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Offlinebradical
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Contam or healthy PE mycelium on agar after transfer?? * 1
    #27069074 - 12/02/20 10:12 PM (3 years, 1 month ago)

I did some transfer work on monday and went to check on my plates today to find some weird growth on one of my plates after transferring. The plate now has a ring around where the mycelium was and has a new small spot of growth on the outside of the plate. Does anyone one know if this is normal or is a contaminated? Should I toss the plate or try and make more transfers from the healthy mycelium? You can see the ring forming right beyond where i took transfers from. The small growth on the outside of the plate looks super fuzzy, somewhat uniform, and arial in growth. The ring that has formed around where i took the transfers is also very fuzzy and arial.



Edited by bradical (12/02/20 10:12 PM)


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OfflineSankhara
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Re: Contam or healthy PE mycelium on agar after transfer?? [Re: bradical]
    #27069254 - 12/03/20 03:01 AM (3 years, 1 month ago)

Unless you put something on the outside, which i doubt, thats a statelital contamination possibly due to improper sterile tek

Keep transfering


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Offlinepolaritymind
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Re: Contam or healthy PE mycelium on agar after transfer?? [Re: Sankhara]
    #27069388 - 12/03/20 07:04 AM (3 years, 1 month ago)

Interesting, could you elaborate what a satellital contamination is? I presume its bacteria but otherwise I know nothing about this, will be honest I have often succesfully grown with plates like that. Maybe its cause bacteria often arent as bad as mold and its not the full slimy plate contamination that stinks bacteria type either, which probably also helps.


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OfflineSankhara
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Re: Contam or healthy PE mycelium on agar after transfer?? [Re: polaritymind]
    #27069412 - 12/03/20 07:28 AM (3 years, 1 month ago)

They are contaminations that appear as a consequence of mostly improper sterile TEK.

They don't appear in the inoculation point, thats why they are statelital.

They could have been on your hands and fallen as a consequence of movements for example. Or they could get from the sides of the plates like in this case.

This is the reason it is not asvisable to use mycelium from the borders of the plates


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Offlinebradical
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Re: Contam or healthy PE mycelium on agar after transfer?? [Re: Sankhara]
    #27071053 - 12/04/20 01:20 AM (3 years, 1 month ago)

I was working in front of a flow hood and UV sterilized my work space before hand. Maybe a strand of hair touched the agar plate without me realizing it. Will the transfers I took from this plate be toast or do u think they have a chance of surviving?


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Offlinebradical
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Re: Contam or healthy PE mycelium on agar after transfer?? [Re: Sankhara]
    #27071055 - 12/04/20 01:24 AM (3 years, 1 month ago)

So from now on I should take transfers from more towards the center? I learned in my microbiology classes that its best to take transfers from the outside of the growth.


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OfflineMH5109
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Re: Contam or healthy PE mycelium on agar after transfer?? [Re: bradical]
    #27071059 - 12/04/20 01:32 AM (3 years, 1 month ago)

Your finger probably moved directly over the edge of the plate when you were taking the lid of or putting it on. It happens, and transfers are taken from the leading edge.


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Offlinebradical
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Re: Contam or healthy PE mycelium on agar after transfer?? [Re: MH5109]
    #27071101 - 12/04/20 02:20 AM (3 years, 1 month ago)

I made sure to wear gloves, sterilize with iso, and not touch anything besides the plate i was transferring from and the clean plate i was transferring to. How would me hovering over a plate in front of flow hood with clean hands produce this? I have a UVC sterilizing wand and placed it over this plate for about 20 min and have noticed no signs of the spot being destroyed, so does this mean that its maybe mycelium? It hasn't exponentially expanded growth in the past 24 hours so does this rule out the possibly of it being a Dactylium species growing?


Edited by bradical (12/04/20 02:32 AM)


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Offlinebradical
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Re: Contam or healthy PE mycelium on agar after transfer?? [Re: Sankhara]
    #27071111 - 12/04/20 02:28 AM (3 years, 1 month ago)

I left a high powered UVC light on for 45 min in my work area with these plates present before i did any transfer work so the outside of the plates where definitely sterile. I don't think its from me touching the outside of the plates and then hovering over the plate for too long considering i was wearing gloves, sterilized the gloves with alcohol before picking up said plates, and working in front of a flow hood.


Edited by bradical (12/04/20 02:37 AM)


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OfflineSankhara
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Re: Contam or healthy PE mycelium on agar after transfer?? [Re: bradical]
    #27071121 - 12/04/20 02:37 AM (3 years, 1 month ago)

UVC and alcohol sanitize, they do t sterilize. A spore could habe gotten there anyways.

Its nice that you have such equipment though


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OfflineSockadin
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Re: Contam or healthy PE mycelium on agar after transfer?? [Re: bradical]
    #27071122 - 12/04/20 02:37 AM (3 years, 1 month ago)

The UV wands are crap.

Looks like the mold halo around your original transfer is gonna be a bigger issue. Also a satellite contam isn't gonna be an issue because your probably not going to use that plate.

Did you transfer those wedges to new plates or knock up jars with them?


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Offlinebradical
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Re: Contam or healthy PE mycelium on agar after transfer?? [Re: Sockadin]
    #27071123 - 12/04/20 02:38 AM (3 years, 1 month ago)

I transferred those wedges to new plates.


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Offlinebradical
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Re: Contam or healthy PE mycelium on agar after transfer?? [Re: Sockadin]
    #27071126 - 12/04/20 02:41 AM (3 years, 1 month ago)

So the halo around my original growth is definitely mold?


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Offlinebradical
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Re: Contam or healthy PE mycelium on agar after transfer?? [Re: bradical]
    #27071130 - 12/04/20 02:45 AM (3 years, 1 month ago)

I really hope this isn't the case because i made like 20+ transfers from different plates after transferring from this plate. Gonna suck major balls if the mold spores being pushed onto me from the flow hood results in all my other plates being contamed. I guess i can just do lots of transfer work tho to get a clean culture going though.


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OfflineVoodoojuju
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Re: Contam or healthy PE mycelium on agar after transfer?? [Re: bradical]
    #27071149 - 12/04/20 03:11 AM (3 years, 1 month ago)

Quote:

bradical said:
So the halo around my original growth is definitely mold?




To add to this, what indicates that it is a mold halo and not mycelium? I'm curious as I sometimes get plates that do this and I'll end up with something that looks like a target.


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Offlinebradical
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Re: Contam or healthy PE mycelium on agar after transfer?? [Re: Voodoojuju]
    #27071156 - 12/04/20 03:18 AM (3 years, 1 month ago)

Phacts dude. If you look at my second picture you can clearly see the halo around the original growth. This new halo formed after i took transfer from the plate and looks super fuzzy and arial.


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Offlinepolaritymind
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Re: Contam or healthy PE mycelium on agar after transfer?? [Re: Sankhara]
    #27071313 - 12/04/20 07:04 AM (3 years, 1 month ago)

Quote:

Sankhara said:
They are contaminations that appear as a consequence of mostly improper sterile TEK.

They don't appear in the inoculation point, thats why they are statelital.

They could have been on your hands and fallen as a consequence of movements for example. Or they could get from the sides of the plates like in this case.

This is the reason it is not asvisable to use mycelium from the borders of the plates




Ok very interesting, I am learning new things.


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Offlinepolaritymind
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Re: Contam or healthy PE mycelium on agar after transfer?? [Re: polaritymind]
    #27071314 - 12/04/20 07:05 AM (3 years, 1 month ago)

And just to be clear about picture nr what are we talking about here with the mold halo?


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InvisibleLemgrub
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Re: Contam or healthy PE mycelium on agar after transfer?? [Re: polaritymind] * 1
    #27071535 - 12/04/20 10:15 AM (3 years, 1 month ago)

The small unconnected colony from the main cube sample. The growth is less uniform. Under a microscope you can see that cube hyphae grow in more straight lines and many contaminant fungi's hyphae look erratic in comparison. At least it seems that way to me. Top 2 pics are contaminant molds, bottom 2 are cubes. The one on the bottom left is cubensis next to a mold.




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OfflinePearl
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Re: Contam or healthy PE mycelium on agar after transfer?? [Re: Lemgrub]
    #27075129 - 12/06/20 11:41 AM (3 years, 1 month ago)

There's two conversations happening here...

One about satellite contamination that people keep rightly pointing out (the blob at 12 o'clock in pic 1).

And one about what the ring within the growth of the donor plate indicates (best seen in pic 2 - the ring matches up to the edge of the transfer wedges).

I'd wager the ring to be down to a change in storage of the plate post transfer.

If it was contamination it would be orders of magnitude unlikely for it to happen all around the leading edge of the myc and grow out so uniformly.

I'd be interested to know what the donor and transfer plates look like now...


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