/15 05:32 PM (8 years, 4 months ago)

Yeah I did a bit more digging myself and I believe you're 100% correct. I didn't wash my specimen before I did my transfer. Upon reflection, this seems rather elementary. Is there a specific type of solution I should be using to wash my specimens before transfer? I scalpel'ed from the base of a fruitbody stem.

**EDIT** The fruitbody was from an indoor culture. It's possible it could have picked up some bacteria from its surroundings before making it the clean room? I'm troubled that I could have botched my sterile technique so badly.


Edited by Baern (09/22/15 05:40 PM)


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Invisiblejbaby007
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I'm a teapot


Registered: 02/28/15
Posts: 1,026
Re: Agar Contamination or Weird Mycellial Growth? Pics [Re: Baern]
    #22275557 - 09/22/15 06:19 PM (8 years, 4 months ago)

Check out this cloning tek. Frank recommends using a SAB rather than a flowhood since it can potentially blow around contams. There's no need to wash the mushroom you want to clone because it was fruited in open air, which isn't sterile to begin with.

Just make sure you are tearing the fruit body open rather than cutting into it, that way you don't drive contams into the mushroom and make sure to follow proper sterile techniques. You'll more than likely need to make a few transfers before obtaining a clean culture to work with.


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OfflineBaern
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Re: Agar Contamination or Weird Mycellial Growth? Pics [Re: jbaby007]
    #22278196 - 09/23/15 10:43 AM (8 years, 4 months ago)

Thanks for the link to that tek. I believe my fatal error occurred when I was using my scalpel. Thinking back, what was covered in the tek about tearing and scraping was clearly stated in RR's vids. I totally cut a wedge out of the surface of my fruitbody. I feel kinda dumb because I was thinking as I was doing it "Gee this seems like it would be a bit prone to contams.... w/e". Time for round two. I was really perplexed, because otherwise I think my sterile technique was pretty solid.


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Offlinevagblast
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Re: Agar Contamination or Weird Mycellial Growth? Pics [Re: Baern]
    #22278316 - 09/23/15 11:13 AM (8 years, 4 months ago)

When I clone I just break the stem in half and scrape a peace off the middle. Works great for me


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OfflineBaern
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Re: Agar Contamination or Weird Mycellial Growth? Pics [Re: vagblast]
    #22282512 - 09/24/15 08:11 AM (8 years, 4 months ago)

Thanks guys! I got another harvest today. I'm gonna clone again and see if I can't do a better job this time, heh.


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OfflinePeloefuego
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Registered: 06/24/17
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Re: Agar Contamination or Weird Mycellial Growth? Pics [Re: jbaby007]
    #24482627 - 07/14/17 11:54 AM (6 years, 6 months ago)

I transferred small pieces of a healthy mycelium I had stored for nearly 4 moths in the fridge into new and sterile agar plates but got a very similar contamination to the ones in Baern pictures.

Can you get contams from a mycelium on agar plate that was on the fridge for 4 months?
I had my agar plate wrapped in plastic film inside the fridge.


Edited by Peloefuego (07/14/17 11:56 AM)


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