Dear colleagues,
I am following the MBEC Protocol from Harrison et al (2010) to test the effect of an Antimicrobial Peptide (LyeTx I) against Oral Biofilm. In my experimental design I want to compare the biofilm and planktonic cell susceptibility, therefore, when looking at the core protocol, Harrison et al suggest that "biofilms and planktonic cells should be treated with a "Neutralizing Agent" in order to reduce the toxicity of it.
My question is: When using the MBEC Protocol to test Antimicrobial Peptides (AMPs) against Oral Biofilms, should I use a neutralizing agent?
References I am using for my experimental design:
- Harrison JJ, Stremick CA, Turner RJ, Allan ND, Olson ME, Ceri H. Microtiter susceptibility testing of microbes growing on pegs lids: a miniaturized biofilm model for high-throughput screening. Nature Protocols. 2010; Vol. 5, No. 7.
- Harrison JJ, Turner RJ, Ceri H. High-throughput metal susceptibility testing of microbial biofilms. BMC Microbiology. 2005; Jan 5(1): 53
Thank you very much, and best regards!
Hi Edison,
the use of neutralizing agent is used to reduce the toxicity of your antimicrobial compound during the transfer from challenge plate to recovery plate. It is particularly important if you are evaluating the effect of your antimicrobial peptide with a precise timing.
As neutralizing agent, you should use a compound that neutralize the toxicity of your antimicrobial peptide, without affecting the viability of the bacterial cells.
This is the composition of the Universal Neutralizer used for biocide testing:
1.0 g L-Histidine
1.0 g L-Cysteine
2.0 g Reduced glutathione
Hope it helps,
Emanuele
Dear Emanuele, your suggestions are very wellcome. Thank you very much for helping me on this topic.
Best regards!
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