I am using RCM and ATCC 2107 media for culturing C. chauvoei but sevaral times I get mixed culture even while reviving from stock pure culture. for ATCC 2107 i am using anaerobic gas pack also. Suggest me the protocol for culturing this organism without any contamination.
Hi,
In the past I have had some issues with ATCC stocks being slightly contaminated, or even the media that comes along with them. Recently, I have had problems with contamination while growing Clostridium species in the same way as you- on RCM and in anaerobic jars with gas packs. What I did was prepare my own RCM, not what ATCC gave me, streaked my culture into 3-4 quadrants, and selected the predominate species on the plate. The previous freezer stocks were not grossly contaminated, so it was easy to find the predominate species. Simple to do, and I also made several pure stocks of my own.
I am sure you already know this, but you might also check the O-ring around the jar to make sure it still is tight. In addition, replace the gas packs as suggested on the packs you are using. I hope this helps
Try with nitrogen gas. Inject the inoculated vials with nitrogen and seal it and incubate.
Please check the components of anaerobic gas packs, they could be source of contamination some time when you do everything right.
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