Which antibacterial substance: Yogurt (Lactobacillus bulgaricus)

or Cranberry Juice (Vaccinium subg. Oxycoccus) inhibits the

largest amount of gram-negative bacterial growth and how does

the concentration of the substances affect the inhibitancy of

gram-negative bacterial growth

Biology Extended Essay

1,411 Words

[sarah alobaidan]

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 Contents

I. Cover page..........................................................................................................................1

II. Table of Contents................................................................................................................2

1.Investigation..................................................................................................................................2

1.1 Research Question.................................................................................................................3

1.2 Background Information........................................................................................................3

2. Methodology................................................................................................................................4

2.1 Investigation...........................................................................................................................4

2.2 Hypothesis..............................................................................................................................5

3. Materials and procedure...............................................................................................................6

3.1 Variables................................................................................................................................6

3.2 Materials required..................................................................................................................6

3.3 Procedure (Part A).................................................................................................................6

3.4 Ethical considerations............................................................................................................7

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1.Investigation

1.1 Research Question

Which antibacterial substance: Yogurt (Lactobacillus bulgaricus) or Cranberry Juice (Vaccinium

subg. Oxycoccus) inhibits the largest amount of gram-negative bacterial growth and how does

the concentration of the substances affect the inhibitancy of gram-negative bacterial growth?

1.2 Background Information

Diseases and illnesses are caused by a variety of factors such as viruses, bacteria, and other

microorganisms. These diseases affect a large segment of the population and result in increased

health issues. Acute sore throat, often known as pharyngitis, is one of these ailments. Acute

throat infections are one of the most prevalent infectious disorders treated by family

practitioners, accounting for 2% to 4% of all family practitioner's visits. Viruses cause 85% to

95% of throat infections in people and children under the age of five; viruses cause around 70%

of throat infections in children aged five to fifteen, with the remaining 30% due to bacterial

infections.1 Sore throats are normally treated with penicillin or amoxicillin (Amoxil) properties

that can effectively constrain the growth of gram-positive and gram-negative bacteria and cure

the infection2.

However, with the increase of the over usage of such drugs, bacterial strains have developed a

greater resistance towards the drugs. There is scientific evidence that the usage or misuse of

biocides can contribute to the increased occurrence of bacteria that are resistant not only to

biocides but also to antibiotics.3 Due to this, this poses a significant problem for the people who

are affected by acute sore throat. Therefore, there has been a new spike of scientific interest in
1
Worrall, G. (2011, July). Acute sore throat. Canadian family physician Medecin de famille canadien. Retrieved December 25, 2021, from
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3135445/#b2-0570791
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AA;, A.-J. (n.d.). In vitro antibacterial activity of ibuprofen and acetaminophen. Journal of global infectious diseases. Retrieved December 25,
2021, from https://pubmed.ncbi.nlm.nih.gov/20606962/

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finding different solutions that are not bacteria resistant and possess the ability to inhibit bacterial

growth.

Cranberry Juice (Vaccinium subg. Oxycoccus) is common in many households. Generally known

as a house-hold remedy for sore throats, it’s a inexpensive beverage that is believed to treat

several aliments. There is evidence of the antibacterial properties of cranberry juice (Vaccinium

subg. Oxycoccus) like its ability to treat sore throat4. However, cranberry juice (Vaccinium subg.

Oxycoccus) is not the only substance that has caught scientists' attention. Recently, Yogurt

(Lactobacillus bulgaricus) has also been in recent intrest due to its 5.

However, the extent of the bacterial growth prevention effects on antimicrobial substances has

not been fully verified. If these substances have significant antibacterial effects on cultured

bacterial. Further investigation should be conducted to know whether if they are suitable

alternative treatments for sore throat.

2. Methodology

2.1 Investigation

The aim of this investigation is to find out which antimicrobial substance; Yogurt (Lactobacillus

bulgaricus) or Cranberry Juice (Vaccinium subg. Oxycoccus) can inhibit the largest amount of

gram-negative bacterial growth, proving to be the best inhibitor of gram-negative bacterial

growth. The diluted concentrations of the two substances are simulated to be the same as how

3
Effects of biocides on antibiotic resistance. (n.d.). Retrieved December 25, 2021, from
https://ec.europa.eu/health/scientific_committees/opinions_layman/en/biocides-antibiotic-resistance/biocides-antibiotic-resistance-greenfacts-
level2.pdf
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Sethi, R., & Govila, V. (2011, January). Inhibitory effect of cranberry juice on the colonization of streptococci species: An in vitro study.
Journal of Indian Society of Periodontology. Retrieved December 25, 2021, from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3134047/
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Mohammadi-Gouraji, E., Soleimanian-Zad, S., & Ghiaci, M. (2018, September 24). Phycocyanin-enriched yogurt and its antibacterial and
physicochemical properties during 21 days of storage. LWT. Retrieved December 25, 2021, from
https://www.sciencedirect.com/science/article/abs/pii/S0023643818307898

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majority of people uses it. When the best inhibitor is determined, the concentrations of the

substance will be varied to find the optimum concentration for the substance to inhibit bacterial

growth

The approach taken in this investigation is to use the Kirby-Bauer test on self-cultured bacteria

taken from the mouth and throat area. The test utilizes the usage of agar plates and filter paper

discs to show the extent of the inhibitation of the bacteria growth. Filter paper disc with the

substance will be placed into agar plates with the bacteria and after 24 hours, the area of bacteria

prevention will appear on the disc. This region shows the area of zero bacterial growth and

displays the amount of inhibitation of the bacterial growth.

The area of bacterial growth inhibited will be calculated using the formula of ellipse to take in

the regions of inhibitation with two different radii. Once the best inhibitor is found; it will be

diluted into five different concentrations inorder to find the best concentration for minimal

bacterial growth.

This investigation will be split into part A and part B, where the original answer of the variables;

which substance inhibits the largest amount of gram-negative bacterial growth and the latter

answers the second variable, how does the concentration of the substances affect the inhibitancy

of gram-negative bacterial growth.

2.2 Hypothesis

As prior research has shown that cranberry juice (Vaccinium subg. Oxycoccus) inhibits a large

amount of bacterial growth, it is hypothesized that cranberry juice (Vaccinium subg. Oxycoccus)

will inhibit the largest amount of bacterial growth, and as the concentration of cranberry juice

(Vaccinium subg. Oxycoccus) increases, the amount of bacterial growth inhibited increases

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3. Materials and procedure

3.1 Variables

Independent variables:

The first independent variable is the several types of antibacterial substances, particulary

cranberry juice and yogurt. The Second independent variable is the concentration of the

substance that inhibits the largest amount of bacterial growth

Dependent variables:

The dependent variable is the amount of inhibited bacterial growth on the ager plates, which is

measure by the region of inhibitation

Controlled variables

Table 1 showing the controlled variables

Controlled Variables How it is controlled

The location of the bacteria that was swabbed Inside the mouth and throat that was swabbed
The type of bacteria used The same inoculated colony was used for all trials
The brand of Cranberry Juice used Only the “Ocean sprray” brand was used
The volume of sterile nutrient broth used
Volume of gram-negative bacteria used for each
petri dish
Volume of antibacterial substance used per petri
dish
The total duration that the bacteria was left to be Each was culture for 48 hours (about 2 days)
cultured

3.2 Materials required

3.3 Procedure (Part A)

I. Preperation of agar plates

a. A petri dish is filled with nutrient agar and covered with a lid

b. The petri dish is then coverewd and sealed with parafilm

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 c. This is then repeated for the other samples

II. Preparation of bacterial cultures

a. A sterile swab is used to swab the inside of the mouth and throat for bacteria

b. The swab is then moved over to an agar plate

c. The petri dish is then inculbated for 48 hours (about 2 days)

d. Once visible the petri dish is then refrigerated

III. Prevetion of bacteria

a. A visible colony is chosen from the agar plate

b. A sample of the bacterial colony is then collected

c. The bacteria is then smeared on a new agar plate

d. The agar plate is then incubated for 48 hours (about 2 days)

e. Once the bacterial colonies are visible, the agar plates are then refrigiated

IV. Gram staining of bacterial colony

V. Preparation of bacterial broth

a. Sterile nutrient broth is measured by a syringe

b. inoculating loop is used to put the bacteria that was cultured in III into the nutrient

broth

VI. Dilution of antibacterial substances

a. The antibacterial substances are then diluted

VII. Kirby-Bauer test

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3.4 Ethical considerations

Thoughout this investigation, no personal information about the person who will be swabbed will

be asked or made public throughout this investigation. In addition, the subject must fill out a

consent form before the experiment, which will include information about the trial. The

swabbing of the mouth will be done with sterile equipment. The bacteria that will be cultivated

will be continuously monitored to ensure that they do not spread with the addition of protective

clothes and gear will be worn whenever working on the experiment.