Effect of antibiotics on bacteria

affect of penicillin on micrococcus luteus which is Gram +ve, and varied the strength of the
penicillin disks. I had to do several repeats at each strength,
Aim:
My aim is to investigate the effect of different concentrations of antibiotics on the growth of
bacteria. The aim of this investigation is to find out what effect out of two antibiotics, penicillin
and streptomycin has on the growth and multiplication of two different species of bacteria.
The two different types of bacteria we will be using are E-coli and micrococcus luteus.
Hypothesis:
I predict that as the concentration of the antibiotics increases the bacteria growth decreases
Null Hypothesis:
There is no link between the concentration of antibiotics and the effects it has on the growth
of the bacteria
Theory:
Antibiotics work in 4 ways, which are listed below:
1. Cell Membrane Disruption - This involves making the cell fully permeable which results in
substances moving into it causing it to burst and so kills the bacteria
2. Inhibiting Nucleic Acid Synthesis - This method doesn't kill the bacteria off but keeps the
growth level static. The bacteria isn't able to replicate its DNA and so no binary fission occurs,
causing the growth level to become static
3. Inhibiting Cell Wall Synthesis - This is where an antibiotic inhibits the enzyme required to
form cross links within the cell wall and as a result the bacteria looses its structure and is
unable to function properly .
4. Inhibiting mRNA Translation - This is where the Translation part of protein synthesis is
inhibited by binding across the bacterial ribosome meaning proteins and enzymes the
bacteria it requires isn't made and so dies.
Therefore, it is logical to presume that the higher the concentration of antibiotics the more
effective it will be in wiping out, and killing the bacteria. Because the more antibiotic
molecules there is in the solution means there is a higher chance that these antibiotic
molecules will come upon a bacterial cell and disrupt the cell and kill it . So in conclusion it is
logical to state that the higher the concentration of antibiotic the more effective the antibiotic
will be at killing of the bacteria.
Dependant Variable:
There are a number of
different ways of measuring the effectiveness of the antibiotics but I will be using the
technique of measuring the zone of inhibition. This method provides data which can easily be
measured.
The zone of inhibition (of killed bacteria) in mm (+/- 0.5mm). Measured with a ruler.

Independent Variable
For me to fully determine the effectiveness of the antibiotic there will have to be an
independent variable, which is the concentration of the antibiotic. So I have decided that I am
going to use a wide degree of concentrations of antibiotic and the gap between the
concentrations needs to be kept to a minimum cause the optimum concentration may be left
out if the gaps are too big. Also because the results will be analyzed by the spearman's rank
correlation I will use 7 data values.
Apparatus:
* Four sterilised Agar plates, * Glass spreader, * Masking tape, * Wax pencil, * Lab roll, * A
beaker with 70% alcohol, * E- coli (Culture) * Micrococcus luteus (culture), * Penicillin
(antibiotics), * Streptomycin (antibiotics). * 1 x metal forceps * 1 x Bunsen burner and mat * 1
x scissors * Matches * Sellotape *

Results:
The following is a table to show how large the zone of inhibition for each antibiotic is in mm.
The Petri dishes were left for 24 hours at a temperature of 30C. (+/-0.5C). Different
antibiotics Zone of inhibition/ mm (+/- 0.5mm)

Risk Assessment
Glassware: Dangerous if glass is broken. Low risk and perfectly safe if used properly. Do not
try to pick up pieces of broken glass with your hands, especially from a wet sink. Report all
breakages immediately so that staff can deal with it. Inform all students of any broken glass
so they move away from it and avoid injury. Inform staff so they can deal with it. Seek medical
attention.
Trip Hazards: Bags, coats, chairs. Low risk if bags, coats and chairs are put or stored in a
safe area. Put baggage etc well under benches, away from the working area or store at the
front of lab. Inform staff and seek medical attention. Help the person if possible.
Bunsen Burner: Could burn the skin and can set clothing on fire. Low risk if used correctly
and safely. Pull lever down when Bunsen burner is not in use, this reduces the intensity of the
flame so contact of flame with objects is less likely, wear safety glasses and lab coat. Break
fire alarm, seek medical attention, and wash with cold water. Escherichia coli: Can cause
food poisoning, diarrhoea and infections can be fatal. Low risk because low concentration is
used. Low risk if followed the method carefully. Wear safety goggles, gloves and lab coat. Tell
staff and seek medical help immediately.
Bacillus subtilis: Can cause severe eye infections Low risk because low concentration is
used. Low risk if followed the method carefully. Wear safety goggles, gloves and lab coat. Tell
staff and seek medical help immediately.
Controlling variables:

To be sure that only the antibiotic is responsible for the clear zone that will be present
in the Petri dish all other variables must be kept constant. I will do this by:
Volume - The total volume of the bacteria, buffer and water solution will be 6cm3 .This
will eliminate the volume becoming a variable which may give erroneous results.
Surface area of Agar - I will keep this constant by using Petri dishes all the same sizes.
This will prevent surface area becoming a variable cause having a higher surface area
may affect the rate at which the bacteria are killed. Also the same to keep the size of
clear zone to be measured the same
Surface area of Wells - I will keep this at 5mm, to prevent this becoming a variable.
Temperature - all dishes are in the same incubator. Amount of nutrient available for
bacterial growth - use the same amount of agar for each.
E.coli was used for both dishes to keep the strain of bacteria same.
Sterilised syringes, forceps and loop are used for both dishes, so unwanted
contamination is avoided.