1.

Why does the Kirby Bauer procedure require that the concentration of the bacteria be the same, the
stage of growth be constant, the growth of medium the same, and the concentration or amount of
drug in each disk constant?

- The Kirby-Bauer procedure is a standardized test that measures sensitivity of bacteria to

antibiotics by culturing bacteria on solid growth media surrounding sources of drug. The bacterial
concentration, medium growth, as well as the drug concentration in each disk needs to be
constant in order to get the most accurate results and measurements needed. Accuracy is a
crucial component in this test because clinicians use the test results from the procedure to
choose antibiotics effective against the specific bacteria causing a patient's infection. Using
specifically-targeted antibiotics helps decrease the frequency of drug-resistant bacteria evolving.

2. Why are E. coli, P. aeruginosa, and S. aureus used as standards in the Kirby Bauer method? Please
include your references.

- This is because Staphylococcus aureus and Escherichia coli are the most common pathogens
causing sepsis. Sepsis resulting from Pseudomonas aeruginosa infections may cause serious
complications among immunologically compromised patients.

Reference: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3819438/

3. What are the components of MacFarland standard? Give the exact contents of 10ml 0.1 to 1.0
MacFarland standard and its corresponding estimated bacterial cell count."

- A McFarland Standard is a chemical solution of barium chloride and sulfuric acid; the reaction
between these two chemicals results in the production of a fine precipitate, barium sulfate.

McFarland Standard 1% BaCl2 1% H2SO4 Approximate Bacterial Suspension /mL

0.1 0.01 9.91 7.6x107
0.5 0.05 9.95 1.5x108
1.0 0.10 9.90 3.0x108

10 mL
McFarland Standard 1% BaCl2 1% H2SO4 Approximate Bacterial Suspension
0.1 0.1 99.1 7.6x108
0.5 0.5 99.5 1.5x109
1.0 1.0 99.0 3.0x109