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The Efficacy of Natural Antimicrobials

The Efficacy of Natural Antimicrobials. Velma C. Claypool Undergraduate Student Cellular and Molecular Concentration Department of Biology Tennessee Technological University Cookeville, TN 38505. Introduction. Serafino et al (2008): little scientific research

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The Efficacy of Natural Antimicrobials

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  1. The Efficacy of Natural Antimicrobials Velma C. Claypool Undergraduate Student Cellular and Molecular Concentration Department of Biology Tennessee Technological University Cookeville, TN 38505

  2. Introduction Serafino et al (2008): little scientific research Potential healing properties beyond herbal practitioners (Nayak & Pereira 2006) Multiple agents produce ideal treatment approach (Misner 2007) Drug resistance necessitates new therapeutic approaches (Mondello et al 2006) Tea tree oil: antiseptic, anti-inflammatory properties (Carson et al 2006) 21 essential oils : 6 bacterial species (Prabuseenivasan et al 2006)

  3. Objective: Measure natural antimicrobial agent efficacy Hypothesis: Natural substances reduce bacteria percentage Null Hypothesis: No bacterial reduction with natural antimicrobial

  4. Methods and Materials Experimental Design (link) Data Form (link) Diluted bacterial suspension: 108 colonies / mL (1,000,000,000 per mL) 1 million bacteria to each slide Agents tested for antimicrobial efficacy (Fig. 1) Carried out as shown in figures 2-5

  5. Methods and Materials • Agents tested for antimicrobial efficacy (Fig.1) Figure 1: (L to R) Neosporin ointment, Honey (Mellifora), Lemon oil (Citrus limonum), Tea tree oil (Melaleuca alternifolia), Neem oil (Azadirachta indica)

  6. Methods and Materials Figure 2: Embedding slides with diluted bacterial suspension.

  7. Methods and Materials Figure 3: Performing wipe test on dried bacterial slides with antimicrobial agents. Wipe Test (Rutala, W., and D. Weber 2001)

  8. Methods and Materials Figure 4: Streaking plates with swab from treated bacterial slides.

  9. Methods and Materials Figure 5: Streaked plates placed inside incubator overnight to determine bacterial growth.

  10. Methods and Materials Statistical method used (Kirkman 1996) The results of an unpaired t-test Mean value comparison (E:S) 7.42 : 62.6 t= -1.75 sdev= 54.7 (standard deviation)

  11. Methods and Materials Group A: Agents vs. E. coli Number of items= 6 (0.00 0.00 0.00 0.00 1.50 43.0) Mean = 7.42 95% confidence interval for Mean: -42.36 thru 57.19 Standard Deviation = 17.4 Hi = 43.0 Low = 0.00 Median = 0.00 Average Absolute Deviation from Median = 7.42

  12. Methods and Materials Group B: Agents vs. Staph. aureus Number of items= 6 (6.00 8.00 10.5 63.0 88.0 200) Mean = 62.6 95% confidence interval for Mean: 12.81 thru 112.4 Standard Deviation = 75.4 Hi = 200. Low = 6.00 Median = 36.8 Average Absolute Deviation from Median = 54.4

  13. Methods and Materials • T-test only including natural agents • High bacterial count of water removed • t= -1.71 • sdev= 18.9

  14. Methods and Materials • Group A: Agents vs. E. coli • Number of items= 4 (0.00 0.00 0.00 0.00) • Mean = 0.00 95% confidence interval for Mean: -23.16 thru 23.16 • Standard Deviation = 0.00 Hi = 0.00 Low = 0.00

  15. Methods and Materials • Group B: Agents vs. Staph. aureus • Number of items= 4 (8.00 10.0 10.5 63.0) • Mean = 22.9 95% confidence interval for Mean: -0.2876 thru 46.04 • Standard Deviation = 26.8 Hi = 63.0 Low = 8.00 • Median = 10.2 Average Absolute Deviation from Median = 13.9

  16. Results Distinct results observed between bacterial strains Disk diffusion proposed interesting results (Figs 6,7) Plates containing slides produced unreliable results Average colonies present taken per agent (Table 1)

  17. Results Figure 6: Staph. aureusdisk diffusion left to right (top): honey, water, Neosporin and (bottom): lemon oil, tea tree oil, neem oil.

  18. Results Figure 7: E. coli disk diffusion left to right (top): lemon oil, tea tree oil, neem oil and (bottom): honey, water, Neosporin.

  19. Results Table 1: Bacterial colonies present after wipe test (average per two slides)

  20. Results Figure 8: Bacterial colonies observed on E. coli control plates. E. coli controls (Figure 8)

  21. Results Figure 9: Bacterial colonies observed on Staph. aureus plates. Staph. aureus controls (Figure 9)

  22. Discussion • Results agree Serafino et al (2008) • Tea Tree oil possess antimicrobial properties • Further research required - biologically active compounds • Alternative treatments more acceptable • Plant extract use beyond herbal medicine • Results agree Nayak & Pereira (2006): therapeutic efficacy • As per Misner’s (2007) findings: • Essential oil compounds inhibit bacterial growth • Bacterial count remarkably reduced, esp. E. coli

  23. Discussion • Differences with Mondello et al (2006): tea tree • Specific bioactive constituent not tested • Agree with strong antimicrobial properties shown • Interest in natural medicinals parallels increased awareness side-effects conventional drugs • Results agree Prabuseenivasan et al (2006) • Drug resistant pathogens threaten conventional treatments • Rich source biologically active compounds • Further research = potential treatments (diseases & pathologies) • Results also agree Shaneyfelt et al (2006) • Research substantiates use • Natural immune mediators enhance innate response

  24. Conclusion Natural substances posses varying antimicrobial properties. Differences observed between control, treatment groups. Natural substances stronger: gram-negative bacteria Reject null hypothesis: no bacterial reduction Accept alternative hypothesis: natural antimicrobial reduces bacterial colonies Further research provides efficacy – alternative treatments

  25. References Carson, C., K. Hammer, T. Riley. 2006. Melaleuca alternifolia (Tea Tree) oil: a review of antimicrobial and other medicinal properties. Clinical Microbiological Review 19:50-62. Kirkman, T. 1996. Statistics to Use. http://www.physics.csbsju.edu/stats/ (December 3, 2008). Misner, B. 2007. A novel aromatic oil compound inhibits microbial overgrowth on feet: a case study. Journal of the International Society of Sports Nutrition 4:3.

  26. References Mondello, F., F. De Bernardis, A. Girolamo, A. Cassone, G. Salvatore. 2006. In vivo activity of terpinen-4-ol, the main bioactive component of Melaleuca alternifolia Cheel (tea tree) oil against azole-susceptible and –resistant human pathogenic Candida species. BMC Infectious Diseases 6:158. Nayak, B., and Pereira, L. 2006. Catharanthus roseus flower extract has wound-healing activity in Sprague Dawley rats. BMC Complementary and Alternative Medicine 6:41. Prabuseenivasan, S., M. Jayakumar, and S. Ignacimuthu. 2006. In vitro antibacterial activity of some plant essential oils. BMC Complementary and Alternative Medicine 6:39. Rutala, W., and D. Weber. 2001. New disinfection and sterilization methods. Emerging Infectious Diseases 7.

  27. References Serafino, A., P. Vallebona, F. Andreola, M. Zonfrillo, L. Mercuri, M. Federici, G. Rosi, E. Garaci, P. Pierimarchi. 2008. Stimulatory effect of Eucalyptus essential oil on innate cell-mediated immune response. BMC Immunology 9:17. Shaneyfelt, M., A. Burke, J. Graff, M. Jutila, M. Hardy. 2006. Natural products that reduce rotavirus infectivity identified by a cell-based moderate-throughput screening assay. Virology Journal 3:68.

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