{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,21]],"date-time":"2026-09-21T20:20:26Z","timestamp":1790022026730,"version":"4.0.1"},"reference-count":39,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2025,9,3]],"date-time":"2025-09-03T00:00:00Z","timestamp":1756857600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004281","name":"National Science Center (NCN)","doi-asserted-by":"publisher","award":["2021\/43\/B\/NZ1\/00523"],"award-info":[{"award-number":["2021\/43\/B\/NZ1\/00523"]}],"id":[{"id":"10.13039\/501100004281","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Eugenol is a compound with promising antimicrobial properties. The rising phenomenon of multidrug resistance of Candida albicans is driving researchers to search for new, alternative therapeutics that would synergize with conventional antifungal drugs. The aim of the present study is to investigate how eugenol and eugenol-based extracts impair C. albicans growth by generation of reactive oxygen species (ROS) and plasma membrane (PM) disruption. The methods that we applied involve structural analysis of eugenol extracts by HPLC, ATR-FTIR, and polyphenol detection. Additionally, determination of ROS level in C. albicans was performed using microscopic and flow cytometry studies and analysis of PM integrity (PI-staining, observation of PM transporter\u2014Cdr1p\u2014localization) and fluidity (fluorometric study). The results indicate that eugenol impacts fungal growth, and this corresponds with increased ROS levels and diminished PM fluidity in the C. albicans WT strain. C. albicans strains deprived of ergosterol (erg11\u0394\/\u0394) exhibited lowered ROS level and no change in PM fluidity in response to the tested eugenol extracts, but they affected its growth and caused PM permeabilization and Cdr1p delocalization. These conclusions indicate that mode of action of eugenol can be related to disruption of PM structure by both ergosterol-dependent and -independent mechanisms. Ergosterol can play a crucial role in maintaining the PM integrity during treatment with lower concentrations of eugenol.<\/jats:p>","DOI":"10.3390\/ijms26178571","type":"journal-article","created":{"date-parts":[[2025,9,4]],"date-time":"2025-09-04T09:18:57Z","timestamp":1756977537000},"page":"8571","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Multifunctional Activity of Syzygium aromaticum Extracts Against Candida albicans: Free Radicals, Membrane Permeabilization and Cdr1p Localization"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9629-9451","authenticated-orcid":false,"given":"Daria","family":"Derkacz","sequence":"first","affiliation":[{"name":"Department of Biotransformation, Faculty of Biotechnology, University of Wroclaw, F. Joliot-Curie 14A, 50-383 Wroclaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-6002-0601","authenticated-orcid":false,"given":"Liliana","family":"Cebula","sequence":"additional","affiliation":[{"name":"Department of Biotransformation, Faculty of Biotechnology, University of Wroclaw, F. Joliot-Curie 14A, 50-383 Wroclaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1925-193X","authenticated-orcid":false,"given":"Anna","family":"Krasowska","sequence":"additional","affiliation":[{"name":"Department of Biotransformation, Faculty of Biotechnology, University of Wroclaw, F. Joliot-Curie 14A, 50-383 Wroclaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Bezerra, D.P., Milit\u00e3o, G.C.G., de Morais, M.C., and de Sousa, D.P. (2017). The Dual Antioxidant\/Prooxidant Effect of Eugenol and Its Action in Cancer Development and Treatment. Nutrients, 9.","DOI":"10.3390\/nu9121367"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"2497354","DOI":"10.1155\/2021\/2497354","article-title":"Pharmacological Properties and Health Benefits of Eugenol: A Comprehensive Review","volume":"2021","author":"Nisar","year":"2021","journal-title":"Oxidative Med. Cell. Longev."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Ulanowska, M., and Olas, B. (2021). Biological Properties and Prospects for the Application of Eugenol\u2014A Review. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22073671"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"975","DOI":"10.1089\/jmf.2010.0197","article-title":"Antioxidant activity of eugenol: A structure-activity relationship study","volume":"14","year":"2011","journal-title":"J. Med. Food"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"bcr2013009464","DOI":"10.1136\/bcr-2013-009464","article-title":"An unexpected positive hypersensitive reaction to eugenol","volume":"2013","author":"Tammannavar","year":"2013","journal-title":"BMJ Case Rep."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Didehdar, M., Chegini, Z., and Shariati, A. (2022). Eugenol: A novel therapeutic agent for the inhibition of Candida species infection. Front. Pharmacol., 13.","DOI":"10.3389\/fphar.2022.872127"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1124","DOI":"10.4315\/0362-028X-73.6.1124","article-title":"Antifungal Activity of Eugenol against Penicillium, Aspergillus, and Fusarium Species","volume":"73","author":"Campaniello","year":"2010","journal-title":"J. Food Prot."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"100406","DOI":"10.1016\/j.hermed.2020.100406","article-title":"Mechanisms of bactericidal action of Eugenol against Escherichia coli","volume":"26","author":"Jeyakumar","year":"2020","journal-title":"J. Herb. Med."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2821","DOI":"10.1111\/jam.15171","article-title":"Antibiofilm and anti-quorum sensing activities of eugenol and linalool from Ocimum tenuiflorum against Pseudomonas aeruginosa biofilm","volume":"131","author":"Lahiri","year":"2021","journal-title":"J. Appl. Microbiol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1178","DOI":"10.1099\/jmm.0.020693-0","article-title":"In vitro synergy of eugenol and methyleugenol with fluconazole against clinical Candida isolates","volume":"59","author":"Ahmad","year":"2010","journal-title":"J. Med. Microbiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"105456","DOI":"10.1016\/j.micpath.2022.105456","article-title":"Antibacterial activity of eugenol on the IS-58 strain of Staphylococcus aureus resistant to tetracycline and toxicity in Drosophila melanogaster","volume":"164","author":"Bezerra","year":"2022","journal-title":"Microb. Pathog."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"115436","DOI":"10.1016\/j.ejmech.2023.115436","article-title":"New miconazole-based azoles derived from eugenol show activity against Candida spp. and Cryptococcus gattii by inhibiting the fungal ergosterol biosynthesis","volume":"256","author":"Reis","year":"2023","journal-title":"Eur. J. Med. Chem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/j.micpath.2009.10.001","article-title":"Evolution of ergosterol biosynthesis inhibitors as fungicidal against Candida","volume":"48","author":"Ahmad","year":"2010","journal-title":"Microb. Pathog."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1186\/s12906-020-2818-8","article-title":"In vitro anticandidal potency of Syzygium aromaticum (clove) extracts against vaginal candidiasis","volume":"20","author":"Yassin","year":"2020","journal-title":"Complement. Med. Ther."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Ahmed, I.A.M., Babiker, E.E., Al-Juhaimi, F.Y., and Bekhit, A.E.A. (2022). Clove Polyphenolic Compounds Improve the Microbiological Status, Lipid Stability, and Sensory Attributes of Beef Burgers during Cold Storage. Antioxidants, 11.","DOI":"10.3390\/antiox11071354"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"201","DOI":"10.4274\/tjps.62207","article-title":"Pharmacological and Toxicological Properties of Eugenol","volume":"14","year":"2017","journal-title":"Turk. J. Pharm. Sci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"767","DOI":"10.1080\/10610278.2019.1702663","article-title":"Mechanism for efficient separation of eugenol and eugenol acetate with \u03b2-cyclodextrin as a selective solvent","volume":"31","author":"Yang","year":"2019","journal-title":"Supramol. Chem."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Batiha, G.E., Alkazmi, L.M., Wasef, L.G., Beshbishy, A.M., Nadwa, E.H., and Rashwan, E.K. (2020). Syzygium aromaticum L. (Myrtaceae): Traditional Uses, Bioactive Chemical Constituents, Pharmacological and Toxicological Activities. Biomolecules, 10.","DOI":"10.3390\/biom10020202"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Shahina, Z., Ndlovu, E., Persaud, O., Sultana, T., and Dahms, T.E.S. (2022). Candida albicans Reactive Oxygen Species (ROS)-Dependent Lethality and ROS-Independent Hyphal and Biofilm Inhibition by Eugenol and Citral. Microbiol. Spectr., 10.","DOI":"10.1128\/spectrum.03183-22"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Biernasiuk, A., Baj, T., and Malm, A. (2022). Clove Essential Oil and Its Main Constituent, Eugenol, as Potential Natural Antifungals against Candida spp. Alone or in Combination with Other Antimycotics Due to Synergistic Interactions. Molecules, 28.","DOI":"10.3390\/molecules28010215"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"5248","DOI":"10.1111\/1462-2920.16157","article-title":"Transcriptional regulation of ergosterol biosynthesis genes in response toiron deficiency","volume":"24","author":"Alepuz","year":"2022","journal-title":"Environ. Microbiol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"579","DOI":"10.3390\/jof9050579","article-title":"Essential Role of CgErg6p in Maintaining Oxidative Stress Tolerance and Iron Homeostasis in Candida glabrata","volume":"9","author":"Elias","year":"2023","journal-title":"J. Fungi"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1454","DOI":"10.1099\/jmm.0.010538-0","article-title":"Antifungal activity of the clove essential oil from Syzygium aromaticum on Candida, Aspergillus and dermatophyte species","volume":"58","author":"Pinto","year":"2009","journal-title":"J. Med. Microbiol."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Kowalewska, A., and Majewska-Smolarek, K. (2023). Eugenol-Based Polymeric Materials-Antibacterial Activity and Applications. Antibiotics, 12.","DOI":"10.3390\/antibiotics12111570"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"378","DOI":"10.3390\/microorganisms7100378","article-title":"A Crucial Role for Ergosterol in Plasma Membrane Composition, Localisation, and Activity of Cdr1p and H+-ATPase in Candida albicans","volume":"7","author":"Suchodolski","year":"2019","journal-title":"Microorganism"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"505204","DOI":"10.1155\/2014\/505204","article-title":"Effect of Eugenol on Cell Surface Hydrophobicity, Adhesion, and Biofilm of Candida tropicalis and Candida dubliniensis Isolated from Oral Cavity of HIV-Infected Patients","volume":"2014","author":"Bartelli","year":"2014","journal-title":"Evid.-Based Complement. Altern. Med."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Ahmad, A., Wani, M.Y., Khan, A., Manzoor, N., and Molepo, J. (2015). Synergistic Interactions of Eugenol-tosylate and Its Congeners with Fluconazole against Candida albicans. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0145053"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Prajapati, J., Goswami, D., Dabhi, M., Acharya, D., and Rawal, R.M. (2022). Potential dual inhibition of SE and CYP51 by eugenol conferring inhibition of Candida albicans: Computationally curated study with experimental validation. Comput. Biol. Med., 151.","DOI":"10.1016\/j.compbiomed.2022.106237"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"113303","DOI":"10.1016\/j.lwt.2022.113303","article-title":"Antifungal activity of thymol against the main fungi causing pomegranate fruit rot by suppressing the activity of cell wall degrading enzymes","volume":"161","author":"Ranjbar","year":"2022","journal-title":"LWT"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"6953","DOI":"10.3390\/molecules17066953","article-title":"Eugenol\u2014From the remote Maluku Islands to the international market place: A review of a remarkable and versatile molecule","volume":"17","author":"Kamatou","year":"2012","journal-title":"Molecules"},{"key":"ref_31","first-page":"96","article-title":"HPLC\u2013UV method development and quantification of eugenol from methanolic extracts of some spices","volume":"3","author":"Inam","year":"2014","journal-title":"Int. J. Chem. Phys. Sci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"717","DOI":"10.1093\/genetics\/134.3.717","article-title":"Isogenic strain construction and gene mapping in Candida albicans","volume":"134","author":"Fonzi","year":"1993","journal-title":"Genetics"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Szczepaniak, J., \u0141ukaszewicz, M., and Krasowska, A. (2015). Estimation of Candida albicans ABC Transporter Behavior in Real-Time via Fluorescence. Front. Microbiol., 6.","DOI":"10.3389\/fmicb.2015.01382"},{"key":"ref_34","unstructured":"NCCLS (2004). Method for Antifungal Disk Diffusion Susceptibility Testing of Yeasts; Approved Guideline. NCCLS Document M44-A, NCCLS."},{"key":"ref_35","unstructured":"(2008). Reference Method for Broth Dilution Antifungal Susceptibility Testing of Yeast (Standard No. CLSI M27-A3 28)."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Urbanek, A.K., Muraszko, J., Derkacz, D., \u0141ukaszewicz, M., Bernat, P., and Krasowska, A. (2022). The Role of Ergosterol and Sphingolipids in the Localization and Activity of Candida albicans\u2019 Multidrug Transporter Cdr1p and Plasma Membrane ATPase Pma1p. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23179975"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Derkacz, D., Bernat, P., and Krasowska, A. (2022). K143R Amino Acid Substitution in 14-\u03b1-Demethylase (Erg11p) Changes Plasma Membrane and Cell Wall Structure of Candida albicans. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23031631"},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Derkacz, D., Grzybowska, M., Cebula, L., and Krasowska, A. (2023). Surfactin and Capric Acid Affect the Posaconazole Susceptibility of Candida albicans Strains with Altered Sterols and Sphingolipids Biosynthesis. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms242417499"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Ishmayana, S., Kennedy, U.J., and Learmonth, R.P. (2017). Further investigation of relationships between membrane fluidity and ethanol tolerance in Saccharomyces cerevisiae. World J. Microbiol. Biotechnol., 33.","DOI":"10.1007\/s11274-017-2380-9"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/17\/8571\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:38:53Z","timestamp":1760035133000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/17\/8571"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,9,3]]},"references-count":39,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2025,9]]}},"alternative-id":["ijms26178571"],"URL":"https:\/\/doi.org\/10.3390\/ijms26178571","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,9,3]]}}}