Rationale

Antibiotics are vital in reducing infectious diseases in humans, livestock, and animals
Developing Homeopathic Solutions to Minimize Antibiotic Procedure (Summarized)
worldwide. However, numerous infections including tuberculosis, salmonellosis,
pneumonia, and gonorrhea are becoming harder to treat as antibiotic resistant
Resistance to Tetracycline Through the tetB and tetM Genes Concentration gradient to test for homeopathy

Along With 3 Other Commonly Used Antibiotics

mutations in the microorganisms cause the antibiotics to become less effective.
Therefore, causing the antibiotics to lose their ability to kill harmful bacteria and
prevent infection. The rise of antibiotic resistance is considered one of the greatest
public health issues according to the World Health Organization, and the CDC reports
that antibiotic resistance may be the cause of 23,000 deaths world wide . Just as
(A 2nd Year Study) eDNA Extraction from sod plugs
bacteria in our bodies can become resistant to antibiotics, the same can happen with By: Josie Byers
animals, spreading infections from animals into consumer’s bodies and increasing 11th Grade
chances of infection. Essential oils and herbal remedies show promise to be a possible
remedy for the spread of antibiotic resistance due to their antimicrobial activity and may
Polymerase chain reaction to amplify eDNA
have the ability to reverse the beta-lactam antibiotic resistance.
Concentration Gradient Data
Problem Gel electrophoresis to test for tetB and tetM genes
Find or create a homeopathic solution that doesn’t disrupt soil health and can Data Concentrations Data Summary and Analysis Solutions Tested
reverse the presence of an antibiotic resistant gene.
Day Total
Key: 1. Lavender tea
Hypotheses
Treatment Day 1 2 Day 3 Day 4 change
Zone of inhibition to test for resistance to 3 other antibiotics
Lavender 1A 1 1 1 1 0
1 = 75% *100% already Treatment Average Change
2. Cinnamon oil
3. Cinnamon & Lavender oil
Lavender 1B 1 1 1 1 0
1. All treatment groups will rid the eDNA of the tetB and tetM genes coding deemed to not Lavender 0

for resistance to tetracycline

Lavender 2A 1 1 1 1 0
2 = 50% be
Lavender 2B 0 0 0 0 0
4. Control
Previously Determined
Cinnamon 3.5
2. All treatment groups will increase the zone of inhibition for penicillin, Lavender 3A 2 2 2 2 0 3 = 25% homeopathic*
ampicillin, and streptomycin Lavender 3B 1 1 1 1 0
4 = 10%
Mix 4.875 5. Lavender oil (Previously)
2021-2022: Determined large animal farming operations (LAFOS) contributed to the
3. The mix will perform the best at increasing riding the eDNA of resistance
Lavender 4A 1 1 1 1 0
Control 0

genes and increasing the zone of inhibition for penicillin, ampicillin, and
Lavender 4B 0 0 0 0 0
presence of antibiotic resistance and that the mutation is very prevalent near these
Cinnamon 1A 3 4 7 8 5
areas.
streptomycin
4. 100% concentration of oil will be a safe homeopathic solution to be sprayed
Cinnamon 1B

Cinnamon 2A
2

2
4

2
5

4
7

5
5

3
Most promising solution: Cinnamon essential oil at 10% concentration
on plants Cinnamon 2B 4 4 6 8 4

Cinnamon 3A 0 0 0 3 3

Zone of Inhibition Data Analysis

Null Hypotheses
Cinnamon 3B 0 0 2 4 4

Cinnamon 4A 1 1 1 2 1
The zone of inhibition test was concluded to test for antibiotic resistance to penicillin,
1. The treatments will not rid the eDNA of the tetB or tetM genes if originally ampicillin, and streptomycin. Similarly, eDNA extraction, a polymerase chain reaction,
Cinnamon 4B 1 1 1 4 3

Mix 1A 0 2 3 4 4 Testing: Antibiotic resistance to penicillin, ampicillin, and streptomycin

present. Mix 1B 2 3 4 4 2 and a gel electrophoresis were used to determine antibiotic resistance to tetracycline.
2. There will be no change in the zone of inhibition for penicillin, ampicillin, Mix 2A 1 2 4 4 3
Greater zone of inhibition = less antibiotic resistance The hypothesis that all treatment groups would increase the zone of inhibition was
Mix 2B 2 4 7 7 5
and streptomycin before and after treatment Mix 3A 0 1 5 6 6 partly supported. For one, the average penicillin zone of inhibition changed from 0 cm
3. The treatments will all be equally as effective at reversing the tetB and Mix 3B 2 3 4 7 5 for the control group to 1.45 cm for the mixed group. After conducting a two-way
Averages
tetM genes ANOVA test between the control and mix group for streptomycin, a p-value of
Mix 4A 0 1 7 8 8
Identifier Penicillin Ampicillin Streptomycin Penicillin Ampicillin Streptomycin
Mix 4B 1 2 5 7 6
4. The treatments will all be equally as effective at increasing zone of <0.01(below 0.05 alpha value) led to the rejection of a null hypothesis that there
Control A 0 0 1.4 No
Control 1A 2 2 2 2 0 Control B 0 0.8 1.3 Yes No
Penicillin Ampicillin Streptomycin

inhibition for penicillin, ampicillin, and streptomycin Control 1B 1 1 1 1 0 Control 2A 0 0.7 2.7 No No
would be no change in antibiotic resistance between the treatment and control groups.
Control 2A 4 4 4 4 0 Control 0 0.6 2.167
Similarly, between the cinnamon and control groups, there was statistical significance
Control 2B 0 0 2.9 No

Control 2B 2 2 2 2 0 Control 3A 0 1.1 2.4 No No

for both the ampicillin and penicillin groups (0.02 and 0.04, respectively).
Engineering Goals
Lavender 0.583 1.02 2.15 Control 3B 0 1 2.3 No No
Control 3A 0 0 0 0 0
Lavender 1A 1.2 1.3 1.9 Yes No No

Furthermore, the bacteria present in the zone of inhibition before and after treatment
Control 3B 2 2 2 2 0 Lavender 1B 0.6 0.8 1.4 Yes No No
Cinnamon 0.733 1.317 2.2
1. Determine if the engineered solution is more effective at reversing the Control 4A 2 2 2 2 0 Lavender 2A 1.1 2.2 Yes No

presence of the tetB and tetM genes Control 4B 1 1 1 1 0

edx Mix 1.45 1.367 2.7
Lavender 2B
Lavender 3A
0
0.6
1.8
1.2
2.4
2.5
Yes
Yes
Yes
No
No
No
were measured. The control group had 49.3% complete antibiotic resistance (no zone
2. Determine if the engineered solution is more effective at increasing zone of Lavender 3B 0 0 2.5 No of inhibition), 34.8% partial antibiotic resistance (zone of inhibition with some bacteria
Cinnamon 1A 0.6 1.4 2.7 Yes No Yes

inhibition for ampicillin, penicillin, and streptomycin Gel Electrophoresis Data Cinnamon 1B
Cinnamon 2A
0
0.9
1.4
1.3
2.8
1.9
Yes
Yes
Yes
Yes
No
Yes
present), and 15.9% no antibiotic resistance (no bacteria in the zone of inhibition). In
3. Create a solution at the right concentration that will not harm the plants Cinnamon 2B
Cinnamon 3A
0.6
1.2
1.3
1.3
1.5
2
Yes
Yes
Yes
Yes
No
No
comparison, the mixed group had 41.7% partial antibiotic resistance and 58.3% no
Treatment tetB tetM DNA extracted
Cinnamon 3B
Mix 1A
1.1
0
1.2
0
2.3
2.4
Yes
Yes
No
Yes
No
No
antibiotic resistance. They further indicated a decrease in antibiotic-resistant bacteria in
Lavender Yes Yes Yes
Mix 1B 0 1.1 2.2 Yes No No
the treatment groups. Finally, cinnamon and the mix reversed the antibiotic gene for
Future Research
Lavender Yes No Yes Mix 2A 1.4 1.7 3.5 Yes No No

tetracycline in all of the PCR’s conducted but only sometimes in the lavender group.
Lavender No Yes Yes Mix 2B 1.5 1.3 2.7 Yes No No
Cinnamon No No Yes Mix 3A NA NA NA

1. Evaluate how effective the treatments are at reversing different antibiotics Cinnamon
Cinnamon
No
No
No
No
Yes
Yes
Mix 3B NA NA NA Therefore, partially supporting the hypothesis that the treatment groups would
including aminoglycosides or cephalosporins Mix No Yes Yes
Control 4A

Control 4A
0

0
0

1.3
1.3

1.7 Yes
Yes

Yes
decrease the presence of the tetB and tetM genes.
Mix No Yes Yes
2. Compare the efficiency of lavender and cinnamon to other possible solutions Mix No No Yes Control 5A 0 0 1 No

Mix No Yes Yes

including garlic and lemon
Control 5A 0 0 1.2 Yes
Mix No Yes Yes
Control 6A 0 0 0.9 Yes

3. Test different concentrations and compare levels of homeopathy with

Mix No Yes Yes

Conclusion
Control 6A 0 0.4 1.4 Yes Yes
Control Yes Yes Yes

effectiveness in reversing antibiotic resistance

Control 4B 0 1.2 2 Yes Yes
Control Yes Yes Yes

More and more bacteria are growing resistant to antibiotics meant to kill them.
Control Yes Yes Yes Control 4B 0 1 1.9 Yes Yes

4. Engineer more complicated mixes of homeopathic agents and compare Positive Control Yes Yes Yes Control 5B 0 1.1 1.4 Yes Yes
Positive Control Yes Yes Yes Control 5B 0 1 1.4 Yes No
These bacteria can pick up antibiotic-resistant genes through horizontal or vertical gene
efficiency Positive Control Yes Yes Yes
Control 6B 0 0 1.3 Yes
Positive Control
Positive Control
Yes
Yes
Yes
Yes
Yes
Yes
Control 6B 0 1.4 1.5 Yes Yes transfer, and through evolution by natural selection, these genes have become more
Positive Control Yes Yes Yes Control 4C 0 0 1.2 No
abundant. Bacteria not being killed by antibiotics leads to increasing the number of
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Control 4C 0 0 1.4 No
Negative Control No No No
Control 5C 0 0 1.4 Yes infections, higher hospital bills, and increased risks of death or disability. To solve this
issue, the experiment attempted to find homeopathic solutions that could decrease the
Negative Control No No No Control 5C 0 0.5 1.5 Yes Yes
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Negative Control No No No
TetB and TetM Genes Control 6C

Control 6C
0

0
0.4

1.1
1.7

1.6
Yes

Yes
No

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66% Yes 33% 66% Yes 33%
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Mix 100% No
83% Yes 17%
No 100% Yes
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