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Offlinecelticcelery
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Contam Found in SAB
    #26522335 - 03/07/20 02:36 PM (3 years, 10 months ago)

So after countless contaminations from transfers and spore inoculation, I decided to test my SAB for contaminants by doing an empty transfer.

By this I mean placing a sterilized petri dish in a SAB, then opening that petri dish and cutting that dish with a flame sterilized knife, then wrapping the dish with saran wrap.
After 2 weeks a familiar contamination resulted.

What contaminant is this and how should I deal with contamination in my SAB? Should I change up my procedure?

These are pictures of the contaminant when I performed an empty transfer,



While this is a picture of the contaminant when I did an agar transfer


Any advice and insight appreciated


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OfflineBooShow
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Re: Contam Found in SAB [Re: celticcelery]
    #26522347 - 03/07/20 02:48 PM (3 years, 10 months ago)

It looks like trich to me but it may be something else. Make sure you wipe down your SAB with iso or some type of disinfectant each time before you use it. Also, are there any drafts happening in the room you're working in? Drafts and air currents blow around unwanted contaminants and mold spores and will fuck you, even in a SAB. Do you flame sterilize in the SAB or outside of it? There could be tons of factors to why you're getting contams.


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You are what is. That's all.


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Offlinecelticcelery
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Re: Contam Found in SAB [Re: BooShow]
    #26522397 - 03/07/20 03:27 PM (3 years, 10 months ago)

So my SAB prep procedures goes as.

  • I wipe down only inside of SAB with 70% alcohol
  • I spray inside of SAB with a solution of (50% alcohol 50% H20)
  • Wait a few seconds
  • Put petri dishes in SAB
  • Flame sterilize outside of SAB before putting in
  • Note I do not wear gloves or mask when performing
  • Could it be the cloth?


This is a photo of my setup, it is in the basement.


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OfflineBooShow
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Re: Contam Found in SAB [Re: celticcelery]
    #26522419 - 03/07/20 03:41 PM (3 years, 10 months ago)

Quote:

celticcelery said:
So my SAB prep procedures goes as.

  • I wipe down only inside of SAB with 70% alcohol
  • I spray inside of SAB with a solution of (50% alcohol 50% H20)
  • Wait a few seconds
  • Put petri dishes in SAB
  • Flame sterilize outside of SAB before putting in
  • Note I do not wear gloves or mask when performing
  • Could it be the cloth?


This is a photo of my setup, it is in the basement.




Get rid of that towel, my friend. That could very well be your problem. Mold spores stick to cloth like shit on a blanket.


--------------------
You are what is. That's all.


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Offlinecelticcelery
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Re: Contam Found in SAB [Re: BooShow]
    #26522427 - 03/07/20 03:50 PM (3 years, 10 months ago)

Okay will try doing an empty inoculation without the towel and see if that resolves most of my issues.

Do you think that having a dirty basement could also contribute to contam? Like should I try to clean up that section of the area before doing any sab work?


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OfflineBooShow
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Re: Contam Found in SAB [Re: celticcelery]
    #26522434 - 03/07/20 03:59 PM (3 years, 10 months ago)

If you're working in an area that has little to no drafts, I believe you can inoculate in places that aren't the cleanest (using a SAB, of course). Work on a clean surface, make sure your SAB is clean, keep the drafts down, and keep your hands clean (I don't use gloves much either, just hand sanitizer. I do wear a face mask, though); oh, and if you're wearing long sleeves, roll them bitches up. All that said, it never hurts to have a nice, clean, and organized space to work in. It makes things easier.:thumbup::thumbup:


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You are what is. That's all.


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Offlinecelticcelery
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Re: Contam Found in SAB [Re: BooShow]
    #26522457 - 03/07/20 04:11 PM (3 years, 10 months ago)

alright, thanks BooShow


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OfflineBooShow
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Re: Contam Found in SAB [Re: celticcelery]
    #26522468 - 03/07/20 04:20 PM (3 years, 10 months ago)

No problem, man!  :super:  Let us know how it works out!


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You are what is. That's all.


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OfflineMrBovineJoni
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Re: Contam Found in SAB [Re: celticcelery]
    #26525160 - 03/09/20 08:43 AM (3 years, 10 months ago)

Quote:

celticcelery said:
So my SAB prep procedures goes as.

  • I wipe down only inside of SAB with 70% alcohol
  • I spray inside of SAB with a solution of (50% alcohol 50% H20)
  • Wait a few seconds
  • Put petri dishes in SAB
  • Flame sterilize outside of SAB before putting in
  • Note I do not wear gloves or mask when performing
  • Could it be the cloth?


This is a photo of my setup, it is in the basement.





a) Start using gloves and mask. Skin is covered with all kinds of stuff and microscopic pieces of skin drop constantly even if you wash frequently. One sneeze or even quick indirect breath will ruin the plate/work area because our mouths are full of all kinds of micro organisms.

b) Once you put everything in the SAB, leave it alone for 5-10 minutes for everything to settle ( I usually use this time to go to the other room and prepare - put on freshly washed long sleeved & tight fitting jumper/turtleneck, put on gloves and mask and wipe them with 70% iso). If not sure , I wipe with isopropyl some pieces/plates one more time while inside SAB before I transfer(more important for grain jars, as shit likes to live under the outer edge of the lid).

c) Nothing wrong with using towel if used properly but make sure it has been washed between each SAB session (unless you use it a lot(like every day), then can use it a few times). However, if you use one , I suggest buying a cooling rack and use it to elevate your working area above the towel (~2 inches I guess?).

4) Why do you wipe it with 70% and then spray it with 50%/50% solution? That's pointless. After wiping spray it with tap water and dish soap solution (a few drops per litre - it does not have to foam, just change the surface tension of the liquid) that slows down evaporation. That's what I do after reading quite a few SAB guides here, however, some TCs here just run SAB dry so it can work out easier if you have experience.


Edited by MrBovineJoni (03/09/20 09:00 AM)


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