Carl Andrew G.

Alvero
4, 3C-MT
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?
To determine which drug is more efficient against the bacteria used compared to the other drugs, If those
criteria should be different, how would it possible to determine which drug is actually more efficient against
certain bacterium?

2. Why are E. coli, P. aeroginosa, and S. aureus used as standards in the Kirby Bauer method?
Please include your references.
Recommended organisms for quality assurance purposes are Staphylococcus aureus ATCC 25923
(Biosafety level (BSL) 2), Escherichia coli ATCC 25922 (BSL 1), and Pseudomonas aeruginosa ATCC
27853 (BSL 2) (www.atcc.org), as the zone of inhibition for these organisms is known. Because the zone
sizes are known for these organisms, they are recommended for use in the educational setting, although
the use of unknowns should also be incorporated into the educational experience. For quality control
testing, the zone sizes for these three organisms can be found on the package insert from any
antimicrobial disk you purchase.
From: http://www.microbelibrary.org/component/resource/laboratory-test/3189-kirby-bauer-disk-diffusionsusceptibility-test-protocol
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."

Approximate Cell Count Density (x108 cells)

0.6
0.7
0.8
0.9
1.0

McFarland

0.1

0.2

0.3

0.4

0.5

standard no.
1.0% Barium

0.01

0.02

0.03

0.04

0.05

0.06

0.07

0.08

0.09

0.1

chloride (ml)
1.0% Sulfuric

9.99

9.98

9.97

9.96

9.95

9.94

9.93

9.92

9.91

9.9

acid (ml)
McFarland Standard
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0

0.3
0.6
0.9
1.2
1.5
1.8
2.1
2.4
2.7
3.0