, Volume 169, Issue 4, pp 287–295

In Vitro Investigation of Antifungal Activity of Allicin Alone and in Combination with Azoles Against Candida Species

  • Alireza Khodavandi
  • Fahimeh Alizadeh
  • Farzad Aala
  • Zamberi Sekawi
  • Pei Pei Chong


Candidiasis is a term describing infections by yeasts from the genus Candida, and the type of infection encompassed by candidiasis ranges from superficial to systemic. Treatment of such infections often requires antifungals such as the azoles, but increased use of these drugs has led to selection of yeasts with increased resistance to these drugs. In this study, we used allicin, an allyl sulfur derivative of garlic, to demonstrate both its intrinsic antifungal activity and its synergy with the azoles, in the treatment of these yeasts in vitro. In this study, the MIC50 and MIC90 of allicin alone against six Candida spp. ranged from 0.05 to 25 μg/ml. However, when allicin was used in combination with fluconazole or ketoconazole, the MICs were decreased in some isolates. Our results demonstrated the existing synergistic effect between allicin and azoles in some of the Candida spp. such as C. albicans, C. glabrata and C. tropicalis, but synergy was not demonstrated in the majority of Candida spp. tested. Nonetheless, In vivo testing needs to be performed to support these findings.


Allicin Candida spp. Fluconazole Ketoconazole Antifungal activity 


  1. 1.
    Rangel-Frausto MS, Wilblin T, Blumberg HM, Saiman L, Patterson J, Rinaldi M, et al. National epidemiology of mycoses survey (NEMIS): variations in rates of bloodstream infections due to Candida species in seven surgical intensive care units and six neonatal intensive care units. Clin Infect Dis. 1999;29:253–8.CrossRefPubMedGoogle Scholar
  2. 2.
    Banerjee SN, Emori TG, Culver DH, Gaynes RP, Jarvis WR, Horan T. Secular trends in nosocomial primary blood-stream infections in the United States. National Nosocomial Infections Surveillance System. Am J Med. 1991;91(3B):S86–9.CrossRefGoogle Scholar
  3. 3.
    Edmond MB, Wallace SE, McClish DK, Pfaller MA, Jones RN, Wenzel RP. Nosocomial bloodstream infections in United States hospitals: a three-year analysis. Clin Infect Dis. 1999;29:239–44.CrossRefPubMedGoogle Scholar
  4. 4.
    Ng KP, Madasamy M, Saw TL, Baki A, He J, Soo-Hoo TS. Candida biotypes isolated from clinical specimens in Malaysia. Mycopathol. 1999;144:135–40.CrossRefGoogle Scholar
  5. 5.
    Odds FC, Brown AJP, Gow NAR. Antifungal agents: mechanisms of action. Trends Microbiol. 2003;11:272–9.CrossRefPubMedGoogle Scholar
  6. 6.
    Hemaiswaryaa S, Kruthiventib AK, Doblea M. Synergism between natural products and antibiotics against infectious diseases. Phytomedicine. 2008;15:639–52.CrossRefGoogle Scholar
  7. 7.
    Ankri S, Mirelman D. Antimicrobial properties of allicin from garlic. Microbes Infect. 1999;2:125–9.CrossRefGoogle Scholar
  8. 8.
    Cai Y, Wang R, Pei F, Liang BB. Antibacterial activity of allicin alone and in combination with β -lactams against Staphylococcus spp. and Pseudomonas aeruginosa. J Antibiot. 2007;60(5):335–8.CrossRefPubMedGoogle Scholar
  9. 9.
    Yamada Y, Azuma K. Evaluation of the in vitro antifungal activity of allicin. Antimicrob Agent Chemother. 1977;11:743–9.Google Scholar
  10. 10.
    Shen JK, Davis LE, Cai Y. Antifungal activity in human cerebrospinal fluid and plasma after intravenous administration of Allium sativum. Antimicrob Agents Chemother. 1990;34(4):651–3.PubMedGoogle Scholar
  11. 11.
    Davis SR, Perrie R, Apitz-Castro R. The in vitro susceptibility of Scedosporium prolificans to ajoene, allitridium and a raw extract of garlic (Allium sativum). J Antimicrob Chemother. 2003;51:593–7.CrossRefPubMedGoogle Scholar
  12. 12.
    Tsao SM, Yin MC. In vitro antimicrobial activity of four diallyl sulphides occurring naturally in garlic and Chinese leek oils. J Med Microbiol. 2001;50:646–9.PubMedGoogle Scholar
  13. 13.
    Yoshida S, Kasuga S, Hayashi N, Ushiroguchi T, Matsuura H, Shizutoshi N. Antifungal activity of ajoene derived from garlic. Appl Environ Microbiol. 1987;53(3):615–7.PubMedGoogle Scholar
  14. 14.
    Lemar KM, Turner MP, Lloyd D. Garlic (Allium sativum) as an anti-Candida agent: a comparison of the efficacy of fresh garlic and freeze-dried extracts. J Appl Microbiol. 2002;93:398–405.CrossRefPubMedGoogle Scholar
  15. 15.
    Low CF, Chong PP, Yong PVC, Lim CSY, Ahmad Z, Othman F. Inhibition of hyphae formation and SIR2 expression in Candida albicans treated with fresh Allium sativum (garlic) extract. J Appl Microbiol. 2008;105(6):2169–77.CrossRefPubMedGoogle Scholar
  16. 16.
    Davis SR. An overview of the antifungal properties of allicin and its breakdown products–the possibility of a safe and effective antifungal prophylactic. Mycoses. 2005;48:95–100.CrossRefPubMedGoogle Scholar
  17. 17.
    Hughes BG, Lawson DL. Antimicrobial effects of Allium sativum L. (garlic), Allium ampeloprasum (elephant garlic), and Allium cepa (onion), garlic compounds and commercial garlic supplement products. Phytother Res. 1991;5:154–8.CrossRefGoogle Scholar
  18. 18.
    Feldberg RS, Chang SC, Kotik AN, Nadler M, Neuwirth Z, Sundstrom DC, Thompson NH. In vitro mechanism of inhibition of bacterial growth by allicin. Antimicrob Agents Chemother. 1988; 1763–1768.Google Scholar
  19. 19.
    MaoMao A, Hui S, YongBing C, JunDong Z, Yun C, Rui W, et al. Allicin enhances the oxidative damage effect of amphotericin B against Candida albicans. Int J Antimicrob Agents. 2009;33:258–63.CrossRefGoogle Scholar
  20. 20.
    Miron T, Rabinkov A, Mirelman D, Wilchek M, Weiner L. The mode of action of allicin: its ready permeability through phospholipid membranes may contribute to its biological activity. Biochim Biophys Acta. 2000;1463:20–30.CrossRefPubMedGoogle Scholar
  21. 21.
    National Committee for Clinical Laboratory Standards (NCCLS). Reference method for broth dilution antifungal susceptibility testing of yeasts. Approved standard NCCLS document M27-A2. Wayne PA NCCLS. 2002; 22.Google Scholar
  22. 22.
    Sahm DF, Washington JA II. Antibacterial susceptibility tests: dilution methods. Manual of clinical microbiology. Washington DC: American Society for Microbiology; 1995.Google Scholar
  23. 23.
    George M, Eliopoulos GM, Robert C, Moellering JR. Antimicrobial combinations. Antibiotics in laboratory medicine. Baltimore: The Williams & Wilkins; 1996.Google Scholar
  24. 24.
    Lebouvier N, Pagniez F, Duflos M, Le Pape P, Na YM, Le Bauta G, et al. Synthesis and antifungal activities of new fluconazole analogues with azaheterocycle moiety. Bioorgan Med Chem Lett. 2007;17:3686–9.CrossRefGoogle Scholar
  25. 25.
    Pereira-Cencil T, Del Bel Cury AA, Crielaard W, Ten Cate JM. Development of Candida-associated denture stomatitis: new insights. J Appl Oral Sci. 2008;16(2):86–94.Google Scholar
  26. 26.
    He QQ, Li K, Cao YB, Dong HW, Zhao LH, Liu CM, et al. Design, synthesis and molecular docking studies of novel triazole antifungal compounds. Chinese Chem Lett. 2007;18:663–6.CrossRefGoogle Scholar
  27. 27.
    Al-Mohsen I, Hughes WT. Systemic antifungal therapy: past, present and future. Annal Saudi Med. 1998;18:28–38.Google Scholar
  28. 28.
    Cauffman CA, Lynch JP. Fungal pneumonia. Pulmonary/respiratory therapy secrets. In: Parsons PE, Heffner JE (eds). Philadelphia: Hauley & Belfus Inc. 2001; 173.Google Scholar
  29. 29.
    Rogers T, Galgiani JN. Activity of Fluconazole (UK 49, 858) and Ketoconazole against Candida albicans in vitro and in vivo. Antimicrob Agent Chemother. 1986;30:418–22.Google Scholar
  30. 30.
    Shadkchan Y, Shemesh E, Mirelman D, Miron T, Rabinkov A, Wilchek M, et al. Efficacy of allicin, the reactive molecule of garlic, in inhibiting Aspergillus spp. in vitro, and in a murine model of disseminated aspergillosis. J Antimicrob Chemother. 2004;53:832–6.CrossRefPubMedGoogle Scholar
  31. 31.
    Pyun MS, Shin S. Antifungal effects of the volatile oils from Allium plants against Trichophyton species and synergism of the oils with ketoconazole. Phytomedicine. 2006;13:394–400.CrossRefPubMedGoogle Scholar
  32. 32.
    Elnima EI, Ahmed SA, Mekkawi AG, Mossa JS. The antimicrobial activity of garlic and onion extracts. Pharmazie. 1983;38:747–8.PubMedGoogle Scholar
  33. 33.
    Ogita A, Hirooka K, Yamamoto Y, Tsutsui N, Fujita K, Taniguchi M, et al. Synergistic fungicidal activity of Cu2 + and allicin, an allyl sulfur compound from garlic, and its relation to the role of alkyl hydroperoxide reductase 1 as a cell surface defense in Saccharomyces cerevisiae. Toxicology. 2005;215:205–13.CrossRefPubMedGoogle Scholar
  34. 34.
    Ogita A, Fujita K, Taniguchi M, Tanaka T. Dependence of synergistic fungicidal activity of Cu2 + and allicin, an allyl sulfur compound from garlic, on selective accumulation of the ion in the plasma membrane fraction via allicin-mediated phospholipid peroxidation. Planta Med. 2006;72:875–80.CrossRefPubMedGoogle Scholar
  35. 35.
    Ledezmal E, Maniscalchi MT, Espinoza DL. Synergy between ajoene and ketoconazole in isolates of Microsporum canis. Rev Iberoam Micol. 2008;25:157–62.CrossRefGoogle Scholar
  36. 36.
    Pfaller MA, Boyken LB, Hollis RJ, Kroeger J, Messer SA, Tendolkar S, et al. Validation of 24-hour fluconazole MIC readings versus the CLSI 48-hour broth microdilution reference method: results from a global Candida antifungal surveillance programme. J Clin Microbiol. 2008;46(11):3585–90.CrossRefPubMedGoogle Scholar
  37. 37.
    Lozano-Chiu M, Arikan S, Paetznick VL, Anaissie EJ, Rex JH. Optimizing voriconazole susceptibility testing of Candida: effects of incubation time, endpoint rule, species of Candida, and level of fluconazole susceptibility. J Clin Microbiol. 1999;37:2755–9.PubMedGoogle Scholar
  38. 38.
    Revankar SG, Kirkpatrick WR, McAtee RK, Fothergill AW, Redding SW, Rinaldi MG, et al. Interpretation of trailing endpoints in antifungal susceptibility testing by the National Committee for Clinical Laboratory Standards method. J Clin Microbiol. 1998;36:153–6.PubMedGoogle Scholar
  39. 39.
    Barry AL, Pfaller MA, Rennie RP, Fuchs PC, Brown SD. Precision and accuracy of fluconazole susceptibility testing by broth microdilution, E-test, and disk diffusion methods. Antimicrob Agents Chemother. 2002;46(6):1781–4.CrossRefPubMedGoogle Scholar
  40. 40.
    Freeman F, Kodera Y. Garlic chemistry: stability of S-(2-propenyl)-2-propene-1- sulfinothionate (allicin) in blood, solvents, and simulated physiological fluids. J Agric Food Chem. 1995;43:2332–8.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Alireza Khodavandi
    • 1
  • Fahimeh Alizadeh
    • 2
  • Farzad Aala
    • 3
  • Zamberi Sekawi
    • 4
  • Pei Pei Chong
    • 1
    • 5
  1. 1.Department of Biomedical Sciences, Faculty of Medicine and Health SciencesUniversiti Putra MalaysiaSerdangMalaysia
  2. 2.Institute of Tropical AgricultureUniversiti Putra MalaysiaSerdangMalaysia
  3. 3.Department of Biology, Faculty of ScienceUniversiti Putra MalaysiaSerdangMalaysia
  4. 4.Department of Medical Microbiology and Parasitology, Faculty of Medicine and Health SciencesUniversiti Putra MalaysiaSerdangMalaysia
  5. 5.Institute of BioscienceUniversiti Putra MalaysiaSerdangMalaysia

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