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Disk Diffusion Test (Kirby- Bauer Disk Diffusion)

Proteus vulgaris
Otani, Aki S.
MT10315

INTRODUCTION

Proteus vulgaris is a rod-shaped Gram-negative chemoheterotroph bacterium. The
size of individual cells varies from 0.4~0.6m by 1.2~2.5m. P. vulgaris possesses peritrichous
flagella, making it actively motile. It inhabits the soil, polluted water, raw
meat, gastrointestinaltracts of animals, and dust. In humans, Proteus species most frequently
cause urinary tract infections, but can also produce severe abscesses;P. mirabilis produces 90
percent of cases, and is encountered in the community, but P. vulgaris is associated
with nosocomial infection. (Citizendium, 2010)
Proteus vulgaris, a motile organism, in a sulfide indole motility (SIM) deep. The black
precipitate indicates hydrogen sulfide production by the organism. SIM medium is used to test
for hydrogen sulfide production, indole production, and motility. An uninoculated SIM deep is a
transparent yellow semisolid (3 g/liter of agar) medium. Inoculate the SIM deep with an
inoculating needle, making the stab perpendicular to the agar surface. (ASM, 2011)
The Kirby-Bauer test for antibiotic susceptibility, called the disc diffusion test, is a
standard that as been used for years. First developed in the 1950s, it was refined and by W.
Kirby and A. Bauer, then standardized by the World Health Organization in 1961. It has been
superseded in clinical labs by automated tests. But the K-B is still used in some labs, or used
with certain bacteria that automation does not work well with. This test is used to determine the
resistance or sensitivity of aerobes or facultative anaerobes to specific chemicals, which can
then be used by the clinician for treatment of patients with bacterial infections. The presence or
absence of an inhibitory area around the disc identifies the bacterial sensitivity to the drug. The
basics are easy: The bacterium is swabbed on the agar and the antibiotic discs are placed on
top. The antibiotic diffuses from the disc into the agar in decreasing amounts the further it is
away from the disc. If the organism is killed or inhibited by the concentration of the antibiotic,
there will be NO growth in the immediate area around the disc: This is called the zone of
inhibition. The zone sizes are looked up on a standardized chart to give a result of sensitive,
resistant, or intermediate. Many charts have a corresponding column that also gives the MIC
(minimal inhibitory concentration) for that drug. The MIC is currently the standard test run for
antibiotic sensitivity testing because it produces more pertinent information on minimal dosages.
The Mueller-Hinton medium being used for the Kirby-Bauer test is very high in protein.

RESULTS AND DISCUSSION

Figure 1 shows the resulting MHA
plate that was incubated for 24 hours under
body temperature (37.5C). Proteus vulgaris
was the organism used and the antibiotic
disks that were inoculated are Vancomycin,
Kanamycin, Ampicilin and Oxacilin which
yielded almost similar result except for
Kanamycin which show a zone of inhinition.


Figure 1: Shows the resulting MHA plate that was cultured adn
incubated for 24 hours. K (Kanamycin) is the only antibiotic that is
susceptible to the organism.
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Figure 2 is the picture of the zone inhibited by the
Kanamycin disk. The zone of inhibition is 2.6 cm or 26 mm. It
is therefore Resistant to the organism used based on the
table 1.







Figure 3 shows no signs of inhibition due to the presence of colonies surrounding the antibiotic
disks. Therefore, Vancomycin, Oxacilin and Ampicilin are not suitable for treating Proteus
vulgaris infections.





Results of the measured
zone of inhibition was based
on this chart.



Figure 2 Shows Kanamycin and its zone of
inhibition in the MHA plate.
Figure 3 Shows the three susceptible antibiotics which portray no sign of inhibition.
Table 1 Shows the Lists of Antibiotics and the basis of inhibition as Susceptiple, Resistant or
Intermediate.
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Table 2. Diffusion Test Results
Proteus vulgaris
Drug Used Oxacilin Kanamycin Ampicilin Vancomycin
Result
Resistant Susceptible Resistant Resistant
Zone of
Inhibition
- 26mm - -

Table 2 shows the tabulation of yielded results. Oxacilin, Ampicilin and Vancomycin are
all resistant and Kanamycin is the only antibiotic that yielded a zone of inhibition with the
diameter of 26mm which makes it susceptible according to Table 1.
I therefore conclude that based on the results of the experiment done using the
organism, Proteus vulgaris, Kanamycin is the only antibiotic capable of inhibiting the said
organism among the other three antibiotic disks used thus Kanamycin may be used as an
antibiotic drug against Proteus vulgaris infections.

REFERENCES

ASM. (2011, NOvember 1). Proteus vulgaris, a Motile Organism, in a Sulfide Indole Motility Deep.
Retrieved September 30, 2012, from American Society for Microbiology:
http://www.microbelibrary.org/library/2-associated-figure-resource/3643-proteus-vulgaris-a-motile-
organism-in-a-sim-deep
Citizendium. (2010, December). Proteus vulgaris. Retrieved September 30, 2012, from Citizendium:
http://en.citizendium.org/wiki/proteus_vulgaris#References