Minimum bactericidal concentration

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The minimum bactericidal concentration (MBC) is the lowest concentration of an antibacterial agent required to kill a particular bacterium. [1] It can be determined from broth dilution minimum inhibitory concentration (MIC) tests by subculturing to agar plates that do not contain the test agent. The MBC is identified by determining the lowest concentration of antibacterial agent that reduces the viability of the initial bacterial inoculum by ≥99.9%. [2] The MBC is complementary to the MIC; whereas the MIC test demonstrates the lowest level of antimicrobial agent that inhibits growth, the MBC demonstrates the lowest level of antimicrobial agent that results in microbial death. This means that even if a particular MIC shows inhibition, plating the bacteria onto agar might still result in organism proliferation because the antimicrobial did not cause death. Antibacterial agents are usually regarded as bactericidal if the MBC is no more than four times the MIC. [3] [4] Because the MBC test uses colony-forming units as a proxy measure of bacterial viability, it can be confounded by antibacterial agents which cause aggregation of bacterial cells. Examples of antibacterial agents which do this include flavonoids [4] and peptides. [5] [6]

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Virtual colony count (VCC) is a kinetic, 96-well microbiological assay originally developed to measure the activity of defensins. It has since been applied to other antimicrobial peptides including LL-37. It utilizes a method of enumerating bacteria called quantitative growth kinetics, which compares the time taken for a bacterial batch culture to reach a threshold optical density with that of a series of calibration curves. The name VCC has also been used to describe the application of quantitative growth kinetics to enumerate bacteria in cell culture infection models. Antimicrobial susceptibility testing (AST) can be done on 96-well plates by diluting the antimicrobial agent at varying concentrations in broth inoculated with bacteria and measuring the minimum inhibitory concentration that results in no growth. However, these methods cannot be used to study some membrane-active antimicrobial peptides, which are inhibited by the broth itself. The virtual colony count procedure takes advantage of this fact by first exposing bacterial cells to the active antimicrobial agent in a low-salt buffer for two hours, then simultaneously inhibiting antimicrobial activity and inducing exponential growth by adding broth. The growth kinetics of surviving cells can then be monitored using a temperature-controlled plate reader. The time taken for each growth curve to reach a threshold change in optical density is then converted into virtual survival values, which serve as a measure of antimicrobial activity.

Agar dilution is one of two methods used by researchers to determine the Minimum Inhibitory Concentration (MIC) of antibiotics. It is the dilution method most frequently used to test the effectiveness of new antibiotics when a few antibiotics are tested against a large panel of different bacteria.

<span class="mw-page-title-main">Murepavadin</span> Chemical compound

Murepavadin also known as POL7080 is a Pseudomonas specific peptidomimetic antibiotic. It is a synthetic cyclic beta hairpin peptidomimetic based on the cationic antimicrobial peptide protegrin I (PG-1) and the first example of an outer membrane protein-targeting antibiotic class with a novel, nonlytic mechanism of action, highly active and selective against the protein transporter LptD of Pseudomonas aeruginosa. In preclinical studies the compound was highly active on a broad panel of clinical isolates including multi-drug resistant Pseudomonas bacteria with outstanding in vivo efficacy in sepsis, lung, and thigh infection models. Intravenous murepavadin is in development for the treatment of bacterial hospital-acquired pneumonia and bacterial ventilator-associated pneumonia due to Pseudomonas aeruginosa.

References

  1. Amyes S et al. Antimicrobial Chemotherapy: Pocketbook. CRC Press, 1996 ISBN   9781853173899 Page 25
  2. National Committee for Clinical Laboratory Standards (1999). Methods for determining bactericidal activity of antimicrobial agents : approved guideline M26-A (PDF). Vol. 19. Arthur L. Barry et. al. Wayne, PA: National Committee for Clinical Laboratory Standards. ISBN   1-56238-384-1. OCLC   1124514908.
  3. French GL (2006). "Bactericidal agents in the treatment of MRSA infections--the potential role of daptomycin". Journal of Antimicrobial Chemotherapy. 58 (6): 1107–17. doi:10.1093/jac/dkl393. PMID   17040922.
  4. 1 2 Cushnie TP, Cushnie B, Echeverría J, Fowsantear W, Thammawat S, Dodgson JL, Law S, Clow SM (2020). "Bioprospecting for antibacterial drugs: a multidisciplinary perspective on natural product source material, bioassay selection and avoidable pitfalls". Pharmaceutical Research. 37 (7): Article 125. doi:10.1007/s11095-020-02849-1. PMID   32529587. S2CID   219590658.
  5. Suarez M, Haenni M, Canarelli S, Fisch F, Chodanowski P, Servis C, Michielin O, Freitag R, Moreillon P, Mermod N (2005). "Structure-function characterization and optimization of a plant-derived antibacterial peptide". Antimicrobial Agents and Chemotherapy. 49 (9): 3847–3857. doi:10.1128/AAC.49.9.3847-3857.2005. PMC   1195432 . PMID   16127062.
  6. Robert É, Lefèvre T, Fillion M, Martial B, Dionne J, Auger M (2015). "Mimicking and understanding the agglutination effect of the antimicrobial peptide thanatin using model phospholipid vesicles". Biochemistry. 54 (25): 3932–41. doi:10.1021/acs.biochem.5b00442. PMID   26057537.