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A series of tubes. The tube with 2 micrograms/ml has growth, as does the tube with 4 micrograms/ml. The tubes with 8, 16, and 32 do not have growth
In a dilution test, the lowest dilution that inhibits turbidity (cloudiness) is the MIC. In this example, the MIC is 8 μg/mL. Broth from samples without turbidity can be inoculated onto plates lacking the antimicrobial drug. The lowest dilution that kills ≥99.9% of the starting inoculum is observed on the plates is the MBC. (credit: modification of work by Suzanne Wakim)
A 64 well plate; 8 rows and 12 columns. The concentration increases from left to fight. Each row has a different antibiotic. The MIC is determined by the lowest concentration with no growth as seen by a clear rather than dark look to the well. For clindamycin the MIC is above the highest concentration of 32 micrograms per mL. For Peniciliin the MIC is 0.06 and for Erythromycin it’s 8 micrograms per mL. The bottom row shows positive and negative controls.
A microdilution tray can also be used to determine MICs of multiple antimicrobial drugs in a single assay. In this example, the drug concentrations increase from left to right and the rows with clindamycin, penicillin, and erythromycin have been indicated to the left of the plate. For penicillin and erythromycin, the lowest concentrations that inhibited visible growth are indicated by red circles and were 0.06 μg/mL for penicillin and 8 μg/mL for erythromycin. For clindamycin, visible bacterial growth was observed at every concentration up to 32 μg/mL and the MIC is interpreted as>32 μg/mL. (credit: modification of work by Centers for Disease Control and Prevention)
A strip with numbers on a lawn of bacteria. The top of the strip has the highest concentration; the bottom has the lowest. Bacteria are able to grow at anything below 1.5 micrograms per mL
The Etest can be used to determine the MIC of an antibiotic. In this Etest, vancomycin is shown to have a MIC of 1.5 μg/mL against Staphylococcus aureus .
  • Compare and contrast MIC and MBC.

Resolution

Marisa’s UTI was likely caused by the catheterizations she had in Vietnam. Most bacteria that cause UTIs are members of the normal gut microbiota, but they can cause infections when introduced to the urinary tract, as might have occurred when the catheter was inserted. Alternatively, if the catheter itself was not sterile, bacteria on its surface could have been introduced into Marisa’s body. The antimicrobial therapy Marisa received in Cambodia may also have been a complicating factor because it may have selected for antimicrobial-resistant strains already present in her body. These bacteria would have already contained genes for antimicrobial resistance, either acquired by spontaneous mutation or through horizontal gene transfer, and, therefore, had the best evolutionary advantage for adaptation and growth in the presence of the antimicrobial therapy. As a result, one of these resistant strains may have been subsequently introduced into her urinary tract.

Laboratory testing at the CDC confirmed that the strain of Klebsiella pneumoniae from Marisa’s urine sample was positive for the presence of NDM, a very active carbapenemase that is beginning to emerge as a new problem in antimicrobial resistance. While NDM-positive strains are resistant to a wide range of antimicrobials, they have shown susceptibility to tigecycline (structurally related to tetracycline ) and the polymyxins B and E ( colistin ).

To prevent her infection from spreading, Marisa was isolated from the other patients in a separate room. All hospital staff interacting with her were advised to follow strict protocols to prevent surface and equipment contamination. This would include especially stringent hand hygiene practices and careful disinfection of all items coming into contact with her.

Marisa’s infection finally responded to tigecycline and eventually cleared. She was discharged a few weeks after admission, and a follow-up stool sample showed her stool to be free of NDM-containing K. pneumoniae , meaning that she was no longer harboring the highly resistant bacterium.

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Key concepts and summary

  • The Kirby-Bauer disk diffusion test helps determine the susceptibility of a microorganism to various antimicrobial drugs. However, the zones of inhibition measured must be correlated to known standards to determine susceptibility and resistance, and do not provide information on bactericidal versus bacteriostatic activity, or allow for direct comparison of drug potencies.
  • Antibiograms are useful for monitoring local trends in antimicrobial resistance/susceptibility and for directing appropriate selection of empiric antibacterial therapy.
  • There are several laboratory methods available for determining the minimum inhibitory concentration (MIC) of an antimicrobial drug against a specific microbe. The minimal bactericidal concentration (MBC) can also be determined, typically as a follow-up experiment to MIC determination using the tube dilution method.

Fill in the blank

The method that can determine the MICs of multiple antimicrobial drugs against a microbial strain using a single agar plate is called the ________.

Etest

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True/false

If drug A produces a larger zone of inhibition than drug B on the Kirby-Bauer disk diffusion test, drug A should always be prescribed.

false

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Short answer

How is the information from a Kirby-Bauer disk diffusion test used for the recommendation of the clinical use of an antimicrobial drug?

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What is the difference between MIC and MBC?

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Source:  OpenStax, Microbiology. OpenStax CNX. Nov 01, 2016 Download for free at http://cnx.org/content/col12087/1.4
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