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Offlineswinghigh
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agar plate questions.
    #26882484 - 08/15/20 10:58 PM (3 years, 5 months ago)

ive got a few plates i need some input on..... these plates are 5 days in and on there 3rd transfer from a liquid live culture (Brazilian). onto MEA the plates may seem a bit darker on one side because the agar is a tad bit deeper on that side do to an unbalanced work table










someone had suggested there may be a bacterial contamination on the plates, i cant seem to see it but he did point out a problem area that he said looked like a halo area.... i dont disagree with this persons statement but i was looking to get a bit more confirmation on it....... he states that on the plates show below..."I DO see a halo: just behind the leading edge in the nonuniform section of the circle... there are 2 very small tips just past the haze: its there...  try grabbing just the tip... you can see the line continues around to the left underneath the culture... liquid cultures are especially susceptible to bacteria"









i do see where the person has stated they see a sign of contamination but i cant justify if it actually is a contamination or just lighting/picture isnt good.....ive put my transfer off untill tomorrow to try and get a better understanding of the issue that was brought up and how to keep it away from my next transfer, as i thought these plates where going to be great and looked amazing to my knowledge.


Edited by swinghigh (08/15/20 11:11 PM)


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OfflineTrust_d Cultivator
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Re: agar plate questions. [Re: swinghigh]
    #26882494 - 08/15/20 11:08 PM (3 years, 5 months ago)

I see no obvious contam.  Nice cultures ya got there :cool:


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InvisibleGoatrider
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Re: agar plate questions. [Re: Trust_d Cultivator]
    #26882497 - 08/15/20 11:15 PM (3 years, 5 months ago)

That`s the case.
You don`t see it.

That`s the hardest thing with agar works,
to recognize starting contams, not related to
any changes of colour or something.
I have hard times giving advice on pictures.
There could be something, yes.

But really hard to tell for me.


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Offlineswinghigh
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Re: agar plate questions. [Re: Trust_d Cultivator]
    #26882498 - 08/15/20 11:17 PM (3 years, 5 months ago)

i have done two tester totes from one of those plates i transferred to mother jars of oats then to coir and these totes where put into colonization on the 9th of this month and one has colonized and ones a few days behind but thats because the tote is bigger.







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Offlineswinghigh
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Re: agar plate questions. [Re: Goatrider]
    #26882507 - 08/15/20 11:21 PM (3 years, 5 months ago)

couldnt agree more its not easy from pictures and im a very indecisive person so im always second guessing or worrying about things lol. but from knowledge on agar i do see the area this person was concerned with but i just cant seem to distinctively tell if its actually a contamination issue lol sheeeet.


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Offlinedirtybirdx
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Re: agar plate questions. [Re: swinghigh]
    #26882510 - 08/15/20 11:22 PM (3 years, 5 months ago)

Definitely some wispyness near that uncolonized area.  A contaminate could be a reason for mycelium not growing there.

Aside from that, why have you grown the culture out so much?  If you intend to do 4 transfers, as the typical standard for due diligence requires, then the idea is to take a transfer as soon as you spot your healthy mycelium.  The idea is to ‘run away’ from the contaminates to new plates before they are able to get hold.

I never let any culture grow to the edge of the plate, to reduce risk of contam, but if I was satisfied that my 3rd transfer was clean and safe to use as inoculate, I would let it grow until it’s a bit from the edge of the plate (or agar), and then stall it in the fridge until you’re ready to use it.


--------------------
Need help?




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Offlineswinghigh
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Re: agar plate questions. [Re: dirtybirdx]
    #26882512 - 08/15/20 11:27 PM (3 years, 5 months ago)

ya that tote that hasnt fully colonized is bigger so its not done colonizing it needs a few more days the other tote finished so fast becasuse its much smaller substrate level
these where stored away in the fridge for awhile, they where going to be used as transfer to jars but i stored them away. just took them out tonight to start a transfer when someone suggested they are bacterial.


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OfflineShroom Mafia
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Re: agar plate questions. [Re: swinghigh]
    #26882527 - 08/15/20 11:47 PM (3 years, 5 months ago)

That's some healthy looking mycelium!

Doesn't look bacterial to be honest but it's hard to tell from this perspective. Is there any chance that it could be related to the sector of mycelium itself? Like others have mentioned, it looks kind of wispy in that region, and I can relate to what you describe as the "halo area" you describe, as I've seen sometimes on my own plates almost like an area around the mycelium for future growth. Almost looks sometimes like it's trying to lay a foundation, but maybe someone else who is more knowledgeable could elaborate as to if that's even a possible scenario.

What is the agar recipe, if I may ask? Also, did the liquid culture have any additives to it, or just straight water and mycelium?


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Offlineswinghigh
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Re: agar plate questions. [Re: Shroom Mafia]
    #26882532 - 08/15/20 11:57 PM (3 years, 5 months ago)

its a pre mix i buy from a local mushroom farm  35.7% Malt Extract 1.8% Pancreatic Digest of Gelatin (Peptone G) 35.7% Dextrose 26.8% Agar Agar

and as far as i know the liquid culture is  only water and Mycelium. its from a very reputable source known for good clean cultures.

either way im going to take some sectors tomorrow from those plates around the edges, probably do 4 new plates........ if anyone sees a nice spot that catches there eyes lemme know


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