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Exercise 10 Antimicrobial Sensitivity Test

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28 views4 pages

Exercise 10 Antimicrobial Sensitivity Test

Uploaded by

Kent Tutor
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© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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TAGUM DOCTORS COLLEGE INC.

BSci 3L: Microbiology and Parasitology


Laboratory Manual

NAME: ______________________________________________ DATE: __________


COURSE & SECTION:__________________________________ GROUP NO.______

EXERCISE NO. 10

Antimicrobial Susceptibility Test

Antibiotics are given to treat infectious diseases. The physician faces the problem of
deciding which antibiotic to use for a given infectious disease. To make sure that the antibiotic
to be given is suited for a specific organism, an antimicrobial susceptibility test must be
requested. This test will tell the physician if the organism involved in the disease process is
susceptible to or resistant to a particular antibiotic, thereby saving the patient from spending
money on a drug that will not work on the particular organism involved in the first place.
Susceptibility testing is most often indicated when the etiologic agent involved is known
to be capable of developing resistance to commonly used antimicrobial agents. It is rarely done
if the organism is not known to develop resistance against a given antibiotics.

Materials:

Prepared culture of E. coli


Diffusion discs
Disinfectant or agents of your choice.
Commercially available antibiotics.
Ruler

PROCEDURES

Preparation of antibiotic disc


1. Prepare a Whatman number one (no 1) filter paper and punched to a disc size of 0.5cm
diameter using a standard office paper/file puncher.
2. Sterilize the discs by autoclaving before impregnation of antibiotics.
3. Dissolved the antibiotics in appropriate volumes of distilled water and dilute it.
4. Impregnate the disc by putting 1 ml of stock solution in a sterile plastic petri dish. The
sterilized filter paper discs were then added to the solution.
5. Allow to fully absorbed for 10 minutes and then dried in the oven at 40° C. Place the
prepared discs into labeled containers and store at 4° C until the testing.

Disk-diffusion assay:
1. On the culture plates, place equidistant from each other and in a circular fashion, one
disc each of the commercially prepared antibiotic discs. Incubate at 35 ° C for 16–18
hours.
2. After incubation, measure with a ruler the widest diameter of the zones of inhibition of
each antibiotic (expressed in millimeters).
3. Compare the measurements obtained with the reference table. Record your results with
the interpretation (susceptible, intermediate susceptibility, resistant) on the table on the
next page.
TAGUM DOCTORS COLLEGE INC.
BSci 3L: Microbiology and Parasitology
Laboratory Manual

RESULT SHEET

Agent Zone size Interpretation Photo


(mm)
TAGUM DOCTORS COLLEGE INC.
BSci 3L: Microbiology and Parasitology
Laboratory Manual

POST-LAB DISCUSSION:

1. What are the characteristics of an antimicrobial agent?

2. List 5 antimicrobial agents commonly used, indicate if it is antibiotics, antifungal, or


antiviral.

3. What is meant by resistance? Give the mechanisms by which organisms develop


resistance?

4. As future health professionals, why is it important to advocate for proper use of


antimicrobial drugs?
TAGUM DOCTORS COLLEGE INC.
BSci 3L: Microbiology and Parasitology
Laboratory Manual

Conclusion:

Reference:

Bartolome et al. 2020. Microbiology and Parasitology. A textbook and laboratory manual for the
health sciences. Second Edition.

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