Annex A - Group Research Proposal

Group Project Proposal (Science)
SCHOOL OF SCIENCE AND TECHNOLOGY, SINGAPORE


INVESTIGATIVE SKILLS IN SCIENCE


Names: Alya Sufiyah, Eleanor Chua, Devika Prem
Class: S2-07


Group Reference: F


A.    Indicate the type of research that you are adopting:


[ ] 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 School of Science and Technology, Singapore  


  1. Type & Category


Type of research: (3)


Category  –  (18- Microbiology)
Sub-category –  (C- Bacterial Microbiology)


Application of project relevant to SST Community, Society or the World:
Salmonella is a foodborne disease mainly caused by eggs and poultry. The contaminated item looks and smells normal, thus, it is almost impossible to identify the contaminated food items. Our project focuses on eggs which are easily contaminated. Naturally, bacteria enters the tissues prior to the shell formation or from faeces contamination when bacteria enters the pores of the shell. Washing of eggs facilitate the entry of microorganisms when water enters the pores. Water left on shell surface may also enhance the survival of microorganisms on eggshells (Centre for Food Safety, 2008).


Salmonella cases have been on a rise. In Singapore, the number of cases have risen by about 30% to close to 2,000 in 2015. An increasing trend has been observed, with the number of cases in 2016 exceeding 779 that were reported between January and June 20, 2015. (Qi, 2016). From here, we can see that majority of the public are unaware of the causes of Salmonella and ways to prevent it. This is also further proven based on a survey we conducted.


Through our survey, we noticed that even though only 28.9% of the respondents wash their eggs -  60% of them wash their eggs for 10 seconds. However, more than 80% of the respondents do not think there is a possibility of bacteria growth while washing it. Moreover, those who wash their eggs prefer to let it dry naturally than wipe it. However, the biggest concern is the fact that less that 40% of the total respondents know what Salmonella is.


Not only in Singapore, the yearly report of the Ozfoodnet network reported 11, 992 cases of Salmonella in Australia in the year 2010.


This project can determine the effect of washing on bacterial growth on eggshells. If the number of bacteria on eggs vary based on the different timing of washing eggs and the method of drying, we hope to be able to raise awareness on this issue through our findings.


C.    Write down your research title:
Investigation of the effect of washing on the growth of bacteria on eggshells.


D.   (a) Aim / question being addressed
Our aim is to find out if washing egg shells increase bacterial growth on the eggshell.


(b) Independent variable
The independent variable is amount of washing.


(c) Dependent variable
The dependent variable is bacterial growth.


(d) Controlled variables
  1. The type of egg
  2. The type of liquid used to wash the eggs
  3. The environment we are washing the eggs in
  4. The amount of time the eggs are placed in the fridge
  5. The storage location of the eggs (In the fridge)
  6. The method of drying
  7. The amount of agar placed on the dish


      (e) Hypotheses
  • The longer we wash the egg under running water, the greater the number of bacteria growing on the eggshells.
  • Unwashed eggs will have a lesser amount of bacteria on the shells as compared to all the other eggs which were washed.


E.    Method – Description in detail of method or procedures (The following are important and key items that should be included when formulating ANY AND ALL research plans.)


(a) Equipment list:
  • Medium sized eggs (12x)
  • 12 egg tray with holes to place eggs (1x)
  • Tap Water
  • Agar Plate (8x)
  • Incubation chamber (1x)
  • Cotton swab (10x)  
  • Gloves (3x)
  • Para films (1 roll)
  • detergent to clean egg tray/fridge
  • Fridge



(b) Diagrams
Step 1 Step 2


Step 3 Step 4 Step 5


Step 6: Record!


(c) Procedures: Detail all procedures and experimental design to be used for data collection


  1. Wash, dry and store the egg according to the below table.


Time Washed
Drying Method
Storage Location
Sample Name  
Not washed
-
Fridge (24 hours)
1A
Not washed
-
Fridge (24 hours)
1B
10 seconds
Naturally
Fridge (24 hours)
2A
10 seconds
Naturally
Fridge (24 hours)
2B
20 seconds
Naturally
Fridge (24 hours)
3A
20 seconds
Naturally
Fridge (24 hours)
3B
50 seconds
Naturally
Fridge (24 hours)
4A
50 seconds
Naturally
Fridge (24 hours)
4B


  1. Using a cotton swab, swab the outer surface of each egg shell by rolling the cotton swab on the shell in all direction for 5 seconds.  (to ensure all sides of the cotton swab have bacteria)


  1. Swipe the surface of the agar with the cotton swab. Careful to not tear the agar.
  1. Seal the agar plate with the parafilm tightly. Remember to label every plate.


  1. Incubate the inverted agar plates for 2-3 days at 32°C.


  1. After 3 days, take out the bacteria and observe them carefully. (Record the observations in Table A below.)
    1. Count the number of bacteria colonies.
    2. Look at the shape of the bacteria colonies.
    3. Look at the characteristics of the bacteria colonies.
    4. Draw an inference or conclusion from the bacteria colonies.  
Sample Name
Amount of Bacteria present (*using scale)
Shape and characteristics of Bacteria (describe clearly)
1A
little bacteria [3]
a few tiny punctiform colonies
2A
a lot of bacteria [7]
many tiny punctiform colonies (appears smudged and chalk-like)
3A
a lot of bacteria [7]
many tiny punctiform colonies (appears smudged and chalk-like)
4A
a lot of bacteria [9]
many clear tiny circular colonies
1B
little bacteria [3]
a few tiny punctiform colonies
2B
a lot of bacteria [8]
many tiny punctiform colonies (appears smudged and chalk-like)
3B
a lot of bacteria [7]
clear tiny punctiform colonies
4B
some bacteria [5]
many clear tiny circular colonies

*a scale of 1-10, in which 1 is the least and 10 is the most.


(d) Risk, Assessment and Management: Identify any potential risks and safety precautions to be taken.


1. Risk Assessment and Management table
What is the Risk
Assessment
How to Resolve?
Spilled water (When washing the egg) may be a slip hazard and can cause us to be injured (to a certain extent).
Medium
Be careful to not splash the water while washing eggs and be sure to wipe any spills.
As the experiment involves using the agar plate which is a glassware, there is a risk of breakage and cutting of the hands.
High
Always wear gloves when using the glassware. In case of any breakage, the cut to the hands will be minimized. Remember to be careful with the glassware and do not use bare hands to pick up any shards of glass in case of breakage. Inform a teacher of the breakage and dispose the glass into the appropriate ‘sharp’ bin.
We might accidentally spill or drop the agar from the agar plate which may cause skin irritation.
Low
Handle the agar plate with care and make sure that none of our friends touch the spilled agar.
We might accidentally come into contact with the bacteria, which causes us to get sick.
Medium
Wear gloves at all times of the experiment and make sure that the egg shell does not come into contact with any bodily fluids. Rinse with water thoroughly if contacted.


(e) Data Analysis: Describe the procedures you will use to analyze the data/results that answer research questions or hypotheses


  1. Rank the agar colonies to determine the bacteria present. We have adopted a scale of 1 to 10. This is done based on observation. We will also describe the shape and characteristics of bacteria growth. After that, we will take a picture and keep records. (Table A)
  2. After that, we will also use Adobe Photoshop CC to determine the RGB level for each photograph. We selected the area of the petri dish to determine the RGB levels. The greater the total value of R and G, which is yellow, the more bacteria is there.
  3. We then plot 4 graphs. Two graphs (Set A and B) for bacterial count vs washing time and another two (Set A and B) graphs for GR value vs washing time. With this, we can compare the graphs to determine whether the washing time has an impact on the growth of bacteria.

F. Bibliography: List at least five (5) major sources (e.g. science journal articles, books, internet sites) from your literature review. If you plan to use vertebrate animals, one of these references must be an animal care reference. 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.


(a) E-books
Pietro (2009). Retrieved January 22, 2017, from https://www.cambridge.org/core/books/salmonella-infections/E9ECF3B4E5D52BBEC8C883C53539303E [Reference]
 Giannella, R. A. (1996, January 01). Salmonella. Retrieved January 22, 2017, from          https://www.ncbi.nlm.nih.gov/books/NBK8435/ [Reference]


(b) Websites
Arvin, Irfan, Meng Hou, Raphael. (2015). 2. Methods. Retrieved January 20, 2017, from http://sst2015-s208iss-d.blogspot.sg/p/2-methods.html
Centre for Food Safety(2008) - Bacteria on Eggs – Should Eggs be Washed? (2008, December 19). Retrieved January 10, 2017, from http://www.cfs.gov.hk/english/multimedia/multimedia_pub/multimedia_pub_fsf_29_01.html
Gentry, R. F., & Quarles, C. L. (1972). The Measurement of Bacterial Contamination on Egg Shells *. Retrieved January 14, 2017, from http://ps.oxfordjournals.org/content/51/3/930.abstract
May, D., & Kiermeier, A. (2014). Effect of Egg Washing and Correlation between Eggshell Characteristics and Egg Penetration by Various SalmonellaTyphimurium Strains. Retrieved January 10, 2017, from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3951326/
Qi, L. J. (2016). MOH monitoring rising number of salmonella cases. Retrieved January 10, 2017, from http://www.channelnewsasia.com/news/singapore/moh-monitoring-rising/2910822.html
Nuffield Foundation (2011). Incubating and viewing plates. Retrieved January 25, 2017, from http://www.nuffieldfoundation.org/practical-biology/incubating-and-viewing-plates
Caidoz. (2017). Grow Bacteria in a Petri Dish. Retrieved January 25, 2017, from http://www.wikihow.com/Grow-Bacteria-in-a-Petri-Dish


(c) Video
EphedrineTV (2011). How to use a Micropipette Retrieved from https://youtu.be/8Afh_0IAfrQ

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