CEL70:Notebook/BL2104 - 2009 - Group Project

From OpenWetWare
Jump to: navigation, search

<!-- sibboleth --><div id="lncal1" style="border:0px;"><div style="display:none;" id="id">lncal1</div><div style="display:none;" id="dtext">02/28/2009</div><div style="display:none;" id="page">CEL70:Notebook/BL2104 - 2009 - Group Project</div><div style="display:none;" id="fmt">yyyy/MM/dd</div><div style="display:none;" id="css">OWWNB</div><div style="display:none;" id="month"></div><div style="display:none;" id="year"></div><div style="display:none;" id="readonly">Y</div></div>

Owwnotebook icon.png <sitesearch>title=Search this Project</sitesearch>

Antimicrobial effectiveness of Allium sativum preparation


The increased misuse of antibiotics has given rise to resistance in bacteria (Hawkey 2008), an evolutionary mechanism attributable to natural selection. Improved clinical procedures can decrease the occurrence of multiple drug-resistant microbes, as has been demonstrated with methicillin resistant staphylococcus aureus (MRSA) in American hospitals (Burton 2009), but since it cannot eliminate the problem alternative antimicrobial treatments have to be developed. It is apparently more difficult for bacteria to become resistant to natural biocides, therefore increasing the use of such is considered to be a sensible approach to preventing further resistant strains arising as well as treating those already established.


A natural biocide such as allicin (Cavallito 1944), has been demonstrated to exhibit significant antimicrobial properties (Holzgartner1992), a result mainly attributed to thiol-modifying and antioxidant abilities (Cavallito 1944, Wills 1956, Prasad 1998, Rabinkov 1998). Allicin is chemically unstable and reactive, it is hence not found in intact Allium sativum. Only damaging the compartments associates the non proteinogenic amino acid alliin and its enzyme allinase giving rise to the desired biologically active metabolite (Stoll1951).

Ever since the discovery of allicin’s antimicrobial effects, allicin or allicin derivatives have been marketed in health shops as ‘health promoters’. Due to allicin’s instability, the integrity of such preparations is questionable.


The objective of this experiment is to demonstrate the effectiveness of commercially available allicin preparations (which is found in garlic) on the skin bacteria Staphylococcus aureus. We expect a minor effect or no effect at all on bacterial growth due to allicin's chemical instability.


  • HEPES buffer
  • Staph. aureus culture
  • Distilled water
  • Glucose based agar
  • Glass spreader
  • Ethanol 70%
  • commercially available Allium sativum health product
  • Ph meter
  • HCl (aq) 1M
  • NaOH (aq) 1M


  1. HEPES buffer calibrated to ph 7.50 made up to 500ml
  2. Content of garlic capsules extracted.
  3. 5x 10x dilution series of Garlic
  4. 100ul Staph Aureus spread over series of Sucrose based agar plates using Aseptic technique
  5. 100ul of each sample spread over bacteria using Aseptic technique.
  6. One undiluted sample, 5x 10x dilution series.
  7. One control plate using 100ul HEPES buffer.
  8. Incubate at 37˚C for 24 hours
  9. Photograph and analyse results


All plates, regardless of the level of treatment, showed the same amount of bacterial growth. The experimental plates also displayed no difference in bacterial growth from the control plate.


The results show the antimicrobial ineffectiveness of the commercially available allium sativum preparation tested.


The commercially available allium sativum/allicin preparation tested demonstrated no antimicrobial effectiveness on S. aureus. This has important implications for the health product market, which commonly manufactures items based on new scientific trends. Clinical results may emphasize the benefits of these natural compounds but the evidence must be viewed critically, especially with regard to the conditions under which the results were obtained.


  1. Yamada Y and Azuma K. Evaluation of the in vitro antifungal activity of allicin. Antimicrob Agents Chemother. 1977 Apr;11(4):743-9. PubMed ID:856026 | HubMed [Azuma_1977]
  2. Bakri IM and Douglas CW. Inhibitory effect of garlic extract on oral bacteria. Arch Oral Biol. 2005 Jul;50(7):645-51. DOI:10.1016/j.archoralbio.2004.12.002 | PubMed ID:15892950 | HubMed [Bakri_2005]
  3. Burton DC, Edwards JR, Horan TC, Jernigan JA, and Fridkin SK. Methicillin-resistant Staphylococcus aureus central line-associated bloodstream infections in US intensive care units, 1997-2007. JAMA. 2009 Feb 18;301(7):727-36. DOI:10.1001/jama.2009.153 | PubMed ID:19224749 | HubMed [Burton_2009]
  4. Chester J. Cavallito, John Hays Bailey, "Allicin, the Antibacterial Principle of Allium sativum. I. Isolation, Physical Properties and Antibacterial Action", Journal of the American Chemical Society, 1944, volume 66, pp 1950 - 1951. Template:DOI [Cavallito_1944]
  5. Edris AE. Pharmaceutical and therapeutic potentials of essential oils and their individual volatile constituents: a review. Phytother Res. 2007 Apr;21(4):308-23. DOI:10.1002/ptr.2072 | PubMed ID:17199238 | HubMed [Edris_2007]
  6. Ellmore G. S. und Feldberg R. S. (1994): Allin Lyase Localization in Bundle Sheaths of the Garlic Clove (Allium sativum). In: Am. J. Bot, 81(1); 89–94 [Ellmore_1994]
  7. Principles and Practice of Disinfection, Preservation and Sterilization. Oxford: John Wiley & Sons, Ltd., 2007. ISBN:1405101997 [Fraise]
  8. Groppo FC, Ramacciato JC, Motta RH, Ferraresi PM, and Sartoratto A. Antimicrobial activity of garlic against oral streptococci. Int J Dent Hyg. 2007 May;5(2):109-15. DOI:10.1111/j.1601-5037.2007.00230.x | PubMed ID:17461963 | HubMed [Groppo_2007]
  9. Han J, Lawson L, Han G, and Han P. A spectrophotometric method for quantitative determination of allicin and total garlic thiosulfinates. Anal Biochem. 1995 Feb 10;225(1):157-60. PubMed ID:7778769 | HubMed [Han_1995]
  10. Hawkey PM. The growing burden of antimicrobial resistance. J Antimicrob Chemother. 2008 Sep;62 Suppl 1:i1-9. DOI:10.1093/jac/dkn241 | PubMed ID:18684701 | HubMed [Hawkey_2008]
  11. Holzgartner H, Schmidt U, and Kuhn U. Comparison of the efficacy and tolerance of a garlic preparation vs. bezafibrate. Arzneimittelforschung. 1992 Dec;42(12):1473-7. PubMed ID:1288512 | HubMed [Holzgartner_1992]
  12. Lawson, L.D. and Wang, Z.J., 1993. Pre-hepatic fate of the organosulfur compounds derived from garlic (Allium sativum). Planta Medica 59, pp. A688–689 [Lawson_1993]
  13. Lawson LD, Wang ZJ, and Hughes BG. Identification and HPLC quantitation of the sulfides and dialk(en)yl thiosulfinates in commercial garlic products. Planta Med. 1991 Aug;57(4):363-70. DOI:10.1055/s-2006-960119 | PubMed ID:1775579 | HubMed [Lawson_1991]
  14. Koch, H P. Biopharmaceutics of garlic's effective compounds. In: Koch H P, Lawson L D. , editors; Koch H P, Lawson L D. , editors. Garlic. The science and therapeutic application of Allium sativum L. and related species. Baltimore, Md: Williams & Wilkins; 1996. pp. 213–220. [Koch_1996]
  15. Mayeux PR, Agrawal KC, Tou JS, King BT, Lippton HL, Hyman AL, Kadowitz PJ, and McNamara DB. The pharmacological effects of allicin, a constituent of garlic oil. Agents Actions. 1988 Aug;25(1-2):182-90. PubMed ID:2847508 | HubMed [Mayeux_1988]
  16. Prasad K, Laxdal VA, Yu M, and Raney BL. Antioxidant activity of allicin, an active principle in garlic. Mol Cell Biochem. 1995 Jul 19;148(2):183-9. PubMed ID:8594422 | HubMed [Prasad_1966]
  17. Rabinkov A, Miron T, Konstantinovski L, Wilchek M, Mirelman D, and Weiner L. The mode of action of allicin: trapping of radicals and interaction with thiol containing proteins. Biochim Biophys Acta. 1998 Feb 2;1379(2):233-44. PubMed ID:9528659 | HubMed [Rabinkov_1998]
  18. Riddles PW, Blakeley RL, and Zerner B. Ellman's reagent: 5,5'-dithiobis(2-nitrobenzoic acid)--a reexamination. Anal Biochem. 1979 Apr 1;94(1):75-81. PubMed ID:37780 | HubMed [Riddles_1979]
  19. Sivam GP. Protection against Helicobacter pylori and other bacterial infections by garlic. J Nutr. 2001 Mar;131(3s):1106S-8S. PubMed ID:11238826 | HubMed [Sivam_2001]
  20. STOLL A and SEEBECK E. [The specificity of the alliinase from Allium sativum]. C R Hebd Seances Acad Sci. 1951 Apr 9;232(15):1441-2. PubMed ID:14831230 | HubMed [Stoll_1951]
  21. Talgoy MM, Bell AW, and Duckworth HW. The reactions of Escherichia coli citrate synthase with the sulfhydryl reagents 5,5'-dithiobis-(2-nitrobenzoic acid) and 4,4'-dithiodipyridine. Can J Biochem. 1979 Jun;57(6):822-33. PubMed ID:38891 | HubMed [Talgoy_1979]
  22. Tsao SM, Hsu CC, and Yin MC. Garlic extract and two diallyl sulphides inhibit methicillin-resistant Staphylococcus aureus infection in BALB/cA mice. J Antimicrob Chemother. 2003 Dec;52(6):974-80. DOI:10.1093/jac/dkg476 | PubMed ID:14585852 | HubMed [Tsao_2003]
  23. WILLS ED. Enzyme inhibition by allicin, the active principle of garlic. Biochem J. 1956 Jul;63(3):514-20. PubMed ID:13341914 | HubMed [Willis_1956]
All Medline abstracts: PubMed | HubMed