Team 3

Spices of Life: The Antimicrobial Properties of Cinnamon, Clove, and Garlic on Streptococcus mutans

Lola Agabalogun, Preeti Gondi, Gary Hoffman, Tasneem Maner, Mehr Mathew, Xuewei Ouyang, Jason Roth, Ali Siddiqui, Rich Tamirian, Chelsea Taylor, Rachel Ying Spices

• Vegetable or plant substance that contributes flavor • Antimicrobial properties • Phytochemicals: benzene, sulfur • Extract v. Essential oils • Cinnamon, Clove, and Garlic Cinnamon

• Cinnamomum zeylanicum • Cinnamomum trees • Eugenol and cinnamaldehyde • Antimicrobial properties Clove

• Syzygium aromaticum • Dried product of immature flower buds • Natural medicine, antioxidants • Eugenol • Antimicrobial properties Garlic

• Allium sativum • Allicin • Role in plant • Effects in humans • Antimicrobial properties

Allicin Synergy

• Increase in activity • Additive vs. antagonistic effects Streptococcus mutans

• Non-motile bacterium found in human mouth o causes tooth decay o found in crevices & pits of teeth • Uses sucrose to generate a sticky polysaccharide → the bacteria to stick to one another → lactic acid → plaque Mouthwash

Antiseptic solutions: • Fluoride - prevents dental caries • Hydrogen peroxide – whitens teeth • Chlorhexidine - strong antimicrobial Purpose

• To determine the antimicrobial effects of garlic, cinnamon, and clove on S. mutans in various concentrations • To test several mouthwashes to determine which is most effective at killing S. mutans in the mouth Hypothesis

• In order of decreasing effectiveness against bacteria: o Garlic aqueous extract o Cinnamon aqueous extract o Clove aqueous extract • Synergistic effects: garlic and cinnamon together would be most effective at killing S. mutans Materials/Methods

20g/100mL

Garlic

• Boiled- 15 minutes • Filtered Clove

• Boiled- 15 minutes • Centrifuged

Cinnamon

• Boiled- 15 minutes • Let stand 20 minutes • Filtered

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Minimum Inhibitory Concentration

0.00 μL 0.680 μL 1.25 μL 2.50 μL 5.00 μL 10.0 μL cinnamon garlic

clove control

Each well o 1.5 mL tryptic soy broth o 10 μL diluted spice extract o 10 μL S. mutans culture

Materials/Methods

Paper Disk Diffusion Method

10 μL garlic

60 μL 20 μL cinna clove

0 μL 0 μl

30+ 30+ 50 μL 30 μL 30 μl 30 μl

30+ 40 μL 30 μl Materials/Methods

Mouthwash Experiment • 10 μL on each filter disk • Oasis • ACT Bubble Gum Blowout • Scope Outlast • Listerine Antiseptic • Listerine Zero • Crest Pro-Health • CareOne Anti-Septic • Cepacol Antibacterial Rinse

Indication of Antimicrobial Activity

Zone of Inhibition Results Results Results Results Interpretation of Results

• Spices: No zones of inhibition → spices did not kill any bacteria

• Mouthwashes: Zones of inhibition → effective in killing bacteria Possible Reasons for Unsupported Hypothesis

• Ineffective extraction methods o vacuum filtration o ethanol & other compounds o too much boiling • Powdered state of the spices o heating during processing o dried already • Use of extract instead of essential oils

Unlikely Possibilities

• Paper disk diffusion method o worked for mouthwashes o circles of clove seen around some disks • Thickly spread bacteria o but thinly spread for second experiment • Not enough extract used o 60 μL used in second experiment o some also soaked Error Analysis

• Issues with sterilization o led to contamination • Overboiling and not stirring • Non-uniform methods of extraction • Spices transferred and concentration lost • Pipetting errors Inhibitory Properties of Manufactured Mouthwashes Explanation of Our Mouthwash Results

“helps “kills the prevent germs that plaque and cause bad gingivitis” breath” Suggestions for Future Scientists

Before the experiment • Consider testing various spices • Consider testing various herbs • Find appropriate method • Consider testing essential oils instead of extracts Suggestions for Future Scientists

During Preparation • Use fresh spices • Boil in ethanol or other solvent • Use vacuum filtration

Practical Applications

• Imitate home-use method o Use of common household items • Home-use method was ineffective • However, spice mouthwash is possible. o extracted in lab o sold commercially • Process/production → not completely natural o still attractive to consumers in comparison to chemical mouthwash Conclusion

• Spice mouthwash cannot be prepared at home. o Require advanced methods of extraction

• Spices extracted with water are not as effective as mouthwashes.

• Spices still have potential as mouthwash. Acknowledgments

• Sponsors: • NJGSS: o John and Laura Overdeck o Dr. Miyamoto o The Crimmins Family Charitable o Dr. Surace Foundation o Myrna Papier o Ina Zucchi Family Trust o Anna-Mae Dinio-Bloch o NJGSS Alumni and Parents 1984 – o Janet Quinn 2012 o Rachel Sandler o Novartis o Lalita Nekkanti o Independent College Fund of New o The Counselors  Jersey o Watson Pharmaceuticals o Bayer Healthcare o Roche

THE END