Adventist University of the Philippines

COLLEGE OF SCIENCE AND TECHNOLOGY

Department of Biology
KIRBY BAUER METHOD

Name: Maureen Joy V. De Guzman

The Kirby-Bauer disk diffusion susceptibility test determines the sensitivity or resistance of pathogenic
bacteria to various antimicrobial compounds in order to assist physicians in selecting treatment options
their patients.

Objectives:

1. Determine the procedure for the Kirby Bauer method.

2. Identify the results for the Kirby Bauer method.
3. Interpret the results for any given antibiotic tests

Instructions

1. After the lecture discussion, watch the procedure for the Kirby Bauer test. (152) Kirby Bauer Disk
Diffusion Method - YouTube
2. (152) Microbiology resources - How to Measure Zones of Inhibition (ZOI) - YouTube

Answer the following:

1. Identify the agar used for the Kirby Bauer method? Mueller-Hinton agar
2. Identify the instrument for evenly distributing the bacterial smear? Sterile swab
3. Identify the incubation period for doing the Kirby Bauer Test? 18-24 hours
4. Compare the following terms:

Susceptible Intermediate Resistant

Can be treated with the drug May be treatable with the Cannot be treated with the
drug but may require adjusted drug
dosage.

5. Describe the term ZONE OF INHIBITION.

The bacterium is swabbed on agar, then antibiotic discs are placed on top. The antibiotic diffuses
into the agar in diminishing proportions as one moves away from the disc. If the antibiotic
concentration kills or inhibits the organism, there will be no growth in the immediate area around
the disc: this is known as the zone of inhibition.
Interpret the following data:

1. Escherichia coli with 12 mm using Cefazolin: Resistant

2. Bacillus subtilis with 26 mm using Clindamycin: Susceptible
3. Staphylococcus aureus with 17 mm using Erythromycin: Intermediate
4. Streptococcus sanguine with 19 mm using Gentamicin: Susceptible
5. Klebsiella pneumonia with 9 mm using Tobramycin: Resistant

Which better to use: Broad spectrum antibiotics or Narrow spectrum antibiotics?

Narrow-spectrum antibiotics are only effective against a few species of bacteria. Broad-
spectrum antibiotics are effective against a wide range of microorganisms. Both forms are effective at
treating infections. However, administering broad-spectrum antibiotics when they are not required can
result in antibiotic-resistant bacteria that are difficult to cure.

How come bacteria becomes resistant?

Through mutation and selection, bacteria can develop defensive mechanisms against antibiotics.
For instance, some bacteria have created biochemical “pumps” that may eliminate an antibiotic before
it reaches its target, while others have evolved to manufacture enzymes that deactivate the antibiotic.
Antibiotic resistance is increased by overuse and misuse of antibiotics, as well as poor infection control
and prevention.