How Do Food Preservatives Affect the Growth of Microorganisms?
AbstractThis project uses liquid cultures and agar plates to investigate the effects of different concentrations of a food preservative on microbial growth.
The purpose of this project is to determine the effective concentration for anti-microbial food preservatives.
Andrew Olson, Ph.D., Science Buddies
- Youssefian, A., 2003. "How Do Additives Affect the Growth of Microorganisms?" California State Science Fair Abstract [accessed September 20, 2006] http://www.usc.edu/CSSF/History/2003/Projects/J1337.pdf.
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Last edit date: 2017-07-28
The problem of protecting food from spoilage has been with us since prehistoric times. The solutions to this problem have changed with advances in technology and knowledge about what causes food to spoil. This project will focus on retarding microbial growth, which is only one of the causes of food spoilage.
There are many ways that food can be spoiled. For example, oils in food can become oxidized, releasing free fatty acids that cause a bitter, rancid taste. Additionally, natural enzymes that take part in the ripening process of fruits and vegetables can remain active after harvest, causing spoilage. Different chemical preservatives have been developed to counteract each of these different mechanisms: "Preservatives can be categorized into three general types: antimicrobials that inhibit growth of bacteria, yeasts, or molds; antioxidants that slow air oxidation of fats and lipids, which leads to rancidity; and a third type that blocks the natural ripening and enzymatic processes that continue to occur in foodstuffs after harvest." (Dalton, 2002)
In order for an antimicrobial preservative to work, it must be used at the right concentration. Ideally, it will disrupt microbial growth while at the same time preserving most of the nutritional value of the food.
To do this project, you should first do background research on methods of food preservation. Then, select an antimicrobial preservation method to test for your experiment. As an example, this project will test one of the oldest preservation methods by adding different concentrations of salt.
Terms and Concepts
To do this project, you should do research that enables you to understand the following terms and concepts:
- conditions for bacterial growth,
- minimum, maximum, and optimum values for temperature, pH, osmolarity.
- food additives,
- food preservation.
- What is meant by "effective concentration" for a chemical compound?
- What are the effective concentrations for the additives you intend to test?
- This article provides a general overview for methods of food preservation:
Brain, M., date unknown. "How Food Preservation Works," HowStuffWorks.com [accessed September 20, 2006] http://home.howstuffworks.com/food-preservation.htm.
- These articles provide an introduction to the different types of food preservatives, and what each is used for:
- Dalton, L., 2002. "What's That Stuff? Food Preservatives: Antimicrobials, Antioxidants, and Metal Chelators Keep Food Fresh," Chemical & Engineering News, American Chemical Society 80 (45): 40, available online [accessed September 20, 2006] http://pubs.acs.org/cen/science/8045/8045sci2.html.
- Foulke, J.E., 1998. "A Fresh Look at Food Preservatives," FDA Consumer, U.S. Food and Drug Administration [accessed August 29, 2013] http://www.nettally.com/prusty/formj.htm.
- This webpage has background information on nutritional requirements for bacterial growth. It is from an online textbook of bacteriology, which can be an excellent source of further information on bacteria:
Todar, K., 2002. "Nutrition and Growth of Bacteria," Todar's Online Textbook of Bacteriology, Department of Bacteriology, University of Wisconsin, Madison [accessed September 20, 2006] http://textbookofbacteriology.net/nutgro.html.
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Materials and Equipment
To do this experiment you will need the following materials and equipment:
- 4 low-salt chicken boullion cubes
- 500 mL hot water
- 10 glass jars with lids
- anti-microbial preservative; for example, you could try one of the following:
- electronic balance, such as the Fast Weigh MS-500-BLK Digital Pocket Scale, 500 by 0.1 G, available from Amazon.com
- masking tape
- marking pen
- disposable gloves. Can be purchased at a local drug store or pharmacy, or through an online supplier like Carolina Biological Supply Company. If you are allergic to latex, use vinyl or polyethylene gloves.
- inoculating loop. Sterile, disposable inoculating loops are available from Carolina Biological Supply Company. Alternatively, although it is less preferable, sterile cotton-tipped applicators (e.g., Q-tips from an unopened box) may be used instead. Sterile cotton-tipped applicators are also available from Carolina Biological Supply Company. Note: If you are not using sterile, disposable inoculating loops or applicators, but are using a metal loop instead, you will also need a bunsen burner to flame the loop.
- 40 nutrient agar plates. These are available from Carolina Biological Supply Company.
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For health and safety reasons, science fairs regulate what kinds of biological materials can be used in science fair projects. You should check with your science fair's Scientific Review Committee before starting this experiment to make sure your science fair project complies with all local rules. Many science fairs follow Intel® International Science and Engineering Fair (ISEF) regulations. For more information, visit these Science Buddies pages: Projects Involving Potentially Hazardous Biological Agents and Scientific Review Committee. You can also visit the webpage ISEF Rules & Guidelines directly.
- Dissolve 2 chicken broth cubes in 500 mL of hot water.
- Divide the solution into 10 glass jars (50 mL/jar).
- Following the table, add preservative at 4 different concentrations, and add nothing for the control condition. Make two replicates of each condition 10 jars total). Be sure that the salt you add is fully dissolved.
Label Amount Broth (mL) Amount Salt (g) Control 50 0 #1 2.5% 48.75 1.25 #2 5% 47.5 2.5 #3 10% 45 5.0 #4 20% 40 10
- If you use a preservative other than salt, you should do background research to come up with an estimate of the effective concentration. Make the effective concentration your #3 test condition. Condition #'s 1, 2, and 4 should be 1/4, 1/2, and 2 times this concentration, respectively.
- Take samples on 1st, 3rd, 5th and 7th days, and streak onto agar plates. To streak the agar plates, put on disposable gloves and use the inoculating loop (or sterile cotton-tipped applicator) to perform the quadrant streaking method (Inoculation: How to Put the Bacteria You Desire on a Petri Dish). Note: If you are using disposable loops or applicators, you will not need to use a bunsen burner to flame them since they should already be sterile.
- Be sure to properly label all plates with the test solution and day number.
- Tape plates closed, incubate (inverted) overnight, and count bacterial colonies.
- Does the number of colonies decrease as concentration of the preservative increases?
Bacteria are all around us in our daily lives and the vast majority of them are not harmful. However, for maximum safety, all bacterial cultures should always be treated as potential hazards. This means that proper handling, cleanup, and disposal are necessary. Below are a few important safety reminders. You can also see the Microorganisms Safety Guide for more details. Additionally, many science fairs follow ISEF Rules & Guidelines, which have specific guidelines on how bacteria and other microorganisms should be handled and disposed of.
- Keep your nose and mouth away from tubes, pipettes, or other tools that come in contact with bacterial cultures, in order to avoid ingesting or inhaling any bacteria.
- Make sure to wash your hands thoroughly after handling bacteria.
- Proper Disposal of Bacterial Cultures
- Bacterial cultures, plates, and disposables that are used to manipulate the bacteria should be soaked in a 10% bleach solution (1 part bleach to 9 parts water) for 1–2 hours.
- Use caution when handling the bleach, as it can ruin your clothes if spilled, and any disinfectant can be harmful if splashed in your eyes.
- After bleach treatment is completed, these items can be placed in your normal household garbage.
- Cleaning Your Work Area
- At the end of your experiment, use a disinfectant, such as 70% ethanol, a 10% bleach solution, or a commercial antibacterial kitchen/bath cleaning solution, to thoroughly clean any surfaces you have used.
- Be aware of the possible hazards of disinfectants and use them carefully.
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If you like this project, you might enjoy exploring these related careers:
Food Scientist or TechnologistThere is a fraction of the world's population that doesn't have enough to eat or doesn't have access to food that is nutritionally rich. Food scientists or technologists work to find new sources of food that have the right nutrition levels and that are safe for human consumption. In fact, our nation's food supply depends on food scientists and technologists that test and develop foods that meet and exceed government food safety standards. If you are interested in combining biology, chemistry, and the knowledge that you are helping people, then a career as a food scientist or technologist could be a great choice for you! Read more
MicrobiologistMicroorganisms (bacteria, viruses, algae, and fungi) are the most common life-forms on Earth. They help us digest nutrients; make foods like yogurt, bread, and olives; and create antibiotics. Some microbes also cause diseases. Microbiologists study the growth, structure, development, and general characteristics of microorganisms to promote health, industry, and a basic understanding of cellular functions. Read more
Agricultural InspectorWho works to protect the public health from food-borne illnesses? Agricultural inspectors. Everyone needs to eat, and agricultural inspectors work to ensure the quality and safety of the food supply to determine if they are in compliance. They also inspect farms, businesses, and food-processing plants to determine if they are in compliance with government food regulations and laws. Read more
Food Science TechnicianGood taste, texture, quality, and safety are all very important in the food industry. Food science technicians test and catalog the physical and chemical properties of food to help ensure these aspects. Read more
- Compare refrigerated and/or frozen vs. room-temperature broth.
- Try additional preservatives. What are their effective concentrations?
- Try preservatives with solid food, e.g., salt meat or pickled vegetables. See the Science Buddies project, Minimizing Bacteria in the Thawing and Cooking of Meat for a method of pureeing solid food in order to assess bacterial content. Be sure to establish baseline bacterial content by pureeing samples before treatment with preservatives.
- Compare starting sterile vs. not. If you have a pressure cooker, here is an experiment you can try. Follow canning procedures for three jars. For the other three, use normal conditions for cooking and left-over food storage in three separate jars. Test by plating samples from successive jars from each condition at 1 day, 1 week, 2 weeks.
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I took the idea from this project and specified my investigation by choosing a specific food preservative, Ascorbic acid, and testing to see how it would affect the growth of bacteria on agar plates. This idea was very great as it did not take much time but it still earned me a bronze medal on my first science fair ever!
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