Hello everyone, I am having difficulty growing Listeria monocytogenes biofilms using the Calgary Biofilm Device (CBD) for the purpose of completing a MBEC assay. It seems like the biofilms are not adhering to the pegs, as tested by a CV staining assay. I have tried using different broths (brain heart infused (BHI) broth, BHI broth with 0.1% glucose, BHI broth with 1% glucose, BHI broth with 2% glucose, Tryptic soy broth (TSB) and TSB with 0.6% yeast extract), different incubation temperatures (37 degrees celsius and 30 degrees celsius), different rotation speeds (110 rpm, 200 rpm and stationary) as well as different incubation times (24hrs, 48 hrs, & 72hrs (changing media every 24hrs)). Iv'e tried all of these different methods with no success. Does anyone have any suggestions or different methodology? Thank you!
Dear Joelle, in the following paper, NaCl addition and at low temp (+4 C), the biofilm formation of ATCC19112 increased. However, we are working with biofilm formation abilities of S. aureus and S. epidermidis strains and the biofilm formers of these strains OD570 values are higher than 0.5. In that paper, OD570 values was very low.
Braz J Microbiol. 2013 Mar 19;44(1):51-5. doi: 10.1590/S1517-83822013005000004. eCollection 2013.
Formation of biofilm by Listeria monocytogenes ATCC 19112 at different incubation temperatures and concentrations of sodium chloride.
Are you sure this strain really strong biofilm former?
Best regards.
The following chapter also describes biofilm formation of Listeria monocytogenes.
Control of Listeria monocytogenes in the Processing Environment by Understanding Biofilm Formation and Resistance to Sanitizers, Chapter 21, pp 251-261
by Stavros G. Manios and Panagiotis N. Skandamis
in Kieran Jordan et al. (eds.), Listeria monocytogenes: Methods and Protocols, Methods in Molecular Biology, vol. 1157, 2014
Thank you Mert Sudagidan for your valuable input. You are right, this strain of Listeria monocytogenes is probably not a strong biofilm former. We will try with another strain for these experiments, thanks again!
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