Bacteria and Savlon
Aim: I want to investigate how the different concentration of Savlon can effect the growth and reproduction of Bacteria.
Hypothesis: I believe that the higher concentration of savlon the more effect it will have on the reproduction and growth of bacteria.
Fair test:
-Which variable will be changed? The concentration of the Savlon
-How will the independent variable be changed? By adding water to the solution
-Give a suitable range of values for this variable? There was 4 different values for the variable. for quadrant 1 there was 0% Savlon and 100% water. For quadrant 2 there was a dilution of 1/10. For quadrant 3 there was a dilution of 1/100, and finally for quadrant for there was a dilution of 1/1000.
-Which variable will have to be measured or observed in order to get some data or information from the investigation? The inhibition zone/ clear zone
-How will the independent variable be measured or observed? The variable will be measured by the diameter of the clear zone in mm.
Other variables that need to be controlled to make your results more accurate
Other variables
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Describe how this variable will be controlled or kept the same
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Temperature
| Storing the agar plate in the same room meaning the bacteria can grow in the same heat. |
Bacteria
| Making sure to use the same type of yoghurt on each plate |
The size of the disc
| Using the same cut hole punch to ensure the discs are the same size |
How will you ensure that your results are reliable?
We will ensure that our results are reliable by doing this experiment 3 times, and get an average result.
Equiptment:
1. Vivid
2. Agar plate
3. Savlon
4. Cotton bud
5. Yoghurt
6. Savlon
7. Water
8. Hole punch
9. Filter paper the
Method:
1. Using a vivid you need to draw two lines on the agar plate which splits it into four even areas.
2. We then get a cotton bud and dip it into the beaker of yogurt and then spread it evenly on the agar plate, and then shut the lid to ensure that the plate stays sterile.
3. Using a plastic pipette you want to create your dilutions.
-1 drop of Savlon to 9 drops of water (1/10 dilution)
-1 drop of mixture above to 9 drops of water (1/100 dilution)
-1 drop of mixture above to 9 drops of water (1/1000 dilution)
4. Each dilution is placed in a spot of the spotting tray
5. Get filter paper and cut out 4 small holes with a hole punch
6. Dip the tweezers in the jar of Ethanol to sterilize them and pass through the flame of the Bunsen Burner.
-With this step make sure the lid of the Ethanol jar is closed at all times apart from when you are dipping in the tweezers.
7. With tweezers pick up each hole punched piece of filter paper and dip it in each concentration of savlon.

- 1. The control/Neutral-water
- 2. 1/10 dilution
- 3. 1/100 dilution
- 4. 1/1000 dilution
8. Place the disc on a paper towel to remove excess savlon solution.
9. Place each disc in the center of the quadrant.
10. Once all discs are placed on the agar plate then shut the lid and sellotape it shut.
11. Get your agar plate, and store in a warm place overnight.
12. Repeat this process with 3 agar plates to ensure and even result.
Change in method: We changed step 2 of the method. Instead of using a cotton bud to apply the yogurt on to the agar plate we decided to water the yogurt down and pour a thin layer onto the agar plate and swish it around until the plate was completely covered. We changed this step to decrease the chances of contamination.
Data
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Concentration of the savlon
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The clear zone (mm)
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Average diameter
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|
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Plate 1
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Plate 2
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Concentration 1
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Control
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0 mm
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0 mm
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0 mm
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Concentration 2
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10%
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13 mm
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20 mm
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16.5 mm
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Concentration 3
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1%
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9 mm
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10 mm
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9.5 mm
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Concentration 4
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0.1%
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5 mm
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6mm
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5.5 mm
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Diameter of Clear Zone Around Different Concentration Of Savlon.
Conclusion: In conclusion my hypothesis was correct. The higher concentration of savlon had a bigger effect on the growth and reproduction of Bacteria. After looking at the graph we can see that there is an increase in the trend line with an increase in the concentration of Savlon. There is a larger diameter around the disc where there there is a higher concentration of the savlon which is how we can see the effects on the growth and reproduction.
Discussion and evaluation of the method and data:
Savlon is an antiseptic used to help infections and clean wounds, cuts, burns, grazes and stings. The active ingredient in Savlon is Cetrimide
C19H42BrN and Chlorhexidine
C22H30Cl2N10. When cetrimide comes in contact with bacteria it inhibits the bacteria within 15 seconds. Both Cetrimide and chlorhexidine kills bacteria because they have a positive charged molecule and bacteria has a negative phospholipids in the cell wall. This would be considered active transport. Active transport is when there is a movement of molecules across the cell membrane from a lower concentration to a higher concentration. This causes a rupture in the cell wall leading to the chlorhexidine fluid to leak into the inside of the cell and attacks the cytoplasmic membrane which leads to the death of the bacteria.
Bacteria is a single cellular micro-organism. On the outside it has a layer called a capsule, it works as defence against immune systems. The inner layer is called a well wall, this is a strong layer that protects and provides shape of the cell. Next we have the cell membrane which is a flexible barrier that works as a control of what enters and exits the cell. Inside the cell we have the cytoplasm which holds all the cell organelles such as a nucleotide which is the genetic information of the cell, we also have two small circular pieces of DNA which codes for essential processes of the bacteria called plasmids, and ribosomes which is the location of protein synthesis in the cell structure of bacteria. On the outside of the cell we have many pili's which works as a bridge to connect to other bacterias and allow them to grow. We also have a flagellum which is the tail of that cell that aids the cell in movement. Bacteria replicate asexually, this means they are genetically identical to each other, although they can still mutate. Bacteria also replicate using Binary Fission. Binary fission is when a single cell divides into two identical daughter cells. With this form of reproduction bacteria then start to form colonies which contains thousands of bacteria. We clearly could start to see the bacteria on the agar plate when the colonies had formed. Bacteria has many phases it goes through when it comes to growth. The first phase is the lag phase, this is when the bacteria is initially placed in the culture they begin to take the nutrients and synthesise their RNA. Phase 2 is the exponential phase, this is the phase when there is a huge spike in bacteria reproduction. Phase 3 is the stationary phase when the bacteria growth and death rate are equal due to a reduction of nutrients. The last phase, phase 4 is the death phase which is when there is a drop of numbers of bacteria due to the bacteria dying from lack of nutrients. With the Cetrimide and Chlorhexidine in the Savlon on the discs they would have killed the bacteria and stopped the growth in a particular area which we would consider the clear zone, or the zone of inhibition. MRS GREN is an acronym that is used to remember the life process of living organisms. It stands for Movement, respiration, sensitivity, growth, reproduction, excretion, nutrition. Bacteria mainly use growth and reproduction.
Evaluation:
Overall our experiment i would say was not reliable. I believe this is because only two out of three of our agar plates had reliable data.We were unable to measure the diameter around the discs on plate three, this is how it was unreliable, so we chose not include plate three's data in our graph. We also chose to change step 2 of our method to lower the risk of contamination. We poured a thin amount watered down yogurt on the agar plate instead of swabbing yogurt onto it with a cotton bud. If we were to do this experiment again i would chose a different range of variables making the concentration of savlon higher so there would be a larger clear zone to measure. The bacteria in the yogurt we used was Streptococcus thermophilus and lactobacillus bulgaricus. Streptococcus thermophilus is a probiotic strain of bacteria. It is used to break down sugar in milk. Lactobacillus bulgaricus is one of the healthy bacteria, which works to keep the pH scale lower in the gastrointestinal tract so that bad bacteria can not take a hold and colonize. For our experiment we used nutrient agar which is a general purpose medium that contains 0.5% peptone, 0.3%yeast extract, 1.5% agar, 0.5% sodium chloride and water. For our experiment the agar plates were kept in an incubator at a temperature of 25 degrees celsius. If the incubator was set at a higher temperature the bacteria would get a chance to grow more rapidly.
Bibliography
http://www.wired.co.uk/article/whats-inside-savlon-antiseptic-cream
http://chlorhexidinefacts.com/mechanism-of-action.html
https://nootriment.com/lactobacillus-bulgaricus-streptococcus-thermophilus/