Chemical Characterization of Lavandula dentata Essential Oil Cultivated in Chile and Its Antibiofilm Effect against Candida albicans

Abstract

Candida albicans is the most common human fungal pathogen, and with the increase in resistance rates worldwide, it is necessary to search for new pharmacological alternatives. Lavandula dentata L. essential oil is recognized as having antimicrobial properties. However, its effect against fungal biofilms has been poorly described. C. albicans-related infections involve the development of biofilms, which are highly resistant to conventional antifungals. In this work, we evaluated the antibiofilm effect of L. dentata L. essential oil against C. albicans. First, we characterized the essential oil by gas chromatography-mass spectrometry. The antifungal effect on C. albicans reference strains was evaluated by a disk diffusion assay and the minimal inhibitory concentration was obtained through a microdilution assay. The effect of the essential oil on the adhesion ability of C. albicans was determined through a crystal violet assay, and morphogenesis inhibition was assessed by light microscopy. The effect of the essential oil on the microarchitecture of biofilms was evaluated through scanning electron microscopy. Finally, the antibiofilm effect was evaluated through an adapted biofilm scratch assay and XTT viability assay. The main constituent of the essential oil was the monoterpenoid eucalyptol (60%). The essential oil presented minimal inhibitory concentrations of 156 and 130 µg/mL against two strains assayed. This minimal inhibitory concentration inhibited adhesion, morphogenesis, biofilm formation, altered microarchitecture, and decreased the viability of established biofilms formed on abiotic surfaces for both strains assayed. This study demonstrates that the essential oil from L. dentata could be a promising treatment against C. albicans biofilms.

Key words

Lavandula dentate - Lamiaceae - essential oil - antifungal - antibiofilm - Candida albicans

• References

• 1 Fisher MC, Hawkins NJ, Sanglard D, Gurr SJ. Worldwide emergence of resistance to antifungal drugs challenges human health and food security. Science 2018; 360: 739-742
• 2 Perfect JR. The antifungal pipeline: a reality check. Nat Rev Drug Discov 2017; 16: 603-616
• 3 Dadar M, Tiwari R, Karthik K, Chakraborty S, Shahali Y, Dhama K. Candida albicans – Biology, molecular characterization, pathogenicity, and advances in diagnosis and control – An update. Microb Pathog 2018; 117: 128-138
• 4 Hidron AI, Edwards JR, Patel J, Horan TC, Sievert DM, Pollock DA, Fridkin SK. NHSN: annual report: antimicrobial-resistant pathogens associated with healthcare-associated infections: annual summary of data reported to the National Healthcare Safety Network at the Centers for Disease Control and Prevention, 2006–2007. Infect Control Hosp Epidemiol 2008; 29: 996-1011
• 5 Pfaller M, Neofytos D, Diekema D, Azie N, Meier-Kriesche HU, Quan SP, Horn D. Epidemiology and outcomes of candidemia in 3648 patients: data from the Prospective Antifungal Therapy (PATH Alliance) registry, 2004–2008. Diagn Microbiol Infect Dis 2012; 74: 323-331
• 6 Kim J, Sudbery P. Candida albicans, a major human fungal pathogen. J Microbiol 2011; 49: 171-177
• 7 Cuéllar-Cruz M, López-Romero E, Villagómez-Castro J, Ruiz-Baca E. Candida species: new insights into biofilm formation. Future Microbiol 2012; 7: 755-771
• 8 Lohse MB, Gulati M, Johnson AD, Nobile CJ. Development and regulation of single-and multi-species Candida albicans biofilms. Nat Rev Microbiol 2018; 16: 19-31
• 9 Silva S, Rodrigues C, Araújo D, Rodrigues M, Henriques M. Candida species biofilmsʼ antifungal resistance. J Fungi 2017; 3: e8
• 10 Cannon RD, Holmes AR. Learning the ABC of oral fungal drug resistance. Mol Oral Microbiol 2015; 30: 425-437
• 11 Ramage G, Bachmann S, Patterson TF, Wickes BL, López-Ribot JL. Investigation of multidrug efflux pumps in relation to fluconazole resistance in Candida albicans biofilms. J Antimicrob Chemother 2002; 49: 973-980
• 12 Tobudic S, Kratzer C, Lassnigg A, Presterl E. Antifungal susceptibility of Candida albicans in biofilms. Mycoses 2012; 55: 199-204
• 13 Kong E, Tsui C, Kucharikova S, Andes D, Van Dijck P, Jabra-Rizk M. Commensal protection of Staphylococcus aureus against antimicrobials by Candida albicans biofilm matrix. MBio 2016; 7: 1-12
• 14 Mittal RP, Rana A, Jaitak V. Essential oils: an impending substitute of synthetic antimicrobial agents to overcome antimicrobial resistance. Curr Drug Targets 2019; 20: 605-624
• 15 Gao J, Wang H, Li Z, Wong AHH, Wang YZ, Guo Y, Lin X, Zeng G, Wang Y, Wang J. Candida albicans gains azole resistance by altering sphingolipid composition. Nat Commun 2018; 9: 1-14
• 16 Justus B, de Almeida VP, Gonçalves MM, de Assunção DPSF, Borsato DM, Arana AFM, Maia BHLNS, de Paula JFP, Budel JM, Farago PV. Chemical composition and biological activities of the essential oil and anatomical markers of Lavandula Dentata L. Cultivated In Brazil. Brazilian Arch Biol Technol 2018; 61: e18180111
• 17 Zuzarte M, Gonçalves MJ, Cavaleiro C, Cruz MT, Benzarti A, Marongiu B, Maxia A, Piras A, Salgueiro L. Antifungal and anti-inflammatory potential of Lavandula stoechas and Thymus herba-barona essential oils. Ind Crops Prod 2013; 44: 97-103
• 18 Budzyñska A, Wiêckowska-Szakiel M, Sadowska B, Kalemba D, Rozalka B. Antibiofilm activity of selected plant essential oils and their major components. Polish J Microbiol 2011; 60: 35-41
• 19 Bouazama S, Harhar H, Costa J, Desjobert JM, Talbaoui A, Tabyaoui M. Chemical composition and antibacterial activity of the essential oils of Lavandula pedunculata and Lavandula dentata . J Mater Environ Sci 2017; 8: 2154-2160
• 20 Dudai N. Factors affecting content and composition of essential oils in aromatic plants. Helsinki: WLF Publisher; 2005
• 21 Morcia C, Malnati M, Terzi V. In vitro antifungal activity of terpinen-4-ol, eugenol, carvone, 1, 8-cineole (eucalyptol) and thymol against mycotoxigenic plant pathogens. Food Addit Contam 2012; 29: 415-422
• 22 Rivas da Silva AC, Monteiro Lopes P, Barros de Azevedo MM, Machado Costa DC, Sales Alviano C, Sales Alviano D. Biological activities of α-pinene and β-pinene enantiomers. Molecules 2012; 17: 6305-6316
• 23 Kifer D, Mužinić V, Klaric MŠ. Antimicrobial potency of single and combined mupirocin and monoterpenes, thymol, menthol and 1,8-cineole against Staphylococcus aureus planktonic and biofilm growth. J Antibiot (Tokyo) 2016; 69: 689-696
• 24 Cavalheiro M, Teixeira MC. Candida Biofilms: Threats, Challenges, and Promising Strategies. Front Med 2018; 5: 1-15
• 25 Bassyouni RH, Wali IE, Kamel Z, Kassim MF. Fennel oil: A promising antifungal agent against biofilm forming fluconazole resistant Candida albicans causing vulvovaginal candidiasis. J Herb Med 2018; 5: 1-5
• 26 Raut JS, Shinde RB, Chauhan NM, Mohan Karuppayil S. Terpenoids of plant origin inhibit morphogenesis, adhesion, and biofilm formation by Candida albicans . Biofouling 2013; 29: 87-96
• 27 Van Acker H, Van Dijck P, Coenye T. Molecular mechanisms of antimicrobial tolerance and resistance in bacterial and fungal biofilms. Trends Microbiol 2014; 22: 326-333
• 28 Zore GB, Thakre AD, Jadhav S, Karuppayil SM. Terpenoids inhibit Candida albicans growth by affecting membrane integrity and arrest of cell cycle. Phytomedicine 2011; 18: 1181-1190
• 29 Haegler P, Joerin L, Krähenbühl S, Bouitbir J. Hepatocellular toxicity of imidazole and triazole antimycotic agents. Toxicol Sci 2017; 157: 183-195
• 30 Pierce CG, Uppuluri P, Tristan AR, Wormley FL, Mowat E, Ramage G, Lopez-Ribot JL. A simple and reproducible 96-well plate-based method for the formation of fungal biofilms and its application to antifungal susceptibility testing. Nat Protoc 2008; 3: 1494-1500
• 31 Madariaga-Venegas F, Ferna R, Duarte LF, Suarez N, Delgadillo D, Jara JA, Fernández-Ramires R, Urzúa B, Molina-Berríos A. Characterization of a novel antibiofilm effect of nitric oxide-releasing aspirin (NCX-4040) on Candida albicans isolates from denture stomatitis patients. PLoS One 2017; 12: e0176755