First of all remember that biofilms are heterogenous. Therefore, to test these against any antimicrobials will be next to impossible as there will be several "Shielding " parameters which you would have to account for in order to validate your results. Some of these parameters you may not even know and hence you can not satisfactorily draw any inference from your observations.
If the biofilm is a monospecies one, you can use our method named BioTimer assay developed to evaluate biofilm population without sample manipulation.
you can see the following papers:
1-Influence of sub-inhibitory antibiotics and flow condition on Staphylococcus aureus ATCC 6538 biofilm development and biofilm growth rate: BioTimer assay as a study model.
Berlutti F, Frioni A, Natalizi T, Pantanella F, Valenti P.
J Antibiot (Tokyo). 2014 Nov;67(11):763-9. doi: 10.1038/ja.2014.66. Epub 2014 May 28.
2-BioTimer Assay, a new method for counting Staphylococcus spp. in biofilm without sample manipulation applied to evaluate antibiotic susceptibility of biofilm.
Pantanella F, Valenti P, Frioni A, Natalizi T, Coltella L, Berlutti F.
J Microbiol Methods. 2008 Dec;75(3):478-84. doi: 10.1016/j.mimet.2008.07.027. Epub 2008 Jul 30.
Thank you all.
I found this new article on the internet where it described the use of sugar coating to increase the metabolism of the microbes within the biofilm. This makes antibiotics more susceptible. Does anyone have a method or have more knowledge about this?
Hi. Another MBEC method would be to culture biofilms in wells of titre plates, perform serial dilution with antimicrobials (one at a time) and growth medium (as done with MBC determinations), followed by incubation, dislodging of biofilms with any suitable method (to obtain life cells - if any), removal of medium with pipette and plating. Surviving cells from biofilms post-drug exposure would grow colonies or absence of colonies would help determine your concentrations (effective and antibiofilm concentrations). If not plating, skip the dislodging part and proceed with removal of medium and staining protocols to help in quantification of life cells, dead cells or/and matrix to obtain your required concentrations. Of course optimization and validation is required for each step. Please consider if your antimicrobial is a known drug with antibiofilm property or otherwise, as its ability to penetrate the matrix plays a role in combating biofilms too. Increasing the MO's metabolism and thus sensitivity may only work if the antimicrobial is able to penetrate the matrix and reach the MO within it in the first place and also that its MOA is targeting the metabolic pathways. Good luck.
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