Uniwersytet Medyczny w Łodzi Medical University of Lodz https://publicum.umed.lodz.pl Inhibitory Effect of Eugenol and Trans-Anethole Alone and in Combination with Antifungal Medicines on Candida albicans Clinical Isolates, Publikacja / Publication Dąbrowska Marta, Zielińska-Bliźniewska Hanna, Kwiatkowski Paweł, Łopusiewicz Łukasz, Prus Agata, Kostek Mateusz, Kochan Ewa, Sienkiewicz Monika DOI wersji wydawcy / Published http://dx.doi.org/10.1002/cbdv.202000843 version DOI Adres publikacji w Repozytorium URL / Publication address in https://publicum.umed.lodz.pl/info/article/AML3aacc66138914656916b7a87814195e4/ Repository Data opublikowania w Repozytorium 2021-03-27 / Deposited in Repository on Rodzaj licencji / Type of licence Other open licence Dąbrowska Marta, Zielińska-Bliźniewska Hanna, Kwiatkowski Paweł, Łopusiewicz Łukasz, Prus Agata, Kostek Mateusz, Kochan Ewa, Sienkiewicz Monika: Inhibitory Cytuj tę wersję / Cite this version Effect of Eugenol and Trans-Anethole Alone and in Combination with Antifungal Medicines on Candida albicans Clinical Isolates, Chemistry & Biodiversity, vol. 18, no. 5, 2021, pp. 1-11, DOI:10.1002/cbdv.202000843 Title: Inhibitory Effect of Eugenol and Trans-Anethole Alone and in Combination with Antifungal Medicines on Candida albicans Clinical Isolates Authors: Marta Dąbrowska, Hanna Zielińska-Bliźniewska, Paweł Kwiatkowski, Łukasz Łopusiewicz, Agata Pruss, Mateusz Kostek, Ewa Kochan, and Monika Sienkiewicz This manuscript has been accepted after peer review and appears as an Accepted Article online prior to editing, proofing, and formal publication of the final Version of Record (VoR). This work is currently citable by using the Digital Object Identifier (DOI) given below. The VoR will be published online in Early View as soon as possible and may be different to this Accepted Article as a result of editing. Readers should obtain the VoR from the journal website shown below when it is published to ensure accuracy of information. The authors are responsible for the content of this Accepted Article. To be cited as: Chem. Biodiversity 10.1002/cbdv.202000843 Link to VoR: https://doi.org/10.1002/cbdv.202000843 Pobrano z https://publicum.umed.lodz.pl / Downloaded from Repository of Medical University of Lodz 2021-10-07 Chemistry & Biodiversity 10.1002/cbdv.202000843 Chem. Biodiversity Inhibitory Effect of Eugenol and Trans-Anethole Alone and in Combination with Antifungal Medicines on Candida albicans Clinical Isolates Marta Dąbrowskaa, Hanna Zielińska-Bliźniewskaa, Paweł Kwiatkowskib, Łukasz Łopusiewiczc, Agata Prussd, Mateusz Kostekc, Ewa Kochane, Monika Sienkiewicza* a Department of Allergology and Respiratory Rehabilitation, Medical University of Lodz, Żeligowskiego 7/9 Str., 90-752 Lodz, Poland; [email protected] b Department of Diagnostic Immunology; Chair of Microbiology, Immunology and Laboratory Medicine; Pomeranian Medical University in Szczecin; 72 Powstańców Wielkopolskich Avenue; 70-111 Szczecin, Poland c Center of Bioimmobilisation and Innovative Packaging Materials; Faculty of Food Sciences and Fisheries; West Pomeranian University of Technology in Szczecin; Janickiego 35; 71-270 Szczecin, Poland d Department of Laboratory Medicine; Chair of Microbiology, Immunology and Laboratory Medicine; Pomeranian Medical University in Szczecin; 72 Powstańców Wielkopolskich Avenue; 70-111 Szczecin, Poland e Pharmaceutical Biotechnology Department, Medical University of Lodz, Muszyńskiego 1, 90-151 Łódź, Poland Abstract: One of the most common pathogens among yeasts is Candida albicans, which presents a serious health threat. The study aimed to check the antifungal properties of trans-anethole and eugenol with selected antifungal medicines (AMs) against C. albicans clinical isolates. The checkerboard method was used to tests of interactions between these compounds. Achieved results indicated that eugenol showed synergistic and additive activities with miconazole and econazole against investigated clinical isolates, respectively. Moreover, the combination — trans-anethole — miconazole also showed an additive effect against two clinical isolate. We tried to relate the results to changes in C. albicans cell sheaths under the influence of essential oils compounds (EOCs) performing the Fourier transform infrared spectra analysis to confirm the presence of particular chemical moieties in C. albicans cells. Nevertheless, no strong relationships was observed between synergistic and additive actions of used EOC-AMs combinations and chemical moieties in C. albicans cells. Keywords: Candida spp., trans-anethole, eugenol, synergistic activity, miconazole Introduction Vulvovaginal candidiasis (VVC) is the second most common abnormality associated with vaginal biocenosis.[1] In most cases, there are endogenous infections, and the reason for the disease process is the disturbance of the balance between the yeast and the host.[2] It is estimated that about 75% of women will experience at least one episode of VVC over a lifetime, and approximately 40-45% will have two or more episodes. In 5-10% of patients, the recurrent form of mycosis will develop. Asymptomatic colonization affects 10-15% of women. [3] According to available data, the most common pathogen responsible for VVC is Candida albicans.[4] Nevertheless, complicated VVC is defined as a severe or recurrent disease (more than four episodes of symptomatic VVC within one year), infection due to Candida species other than [5] C. albicans, and/or VVC in an abnormal host. Several topical antifungal agents are effective therapy for VVC, and no agent is clearly Manuscript superior.[6,7] Oral and topical antimycotics achieve entirely equivalent results.[8] For recurring VVC, therapy with a topical or oral azole, followed by fluconazole is recommended.[9] The literature showed that several mechanisms were found to fluconazole resistance among Candida's genus, and an increase in azole resistance may have a clinical impact.[10] The evolution and increasing factor of antimicrobial resistance in pathogens have prompted extensive research to find alternative therapeutics; for example, some essential oils (EOs) and their main compounds.[11] EOs obtained from lemongrass (Cymbopogon citratus), geranium (Pelargonium asperum),[12] tea tree (Melaleuca alternifolia),[13,14] lavender (Lavandula angustifolia),[15] and thyme (Thymus vulgaris)[16] are reported to inhibit Candida mycelial growth in vitro. Pietrella et al.[17], show that the EO of a Moroccan plant Mentha suaveolens is candidastatic and candidacidal and has a degree of anticandidal activity in a model of vaginal infection. In vivo studies are very limited; Maruyama et al.[18], examine the efficacy of vaginal application of geranium EO, or its main compound, geraniol, against murine experimental candidiasis was investigated. In the study, there was shown that the application of geranium oil or its component, geraniol, suppressed Candida cell growth in the vaginal cavity only when it was combined with vaginal washing. Natural bioactive substances can be used in combination therapy. Mertas et al.[19], showed that combining natural substances such as tea tree EO and conventional drugs such as fluconazole may help treat difficult yeast infections. In our research, we focused on examining the antifungal properties of trans-anethole and eugenol against C. albicans clinical isolates come from vaginal secretions. Both oil components show biological activity and seem to be promising ingredients in the fight against pathogenic microorganisms, especially those showing resistance to standard medications in medical practice.[20,21] Therefore, the study aimed to assess the antifungal activity of trans-anethole and eugenol alone and in combination with selected antifungal medicines against C. albicans of clinical origin. Accepted Results The Effects of Phenylpropenes and Antifungal Medicines The results of the MICs of tested phenylpropenes and antifungal medicines against C. albicans strains are summarized in Table 1 and Table 2, respectively. The results showed that all yeast strains were susceptible to tested EOCs. The highest inhibiting activity against isolates was observed for eugenol (MIC = 1.96 ± 0.00 mg/mL — 3.92 ± 0.00 mg/mL; MFC = 3.92 ± 0.00 mg/mL — 7.72 ± 0.00 mg/mL). Moreover, only fungicidal power of eugenol against C. albicans strains was exhibited. On the contrary, the lower inhibition for trans-anethole was detected (MIC = 61.75 ± 0.00 mg/mL — 247.00 ± 0.00 mg/mL; MFC = 247.00 ± 0.00 mg/mL — 494.00 ± 0.00 mg/mL). Both the control and clinical isolates were susceptible to tested antifungal medicines. Furthermore, all tested drugs showed fungicidal power against C. albicans strains. The highest and lowest inhibiting activity against isolates were observed for clotrimazole (MIC = 0.01 ± 0.00 mg/mL — 0.02 ± 0.00 mg/mL; MFC = 0.02 ± 0.00 mg/mL — 0.03 ± 0.00 mg/mL), and econazole (MIC = 0.02 ± 0.00 mg/mL — 0.06 ± 0.00 1 Pobrano z https://publicum.umed.lodz.plThis article is protected / Downloaded by copyright. from AllRepository rights reserved. of Medical University of Lodz 2021-10-07 Chemistry & Biodiversity 10.1002/cbdv.202000843 Chem. Biodiversity mg/mL; MFC = 0.04 ± 0.00 mg/mL — 0.13 ± 0.00 mg/mL), respectively. Moreover, it has been also found that the highest inhibiting activity against C. albicans ATCC 10231 strain was to miconazole, econazole and clotrimazole. Table 1. Minimal inhibitory concentration (MIC), minimal fungicidal concentration (MFC), ratio MFC/MIC and power of the essential oil compounds