Antifungal Activity of the Essential Oil of Thymus x viciosoi against Candida, Cryptococcus, Aspergillus and Dermatophyte Species

Abstract

The essential oil (EO) of Thymus x viciosoi (Pau) R. Morales was isolated and analysed by GC and GC‐MS. The antifungal activity of the EO and its major components against clinically relevant yeasts and molds was then measured. Their influence on the germ tube formation in Candida albicans and the influence of the EO on the metabolic function and cytoplasmic membrane integrity in the same yeast, analyzed by flow cytometry, were also studied. The EO showed high contents of carvacrol, thymol, and p-cymene. The total EO, as well as its components carvacrol and thymol, displayed very low minimum inhibitory concentrations and minimum fungicidal concentrations against all tested organisms (0.04 to 0.64 µL · mL⁻¹), while p-cymene showed weaker activity (2.5 to > 20.0 µL · mL⁻¹). They also inhibited filamentation at sub-inhibitory concentrations in C. albicans, particularly p-cymene, and the EO led to rapid metabolic arrest, disruption of the plasma membrane and consequently cell death. The EO and its main components were found to display a broad fungicidal activity through the disruption of cytoplasmic membrane integrity leading to leakage of vital intracellular compounds. In conclusion, the phenolic oil of T. x viciosoi may have potential for use in the development of clinically useful antifungal preparations.

Thymus x viciosoi (Pau) R. Morales - Lamiaceae - phenolic oil - antifungal activity - Candida albicans - flow cytometry - cytoplasmic membrane disruption

References

• 1 Richardson M, Lass-Florl C. Changing epidemiology of systemic fungal infections.  Clin Microbiol Infect. 2008;  14 (Suppl. 4) 5-24
• 2 Edmond M B, Wallace S E, McClish D K, Pfaller M A, Jones R N, Wenzel R P. Nosocomial bloodstream infections in United States hospitals: a three-year analysis.  Clin Infect Dis. 1999;  29 239-244
• 3 Eggimann P, Garbino J, Pittet D. Epidemiology of Candida species infections in critically ill non-immunosuppressed patients.  Lancet Infect Dis. 2003;  3 685-702
• 4 Gupta A K, Cooper E A. Update in antifungal therapy of dermatophytosis.  Mycopathologia. 2008;  166 353-367
• 5 de Pauw B E, Picazo J J. Present situation in the treatment of invasive fungal infection.  Int J Antimicrob Agents. 2008;  32 (Suppl. 2) S167-S171
• 6 Pauli A. Anticandidal low molecular compounds from higher plants with special reference to compounds from essential oils.  Med Res Rev. 2006;  26 223-268
• 7 Edris A E. Pharmaceutical and therapeutic potentials of essential oils and their individual volatile constituents: a review.  Phytother Res. 2007;  21 308-323
• 8 Kalemba D, Kunicka A. Antibacterial and antifungal properties of essential oils.  Curr Med Chem. 2003;  10 813-829
• 9 Bakkali F, Averbeck S, Averbeck D, Idaomar M. Biological effects of essential oils – a review.  Food Chem Toxicol. 2008;  46 446-475
• 10 Figueiredo A C, Barroso J G, Pedro L G, Salgueiro L, Miguel M G, Faleiro M L. Portuguese Thymbra and Thymus species volatiles: chemical composition and biological activities.  Curr Pharm Des. 2008;  14 3120-3140
• 11 Pina-Vaz C, Rodrigues A G, Pinto E, Costa-de-Oliveira S, Tavares C, Salgueiro L, Cavaleiro C, Gonçalves M J, Martinez-de-Oliveira J. Antifungal activity of Thymus oils and their major compounds.  J Eur Acad Dermatol Venereol. 2004;  18 73-78
• 12 Pinto E, Pina-Vaz C, Salgueiro L, Gonçalves M J, Costa-de-Oliveira S, Cavaleiro C, Palmeira A, Rodrigues A, Martinez-de-Oliveira J. Antifungal activity of the essential oil of Thymus pulegioides on Candida, Aspergillus and dermatophyte species.  J Med Microbiol. 2006;  55 1367-1373
• 13 Salgueiro L R, Pinto E, Goncalves M J, Costa I, Palmeira A, Cavaleiro C, Pina-Vaz C, Rodrigues A G, Martinez-de-Oliveira J. Antifungal activity of the essential oil of Thymus capitellatus against Candida, Aspergillus and dermatophyte strains.  Flavour Fragrance J. 2006;  21 749-753
• 14 Sokovic M, Glamoclija J, Ciric A, Kataranovski D, Marin P D, Vukojevic J, Brkic D. Antifungal activity of the essential oil of Thymus vulgaris L. and thymol on experimentally induced dermatomycoses.  Drug Dev Ind Pharm. 2008;  34 1388-1393
• 15 das Neves J, Pinto E, Teixeira B, Dias G, Rocha P, Cunha T, Santos B, Amaral M H, Bahia M F. Local treatment of vulvovaginal candidosis – general and practical considerations.  Drugs. 2008;  68 1787-1802
• 16 Saville S P, Lazzell A L, Bryant A P, Fretzen A, Monreal A, Solberg E O, Monteagudo C, Lopez-Ribot J L, Milne G T. Inhibition of filamentation can be used to treat disseminated candidiasis.  Antimicrob Agents Chemother. 2006;  50 3312-3316
• 17 Adams R P. Identification of essential oil components by gas chromatography/mass spectroscopy. Carol Stream; Allured Publishing Corporation 2004
• 18 Joulain D, Konig W A. The atlas of spectral data of sesquiterpene hydrocarbons. Hamburg; E. B. – Verlag Hamburg 1998
• 19 Clinical and Laboratory Standards Institute (CLSI) .Reference method for broth dilution antifungal susceptibility testing of yeasts; Approved standard, third edition M27-A3. Wayne; Clinical and Laboratory Standards Institute 2008
• 20 Clinical and Laboratory Standards Institute (CLSI) .Reference method for broth dilution antifungal susceptibility testing of yeasts, third informational supplement M27-S3. Wayne; Clinical and Laboratory Standards Institute 2008
• 21 Clinical and Laboratory Standards Institute (CLSI) .Reference method for broth dilution antifungal susceptibility testing of filamentous fungi; approved standard, second edition M38-A2. Wayne; Clinical and Laboratory Standards Institute 2008
• 22 Marichal P, Gorrens J, Vancutsem J, Vandenbossche H. Culture media for the study of the effects of azole derivatives on germ tube formation and hyphal growth of Candida albicans.  Mykosen. 1986;  29 76-81
• 23 Mitchell A P. Dimorphism and virulence in Candida albicans.  Curr Opin Microbiol. 1998;  1 687-692
• 24 Millard P J, Roth B L, Thi H P, Yue S T, Haugland R P. Development of the FUN-1 family of fluorescent probes for vacuole labeling and viability testing of yeasts.  Appl Environ Microbiol. 1997;  63 2897-2905
• 25 Pinto E, Queiroz M J, Vale-Silva L A, Oliveira J F, Begouin A, Begouin J M, Kirsch G. Antifungal activity of synthetic di(hetero)arylamines based on the benzo[b]thiophene moiety.  Bioorg Med Chem. 2008;  16 8172-8177