SEA-PHAGES | Top Agar contamination

Link to this post \| posted 23 Feb, 2024 21:50
jdaft@leeuniversity.edu	Has anyone ever had an issue with thermophile contamination in their top agar? We've cultured the thermophile to a PYCa plate and growth only occurs at 60C and not 37C. We think we narrowed it down to our calcium chloride and/or dextrose. Both were remade and filtered and we still are having contamination issues. Our host bacteria is M. foliorum, so our thermophile doesn't appear to be interfering with it's growth, but we still don't like have another bacteria in there with our host culture. ~Joe

Link to this post | posted 23 Feb, 2024 21:50

Has anyone ever had an issue with thermophile contamination in their top agar? We've cultured the thermophile to a PYCa plate and growth only occurs at 60C and not 37C. We think we narrowed it down to our calcium chloride and/or dextrose. Both were remade and filtered and we still are having contamination issues. Our host bacteria is M. foliorum, so our thermophile doesn't appear to be interfering with it's growth, but we still don't like have another bacteria in there with our host culture.
~Joe

Link to this post \| posted 24 Feb, 2024 15:26
viknesh	jdaft@leeuniversity.edu Has anyone ever had an issue with thermophile contamination in their top agar? We've cultured the thermophile to a PYCa plate and growth only occurs at 60C and not 37C. We think we narrowed it down to our calcium chloride and/or dextrose. Both were remade and filtered and we still are having contamination issues. Our host bacteria is M. foliorum, so our thermophile doesn't appear to be interfering with it's growth, but we still don't like have another bacteria in there with our host culture. ~Joe Can't say I've heard from the community about an issue like this. Can you share more about how you discovered it, since it doesnt grow at 37? Photos would be great too. Vic

Link to this post | posted 24 Feb, 2024 15:26

viknesh

jdaft@leeuniversity.edu
Has anyone ever had an issue with thermophile contamination in their top agar? We've cultured the thermophile to a PYCa plate and growth only occurs at 60C and not 37C. We think we narrowed it down to our calcium chloride and/or dextrose. Both were remade and filtered and we still are having contamination issues. Our host bacteria is M. foliorum, so our thermophile doesn't appear to be interfering with it's growth, but we still don't like have another bacteria in there with our host culture.
~Joe

Can't say I've heard from the community about an issue like this. Can you share more about how you discovered it, since it doesnt grow at 37? Photos would be great too.

Vic

Link to this post \| posted 11 Mar, 2024 13:46
jdaft@leeuniversity.edu	PYCa plate inoculated with contaminated top agar and incubated at 60C. 1.42Mb

Link to this post \| posted 11 Mar, 2024 13:46
jdaft@leeuniversity.edu	PYCa plate inoculated with contaminated top agar and incubated at 28C. 1.28Mb

Link to this post \| posted 11 Mar, 2024 13:49
jdaft@leeuniversity.edu	We did a Gram stain and it appears that the organism is Gram (-). We're not dismissing that it could be archaea, but we haven't looked into what it would take to confirm that.

Link to this post \| posted 18 Mar, 2024 14:53
viknesh	Thanks for sharing, Joe. I think this is going to be ver helpful for other faculty if they are seeing all their top agar get contaminated on a regular basis. Hvae you been able to narrow it down to your dextrose or calcium chrloride solution. Any more info you can share will be helpful for the community.

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Top Agar contamination