Expt #11

Date: 8th March,2019.

Title: Planning and Designing #1- Alcohol concentration in infection prevention hand gels

Problem Statement: Many hospitals and other public buildings now use alcohol-based hand gels to help
prevent the spread of infection. These hand gels contain a high concentration of alcohol (such as
ethanol) and a mixture of other components to help thicken and perfume the gel. Plan and design an
investigation to discover the minimum concentration of ethanol that is needed to inhibit the growth of a
specific type of bacterium.

Introduction:

Streptococcus pyogenes (Group A streptococcus) is a bacterium that is extracellular, and it can cause
many diseases ranging from mild to deadly. A common infection caused by Group A streptococcus
bacterium is strep throat. This infection results in inflammation in the throat and it is found mainly in the
throat or skin. The greatest risk for this bacterium to spread is if a person meets someone who has strep
throat, encountering mucus from the nose or throat of an infected person or treating a wound or sore of
an infected person. Hence, a hospital without proper sanitation facilities, such as a hand gel with the
correct concentration of ethanol to kill this bacterium, can cause uninfected persons such as to contract
the bacteria

Hypothesis:

A hand gel with the concentration of 60% alcohol will be able to inhibit the growth of group A
streptococcus bacterium.

Aim: To plan and design an experiment to determine the minimum concentration of ethanol that is
needed to inhibit the growth of a specific bacterium.

Materials/ Apparatus:

100% ethanol, group A streptococcus bacterium, incubator, sterile petri dish, agar plate, six 6mm filter
paper disks, cotton swabs, 6 beakers, 20-microliter micropipette, measuring cylinder, distilled water,
sterile tweezer.

Variables

Controlled Variable: the amount of agar added to each petri dish

Independent Variable: The concentration of alcohol present in various filter paper disks

Dependent Variable: The radius of the zone of inhibition

Procedure:

1. Label six beakers: 0%, 20%, 40%, 60%, and 80%. Make 10cm 3 of each solution as stipulated in
the table below. Place the solutions in the respective beakers.
 Table 1: Dilution Series of ethanol

Concentration of ethanol/ % 0% 20% 40% 60% 80%

Volume of 100% ethanol/ cm3 0 20 40 60 80
3
Volume of distilled water/ cm 100 80 60 40 20

2. Place six 6mm filter paper disks into the sterile petri dish. Using the microliter micropipette,
measure 20 microliters of each solution, and place it on the filter paper disks.
3. Cover the petri dish and allow the filter paper disks to dry for 3 hours.
4. Mark off 5 equal sections on the agar plate. Label the sections with various concentrations of
ethanol prepared.
5. Using the microliter micropipette, measure 20 microliters of group A streptococcus bacterium
and add it to the agar plate. Spread the bacteria evenly on the plate.
6. Using a sterile tweezer, quickly place the dry filter paper disks in the center of the corresponding
section.
7. Place the agar plate in the incubator for 20 hours.
8. Measure and record the radius of the zone of inhibition.

Expected Results:

TABLE SHOWING THE RADIUS OF THE ZONE OF INHIBITION IN VARIOUS CONCENTRATIONS OF ETHANOL

Ethanol concentration in hand gel (%) The radius of the zone of inhibition (mm)
0
20
40
60
80

Treatment of Results:

Alcohols, such as ethanol, dissolve the phospholipid bilayer present in the bacteria’s cell surface
membrane. As a result, all organelles enclosed within the cell are leaked out causing the bacteria to die.
Since ethanol is dispensed from the filter paper disks, all the bacteria surrounding these disks will die
forming a zone of inhibition. The greater the radius of the zone of inhibition, that concentration of
ethanol is considered more effective.

To inhibit the growth and reproduction of bacteria, the ethanol concentration present within the hand
gels must be high. It is expected that the minimum concentration of ethanol required in a hand gel to
inhibit the growth of bacteria is 60%. This type of formula shows that the composition of hand gel is
majorly comprised of only alcohol, giving enough alcohol molecules to dissolve the cell surface
membrane present in the bacteria.
Precautions and Limitations/Assumptions

Assumption: It is assumed that agar present in the experiment will provide a natural environment for
bacterial growth, just as if it was on a surface area.

Precaution: There is a possibility of other types of bacteria within the surroundings of the conducted
experiment may alter the results, by reacting with the various concentrations of ethanol on the filter
paper disks. To minimize the occurrence of this error, the petri dish and the tweezers must be sterilized,
and the petri dish must be covered while the disks are drying.