A - Group research proposal




Investigative Skills in Science

Research Proposal Form
Project Title A: Investigation of the effect of different chemicals on the growth of bacteria
Project Title B: Investigation of the effect of different concentrations of Clorox on the growth of bacteria
Class
S2-08
Group: 
B
SN
Name of student
Email 
1
Chia Howie
chia_howie@s2019.ssts.edu.sg
2
Sung Yeji
sung_yeji@s2019.ssts.edu.sg
3
Joel Lim 
lim_xuan_wei_joel@s2019.ssts.edu.sg
Type of research: Observational and exploratory research
X
Test a hypothesis: Hypothesis-driven research
e.g. Investigation of the antibacterial effect of chrysanthemum

Measure a value: Experimental research (I)
e.g. Determination of the mass of Jupiter using planetary photography
X
Measure a function or relationship: Experimental research (II)
e.g. Investigation of the effect of temperature on the growth of crystals

Construct a model: Theoretical sciences and applied mathematics
e.g. Modeling of the cooling curve of naphthalene 

Observational and exploratory research
e.g. Investigation of the soil quality in SST
Category of research:
Sub-category: 
Biology
Microbiology
Reference 



Research Plan
Project Title A: Investigation of the effect of different chemicals on the growth of bacteria
Project Title B: Investigation of the effect of different concentrations of Clorox on the growth
of bacteria
1. RATIONALE: It includes a brief synopsis of the background that supports your research problem and explain why
this research is important and if applicable, explain any societal impact of your research. Include at least 3 in-text citations in your rationale. (500 words)  
Experiment A
Bacteria is a member of a large group of unicellular microorganisms which have cell walls but lack organelles and an organized nucleus, including some which can cause disease (Lexico, 2020). While most microorganisms are harmless, some species of bacteria cause death. The WHO states that "Inadequate sanitation is estimated to cause 432 000 diarrhoeal deaths annually and is a major factor in several neglected tropical diseases, including
intestinal worms, schistosomiasis, and trachoma. Poor sanitation also contributes to malnutrition. Some 827 000 people in low- and middle-income countries die as a result of inadequate water, sanitation, and hygiene each year, representing 60% of total diarrhoeal deaths. It is believed that poor sanitation is the main cause in some 432 000 of these deaths." (World Health Organisation 2019). 
Related to bacteria, cleanliness is the state or quality of being clean or being kept clean (Lexico, 2020)For school cleaning and disinfection, multiple challenges exist. First, although 'clean' may be generally defined based on the absence of visible soil, infectious pathogens cannot be seen or easily measured in the home. As a result, one cannot readily discern to what extent disinfection is needed or achieved. Second, the resident or cleaner may know nothing about pathogens other than a vague sense of the need for cleanliness and disinfection. They may not know or understand the difference between cleaning and disinfection,
including the fact that a surface should be cleaned before it is disinfected. (Goodyear, N., 2015) 
According to research, in 2017, 45% of the global population (3.4 billion people) used safely managed sanitation service. Good hygiene can help your psychological health.
Good hygiene is crucial to good overall health and wellness because it helps lower the risk for disease, illness, and medical conditions caused by the effects of poor hygiene.
When a person doesn't practise good hygiene, their body can accumulate bacteria that contribute to diseases such as athlete's foot, head lice, and scabies. Poor health can
also cause parasites to grow and multiply on the skin and in the body, causing parasitic diseases like malaria and toxoplasmosis (Mayo Clinic, October 03, 2017)
Good hygiene lowers your risk for infections and illnesses commonly spread through viruses and bacteria. Your hands come into contact with bacteria every time you cough,
use the restroom, touch your pet, or touch surfaces, such as stair railings, frequently used by others. Failing to protect your hygiene allows illness-causing bacteria into your body,
increasing the risk for disease.
There are no risks to practising good hygiene. Poor hygiene, on the other hand, increases the risk for bacterial, viral, and parasitic infections. Severe medical conditions that can
develop on behalf of poor health include gastroenteritis (WebMD, 2020), food poisoning, hepatitis A, influenza, (Solvhealth, 2020) common cold, (Solvhealth, 2020) giardiasis (Healthline, 2020), roundworm, and threadworm. Good hygiene can help you avoid the possible risks associated with poor sanitation. (Solvhealth, 2020) Using chemicals, such as alcohol ethoxylate or chlorine, can kill almost every bacteria. Products labelled "antibacterial" contain additional bacteria-killing substances such as triclosan (U.S. Food & Drug Administration, 2020) or triclocarban. (Wikipedia, December 16 2019) For example, Jif contains sodium carbonate, sodium dodecylbenzene sulfonate and calcium carbonate kill bacteria. (Unilever, 2020)
Hence our idea, "Investigation of the effect of different chemicals on the growth of bacteria" is to benefit our knowledge and to provide a cleaner environment for SST students. 
This investigation is essential as we can apply it to our everyday life in school. We can even improve the whole world's sanitation. Sanitation is vital to every human as it is healthy. Adequate sanitation means we can reduce the risk of infecting diseases to our body. 
So, by carrying out this experiment, we would be able to confirm which chemicals are active in killing bacteria are most effective in killing the germs in our everyday lives. From there, we
hope to improve the school and the world's sanitation by finding out which soaps are the most efficient. This will help reduce the number of illnesses and deaths linked to unsanitary environments
and may help improve other companies soaps. Companies could also make a cheaper version, helping third world countries like Africa, where inadequate sanitization is frequent.
Experiment B
Clorox is a type of bleach from The Clorox Company. Household bleaches are a part of everyday life and are in nearly every home, usually in more than one form. There are two main classes
of household bleach: chlorine bleaches and non-chlorine bleaches. All of these bleaches are in a class of chemicals known as oxidative agents, meaning that they cause a chemical reaction
called oxidation when they come into contact with certain stains, certain germs or other organisms, and sometimes clothing dyes.

Bleach is a very useful chemical both around the house and for large-scale use. You can use bleach to remove stains on clothing or to whiten your laundry. It's used to disinfect surfaces, too, especially in the kitchen and bathroom. Hospital personnel use bleach as a disinfectant, hotels use bleach to clean and disinfect bed linens and surfaces, and restaurants disinfect food preparation surfaces with chlorine bleach. People use chlorine in swimming pools to keep the water clean and raise the pH, and in much smaller concentrations to help keep municipal water supplies free of
harmful organisms. Companies sometimes add chlorine bleach to industrial wastewater to reduce odor, and chlorine is used by the glass, chemical, pharmaceutical, textile, agriculture, paint and paper industries.
With its many uses, bleach is a very familiar product to most people. (Melissa, S, 2009) Clorox is a chlorine bleach. Chlorine bleach contains the active ingredient sodium hypochlorite (NaOCl), while non-chlorine bleaches have different active ingredients for different purposes.
Sodium Hypochlorite performs the bleaching, stain removal and disinfection functions. This chemical is highly effective in killing bacteria. Since Clorox is made of this chemical, this proves that Clorox is good in killing bacteria. Chlorine bleach kills Vibrio cholerae, the bacterium that causes cholera, a disease that killed in epidemic proportions before water treatment. It can still kill in countries where clean drinking water is not available. Chlorine bleach can also kill dangerous bacteria and viruses on surfaces, such as methicillin-resistant Staphylococcus aureus, influenza and HIV. Chlorine bleach is especially valuable as a disinfectant since germs are not able to develop immunity against it, as they have done against certain drugs (Lenntech, 2020).  
To kill germs, sodium hypochlorite uses the same quality that makes it such a great stain remover -- its power as an oxidizing agent. When sodium hypochlorite comes in contact with viruses, bacteria, mold or fungi, it oxidizes molecules in the cells of the germs and kills them. Scientists also believe that the hypochlorous acid that forms when sodium hypochlorite is added to water can break down the cell walls of some germs. The hypochlorous acid also seems to be able to cause certain proteins to build up in bacteria, making their cells unable to function (Winter, J, 2008). Non-chlorine bleaches that are oxidizing agents can also act as
disinfectants on some surfaces, but they are less potent than chlorine bleach. Chlorine bleach, when used properly, is a practical and effective disinfectant. This shows that Chlorine is highly effective in killing bacteria,
especially viruses even like the recent 2019 coronavirus. 
The Environmental Protection Agency (EPA) has evaluated multiple scientific studies on the effects of chlorinated drinking water, and the organization's found no evidence of risk for cancer, reproductive problems or birth defects. The European Commission (EC) also determined that the most common sources of exposure to chlorine bleach is through skin contact when using bleach for cleaning at home or through ingestion of chlorinated drinking water. Swallowing small amounts of swimming pool water may also be a risk, but there is no significant indirect exposure through the environment. The Commission determined that there is no
evidence of negative health effects due to long-term exposure to small amounts of chlorine bleach. These results show that Clorox is safe to use. (Melissa, S, 2009)
Further research in this experiment can allow us to determine the amount of concentration needed from the Clorox to kill bacteria. This will allow us to cut cost by using a bottle for a longer time. We would also be able to prove that a lower concentration of Clorox is as effective as the undiluted Clorox in killing viruses like the 2019 coronavirus on surfaces. 
2. RESEARCH QUESTION(S):
2.1 Research question being addressed 
What are the effects of different chemicals like Mama lemon and Mr Muscle and more on the growth of canteen table bacteria?
2.2 Hypotheses
Experiment A:
The hypothesis is the more acidic the chemical is, the less likely bacteria will survive. 
Experiment B:
The hypothesis is the less concentrated Clorox is equally as effective as the fully concentrated Clorox in killing bacteria. 
2.2.1 Independent variable
The type of cleaning product tested to kill the bacteria
2.2.2 Dependent variable 
The diameter of the inhibition of bacterial growth
2.2.3 Controlled variables
A)
  1. source of bacteria used
  2. time given for the bacteria to grow
  3. mass of chemical used
B)
  1. the volume of dilute used
  2. the time given for bacteria to grow
  3. source of bacteria used
2.3 How is this based on the rationale described above
A. If the hypothesis is correct, this science project can be used to further the research by finding out which soaps are the most efficient. B. It is to benefit our knowledge and to provide a cleaner environment for SST students. Also, this will help reduce the number of illnesses and deaths linked to unsanitary environments and may help improve other companies soaps. Companies could also make a cheaper version, helping third world countries like Africa, where inadequate sanitation is frequent. This experiment can improve the school and the world’s sanitation. C. If the hypothesis is correct, this experiment can allow us to determine the amount of concentration needed from the Clorox to kill bacteria. This will allow us to cut cost by using a bottle for a longer time. We would also be able to prove that a lower concentration of Clorox is as effective as the undiluted Clorox in killing viruses like the 2019 coronavirus on surfaces.
3.    Method
3.1 Equipment list
Experiment A:
Items:
Quantity:

Sterilised paper disk
5
Petri dishes 
6
Samples of bacteria in canteen 
0.3ml in total, 0.05ml / 50 ul for each petri dish
Labels for petri dishes 
6 post-its labelled
Mama Lemon
1ml 
Clorox 
1ml 
Dettol 
1ml 
Jif 
1ml 
Mr Muscle
1ml 
Safety materials (goggles, gloves, masks) 
x3 sets
Parafilm 
1 strip for each petri dish, 6 strips in total
Agar Jelly 
75 ml in total, 12.5 ml for each petri dish
5ml small bottles 
5 bottles, each one to be filled with 1ml of each respective soap. 
15cm ruler
1
Cotton bud swab
1
Tweezer
1
Experiment B:
Items:
Quantity:

Sterilised paper disk
5
Petri dishes 
6
Samples of bacteria in canteen 
0.3ml in total, 0.05ml / 50 ul for each petri dish
Labels for petri dishes 
6 post-its labelled
Sterilised water
1ml 
Clorox 
1ml 
Test tube
5
Test tube holder
1
Safety materials (goggles, gloves, masks) 
x3 sets
Parafilm 
1 strip for each petri dish, 6 strips in total
Agar Jelly 
75 ml in total, 12.5 ml for each petri dish
5ml small bottles 
5 bottles, each one to be filled with 1ml of each respective soap. 
15cm ruler
1
Cotton bud swab
1
Tweezer
1
3.2 Diagrams
Experiment A  


Figure 1: Set-up procedure to prevent dust from entering the petri dishes. 
Figure 2: Set-up procedure on how to soak and place a paper disk into a 
petri dish.



 Figure 3: Experimental setup used to measure the killzone of the bacteria caused by
different killing agents in each petri dish.  

Experiment B


Figure 1: Set-up procedure to show where the different concentrations of Clorox should be placed. 



Figure 2: Set-up procedure on how to soak and place a paper disk into a petri dish. 



    Figure 3: Experimental setup used to measure the kill zone of the bacteria caused by different

concentrations of Clorox in each petri dish. 
3.3 Procedures: Detail all procedures and experimental design to be used for data collection
Experiment A

According to Tom Besty, DC and Jim Keogh, RN, (2005), step 6 was adapted from their book,
Microbiology Demystified from chapter 6, page 104 - 105. 
According to Wayne, C, (2015), steps
1 - 4, 6 - 13 and 15-18 were adapted from their website, ISS S205 Group A, Annex A, Group
project proposal. 

According to Meredith, J, (2019), step 14 was adapted from their website, Wikihow, How to grow bacteria in a Petri dish. 

  1. Collect sample bacteria from the canteen uncle. 
  2. Set up 6 Petri dishes.
  3. Light up the alcohol lamp to create an updraft to prevent dust from entering the Petri dishes. 
  4. Put 12.5 ml of agar jelly into each petri dish.
  5. Use a cotton bud swab to spread 50ul of sample bacteria evenly from school canteen into each of the Petri dishes. Only put on safety goggles at this point to prevent bacteria from getting into the eye.  
  6. Leave 17.45ml as bacteria need carbon dioxide and oxygen to live. 
  7. Use a tweezer and soak each sterilised filter paper disk into each solution (Mama lemon, Clorox, Dettol, Jif and Mr Muscle)
  8. Soak a total of 5 sterilised filter paper disks of different solutions. 
  9. Place one different soaked sterilised filter paper disk into each petri dish. 
  10. Do not place any sterilised filter paper disk for the last one as it is the control variable. 
  11. Sterilise the tweezer after placing a soaked sterilised filter paper disk by putting above the alcohol lamp to prevent the different soaps from mixing. 
  12. Name all the different Petri dishes with a sticker to a label. 
  13. Mask all the Petri dishes with a parafilm tape and place all the Petri dishes upside down into the incubator for four days at 30 to 40 degrees celsius
  14. Check and measure all the Petri dishes sterilised filter paper disk diameter of the inhibition of bacterial growth (the diameter of the bacteria killed) with the 15 cm ruler every day and enter the data into the google sheet. Afterwards, we drew out a histogram. 
  15. Take out the Petri dishes every four days to record the findings and take photos with a ruler and a label
  16. Repeat steps 1 - 16 if the experiment goes wrong
  17. Redo experiment after a successful experiment

Experiment B

  1. Collect sample bacteria from the canteen uncle.
  2. Set up 5 Petri dishes.
  3. Light up the alcohol lamp to create an up-draft to prevent dust from entering the Petri dishes. 
  4. Put 12.5 ml of agar jelly into each petri dish.
  5. Use a cotton bud swab to spread 50ul of sample bacteria evenly from school canteen into each of the Petri dishes. Only put on safety goggles at this point to prevent bacteria from getting into the eye.  
  6. Put five different concentrations of Clorox into their respective test tubes (8ml of water and 2ml of Clorox, 6ml of water and 4ml of Clorox, 4ml of water and 6ml of Clorox, 2ml of water and 8ml of Clorox, and undiluted Clorox)   
  7. Leave 17.45ml as bacteria need carbon dioxide and oxygen to live. 
  8. Use a tweezer and soak each sterilised filter paper disk into each solution.  
  9. Soak a total of 5 sterilised filter paper disks of different solutions. 
  10. Place one different soaked sterilised filter paper disk into each petri dish. 
  11. Do not place any sterilised filter paper disk for the last one as it is the control variable. 
  12. Sterilise the tweezer after placing a soaked sterilised filter paper disk by putting it above the alcohol lamp to prevent the different soaps from mixing. 
  13. Name all the different Petri dishes with a sticker to a label. 
  14. Mask all the Petri dishes with a parafilm tape and place all the Petri dishes upside down into the incubator for four days at 30 to 40 degrees celsius
  15. Check and measure all the Petri dishes sterilised filter paper disk diameter of the inhibition of bacterial growth (the diameter of the bacteria killed) with the 15 cm ruler every day and enter the data into the google sheet. Afterwards, we draw out a histogram. 
  16. Take out the Petri dishes every four days to record the findings and take photos with a ruler and a label.
3.4 Data Analysis: Describe the procedures you will use to analyze the data / results. 
  1. Tabulate the data and determine the diameter of the inhibition of bacteria using a 15cm ruler
  2. Plot a histogram of the diameter of inhibition zone to the type of detergent




Conclusion: From the histogram, we can find out Dettol is the most effective followed by Mr Muscle, Clorox, Jif then Mama lemon. 
4. Risk Assessment and Management: Identify any potential risks and safety precautions to be taken.
Table 1: Risk Assessment and Management table 

RiskAssessmentManagement
Petri dish may break while handling with it and may hurt the studentsMediumCovering the petri dish with parafilm to prevent the petri dish falling apart and handling it with care will prevent any accidents.
If bacteria are handled wrong, we may get contaminated. Bacterias can cause illness like flu and stomach ache.HighWash hands after every experiment and use safety goggles and gloves.
Students might accidentally consume the bacteria or agar jelly.HighDo not bring food or drinks into the lab in order to not consume the bacteria and agar jelly as you thought it was the drinking water.
Some students might knock down the small bottles that have the soaps in it and leak into other chemicals that might cause devastating effects.HighPut the small bottles away from other chemicals.
The flames from the alcohol lamp may get near the flammable ethanol or any flammable chemicals.HighPut the flammable chemicals away from the alcohol lamp.
5. References: List at least three (3) major sources (e.g. science journal articles, books, internet sites) from your literature review. Choose the APA format and use it consistently to reference the literature used in the research plan.
List your entries in alphabetical order for each type of source.
Book (1)
Tom Besty, DC and Jim Keogh, RN. (2005). Microbiology Demystified.  United Kingdom: McGraw Hill
Journal Article (1)
Goodyear, N., Brouillette, N., Tenaglia, K.,  Gore, R. and Marshall, J. (2015). The effectiveness of three home products in cleaning and disinfection of Staphylococcus aureus and Escherichia coli on home environmental surfaces. Journal of Applied Microbiology, Volume 119, Issue 5, page 1207-1454
Winter, J. (2008). Bleach Activates a Redox-Regulated Chaperone by Oxidative Protein Unfolding. Cell, Volume 135, Issue 4, page 691-701.
Website (13)
Healthline, (2020). “Giardiasis”. Healthline. Retrieved 20 January from


Lenntech, (2020). “Disinfectants Sodium Hypochlorite”. Lenntech. Retrieved 5 February from 
Lexico, (2020). “Cleanliness”. Oxford. Retrieved 21 January from
https://www.lexico.com/en/definition/cleanliness
Lexico, (2020). “Bacterium”. Oxford. Retrieved 21 January from 
Mayo Clinic, (2017). “Toxoplasmosis”. Mayo Clinic. Retrieved 20 January from 
Melissa, S. (2009). “How Bleach Works”. Howstuffworks. Retrieved 5 February from
Meredith, J. (2019). "How to grow bacteria in a Petri dish." Wikihow. Retrieved 14 January 2020 from
Solvhealth, (2020). “Cold”. Solvhealth. Retrieved 20 January from
https://www.solvhealth.com/health-a-z/cold
Solvhealth, (2020). “Good Hygiene”. Solvhealth. Retrieved 20 January from 
Solvhealth, (2020). “Flu”. Solvhealth. Retrieved 20 January from
https://www.solvhealth.com/health-a-z/flu
Unilever Australasia (2020). “Jif.” Unilever. Retrieved January 20, 2020 from 
U.S Food & Drug administration, (2020). “5 things to know about Triclosan”. U.S Food & Drug administration. Retrieved 20 January from
https://www.fda.gov/consumers/consumer-updates/5-things-know-about-triclosan
Wayne C, (2015). “ISS S205 Group A”. ISS S205 Group A. Retrieved 20 January from 
Web MD, (2020). “Gastroenteritis ("Stomach Flu")”. Web MD. Retrieved 20 January from https://www.webmd.com/digestive-disorders/gastroenteritis#1
World Health Organisation, (2019). “Sanitation”. World Health Organisation. Retrieved 17 January 2020 from https://www.who.int/news-room/fact-sheets/detail/sanitation
6. Bibliography: List at least three (13) major sources (e.g. science journal articles, books, internet sites) from your literature review. Choose
the APA format and use it consistently to reference the literature used in the research plan. List your entries in alphabetical order for each type of source. It should also include at least the 3 entries in the References section.
Book (1)
Tom Besty, DC and Jim Keogh, RN. (2005). Microbiology Demystified.  United Kingdom: McGraw Hill
Journal Article (1)
Goodyear, N., Brouillette, N., Tenaglia, K.,  Gore, R. and Marshall, J. (2015). The effectiveness of three home products in cleaning and disinfection
of Staphylococcus aureus and Escherichia coli on home environmental surfaces. Journal of Applied Microbiology, Volume 119, Issue 5, page 1207-1454
Winter, J. (2008). Bleach Activates a Redox-Regulated Chaperone by Oxidative Protein Unfolding. Cell, Volume 135, Issue 4, page 691-701.
Website (13)
Healthline, (2020). “Giardiasis”. Healthline. Retrieved 20 January from


Lenntech, (2020). “Disinfectants Sodium Hypochlorite”. Lenntech. Retrieved 5 February from 
Lexico, (2020). “Cleanliness”. Oxford. Retrieved 21 January from
https://www.lexico.com/en/definition/cleanliness
Lexico, (2020). “Bacterium”. Oxford. Retrieved 21 January from 
Mayo Clinic, (2017). “Toxoplasmosis”. Mayo Clinic. Retrieved 20 January from 
Melissa, S. (2009). “How Bleach Works”. Howstuffworks. Retrieved 5 February from
Meredith, J. (2019). "How to grow bacteria in a Petri dish." Wikihow. Retrieved 14 January 2020 from
Solvhealth, (2020). “Cold”. Solvhealth. Retrieved 20 January from
https://www.solvhealth.com/health-a-z/cold
Solvhealth, (2020). “Good Hygiene”. Solvhealth. Retrieved 20 January from 
Solvhealth, (2020). “Flu”. Solvhealth. Retrieved 20 January from
https://www.solvhealth.com/health-a-z/flu
Unilever Australasia (2020). “Jif.” Unilever. Retrieved January 20, 2020 from 
U.S Food & Drug administration, (2020). “5 things to know about Triclosan”. U.S Food & Drug administration. Retrieved 20 January from https://www.fda.gov/consumers/consumer-updates/5-things-know-about-triclosan
Wayne C, (2015). “ISS S205 Group A”. ISS S205 Group A. Retrieved 20 January from 
Web MD, (2020). “Gastroenteritis ("Stomach Flu")”. Web MD. Retrieved 20 January from https://www.webmd.com/digestive-disorders/gastroenteritis#1
World Health Organisation, (2019). “Sanitation”. World Health Organisation. Retrieved 17 January 2020 from
https://www.who.int/news-room/fact-sheets/detail/sanitation

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