ritualism example What is difference between minimum inhibitory concentration (MIC …

ritualism example What is difference between minimum inhibitory concentration (MIC …

Microbiology and Cell Biology, Medicinal Chemistry - Emery PharmaMicrobiology and Cell Biology, Medicinal Chemistry - Emery Pharma

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Minimum Inhibitory Concentration (MIC) and

Minimum Bactericidal Concentration (MBC) Assay

Minimum Inhibitory Concentration (MIC) assay

Minimum Inhibitory Concentration (MIC) assay tests for antimicrobial susceptibility and sensitivity by measuring inhibition of growth of bacteria or fungi as a function of concentration. Emery Pharma routinely provides antibiotic susceptibility testing services using disc diffusion, agar dilution and broth microdilution MIC methods according to CLSI protocols (Check out our MIC guide in the resources section). Test articles can be natural or synthetic, mixtures or purified. Hundreds of bacterial strains , including multi drug-resistant clinical isolates, ESKAPE pathogens, and multiple fungal strains , are available in our inventory for immediate testing.

Below is an illustration of a panel representing Broth Microdilution MIC for 8 antimicrobials tested against a susceptible bacterial strain:

Minimum Inhibitory Concentration (MIC), 96-Well Plate

Minimum Bactericidal/Fungicidal Concentration (MBC & MFC)

Minimum Bactericidal Concentration (MBC) or Minimal Fungicidal Concentration (MFC) is the lowest concentration of an antibacterial drug that results in a 99.9% reduction in the initial microbial density. EP routinely performs antimicrobial effectiveness testing services which normally includes MBC (minimum bactericidal concentration) / MFC (minimum fungicidal concentration) in conjunction with MIC (minimum inhibitory concentration) to distinguish if antimicrobial compounds are bactericidal or bacteriostatic.

Below is a schematic representation of MIC and MBC assays for a test article:

Minimum Bactericidal Concentration (MBC) Assay

  1. As shown, the test article is serially diluted in the presence of bacterial inoculum, and the MIC demonstrates the lowest concentration of the test article that inhibits the bacterial growth. At this dilution the test article is bacteriostatic.
  2. MBC is the lowest concentration of a test article required to kill 99.9% of an initial bacterial inoculum. At this dilution the test article is bactericidal.

Click on the image below to watch a video of our Biology scientists explaining the basics of the MIC assay!

Minimum Inhibitory Concentration (MIC) Video

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Minimum Bactericidal Concentration (MBC) Test


The MBC test determines the lowest concentration at which an antimicrobial agent will kill a particular microorganism.  The MBC is determined using a series of steps, undertaken after a  Minimum Inhibitory Concentration (MIC)  test has been completed.

MBC testing is useful for comparing the germ-killing activity of several antimicrobial agents at once.

 

 


Summary of the Minimum Bactericidal Concentration Test
  • A pure culture of a specified microorganism grown overnight, then diluted in growth-supporting broth (typically Mueller Hinton Broth) to a concentration between 1 x 10^5 and 1 x 10^6 cfu/ml.
  • A stock dilution of the antimicrobial test substance is made at approximately 100 times the expected  MIC  (if known).
  • Further 1:1 dilutions are made in test tubes or 96 well microtiter plates.
  • All dilutions of the test product(s) are inoculated with equal volumes of the specified microorganism.
  • A positive and negative control tube or well is included for every test microorganism to demonstrate adequate microbial growth over the course of the incubation period and media sterility, respectively.
  • An aliquot of the positive control is plated and used to establish a baseline concentration of the microorganism used.
  • The tubes or microtiter plates are then incubated at the appropriate temperature and duration.
  • Turbidity indicates growth of the microorganism and the MIC is the lowest concentration where no growth is visually observed.
  • To determine the MBC, the dilution representing the MIC and at least two of the more concentrated test product dilutions are plated and enumerated to determine viable CFU/ml.
  • The MBC is the lowest concentration that demonstrates a pre-determined reduction (such as 99.9%) in CFU/ml when compared to the MIC dilution.

Strengths of the Minimum Bactericidal Concentration Test
  • The MBC test allows determination of the minimum concentration of an agent necessary to achieve a bactericidal effect. It is worth noting, however, that the duration of time the antimicrobial is in contact with the test organism is quite long for this method, on the order of 18 hours. Thus, the test truly does determine the minimum concentration needed to kill the test organism, since all other parameters are conducive to biocidal effect. 
  • The MBC test can be a good and relatively inexpensive tool to rank a great number of antimicrobial agents by potency, for screening purposes.
  • The MBC test can be used to evaluate formulation problems wherein the formulator suspects that the active ingredient is being “bound up” by other ingredients. The theory is that the MBC will be worse for a formula that has a portion of its active ingredient chemically combined with other ingredients, thus not available to kill microorganisms in the suspension.
  • The test parameters for the MBC are easy to control in the laboratory, so comparisons can be made fairly easily between various antimicrobial agents tested under the same conditions and their respective effects on specific microorganisms.

Weaknesses of the Minimum Bactericidal Concentration Test
  • An MBC must be determined for each microorganism individually as the antimicrobial will likely have different MBC values for different test microorganisms.
  • MBC testing results will not determine the concentration of the antimicrobial needed to disinfect or sanitize microorganisms at a short contact time, such as 10 minutes.
  • More nutritive growth media such as Tryptic Soy Broth can negatively affect MBC values. Mueller Hinton Broth is the recommended broth for this method.

Microchem Laboratory routinely runs minimum bactericidal concentration (MBC) tests for its customers. The laboratory is well versed in the science of antimicrobial testing in general, and applies this knowledge to each and every study we undertake.

If you are interested in MBC testing, you are likely in the research and development rather than claim substantiation phase. Microchem Laboratory has an excellent track record of helping companies move from screening to GLP product testing.

 

Microchem Laboratory Is:

An EPA and FDA GLP-compliant testing organization staffed by skilled, experienced microbiologists and chemists.  Microchem can help speed your project to completion.

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