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Practical 6.3 Which antibiotic is most effective?

Purpose Safety

• To investigate the effect of
different antibiotics on bacteria.
Eye protection is not necessarily required during microbial
work, though it is good standard practice.
The microorganisms are a potential biological hazard. Use
aseptic techniques when transferring the bacteria to the Petri
dishes. Clean the bench with antibacterial disinfectant. Do
NOT open the Petri dishes once they have been incubated.

Notes on the procedure

The aim of the practical is to show students a method of determining bacterial sensitivity to
different antibiotics and to give them practice at using aseptic techniques. This could be done
with antibiotic discs or the same technique could be used with antiseptics. The Student sheet
suggests using different bacterial species but the activity could be done using just one species.
However, using different species should illustrate that they are not all equally susceptible.

Although allergic response by patients is considered in the questions, where antibiotics are
already impregnated on to discs, the risk for students with allergic responses is not great.
Discs are not handled directly and no airborne dust is created by the discs.

The behaviour of students must be considered. If they might attempt to lift a piece of tape
from an incubated dish, the agar plates should be sealed around the circumference after
incubation but before being returned for observation. Also, to prevent agar plates being
removed from the lab, count all plates back in again before students are allowed to leave.

Answers
1 Choice of strategy may vary, but it must be clear and easy to carry out and produce
reliable results.

2 The rate of diffusion of the antibiotic will be influenced by the size of molecule, its
concentration and the potency of the antibiotic. If the antibiotic is effective at lower
concentrations the circle will be larger (all other things being equal). Gram-positive and
Gram-negative bacteria respond differently to antibiotics.

3 Responses will depend on the data, but should show a clear understanding of the
concepts of accuracy, validity and/or reliability when explaining variation. Suggestions
for the presentation of variation in graphs may also vary but need to identify anomalous
results clearly so that conclusion are obviously based on reliable results.

4 You may have to consider whether the patient is allergic to any of the antibiotics. For
example, allergy to penicillin is not uncommon. You may consider the state of the patient’s
immune system. In patients with a weakened immune system you would not want to
use a bacteriostatic antibiotic, that is, one that stops bacterial reproduction but does not
kill the bacteria. Some antibiotics can be used together and produce a larger effect when
combined than if administered separately. This is known as synergism.

Edexcel practical materials created by Salters-Nuffield Advanced Biology, ©University of York Science Education Group.

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