Antibacterial Activity against Staphylococcus aureus and In Vitro Cytotoxicity of a Pimpinella anisum Essential Oil Nanoemulsion
DOI:
https://doi.org/10.15649/2346075X.6281Keywords:
Essential oil, Nanoemulsion, Pimpinella anisum, citotoxicity, Staphylococcus aureus, Antimicrobial activityAbstract
Introduction. Microbial resistance to disinfectants is an increasing challenge because it can promote pathogen persistence on surfaces and devices. Staphylococcus aureus is characterized by its ability to form biofilms and cause healthcare-associated infections and contamination in food-related environments. In this context, essential oils such as that of Pimpinella anisum L. and their nanoemulsions have attracted interest because of their potential antimicrobial properties. Objective. To evaluate the antimicrobial activity and cytotoxicity of a Pimpinella anisum essential oil nanoemulsion against Staphylococcus aureus and Vero cells. Materials and Methods. A P. anisum nanoemulsion (NEPa) was prepared and colloidally characterized. Cytotoxicity was assessed in Vero cells using the MTT colorimetric assay. Antimicrobial activity against S. aureus was evaluated by a direct-contact method using concentrations of 300 and 600 mg/L and exposure times of 1, 3, 5, and 10 min. Results. A monodisperse NEPa with a nanoscale hydrodynamic diameter was obtained. The nanoemulsion showed a CC50 of 185.3 mg/L and a CC90 above 740.74 mg/L. At 600 mg/L, NEPa reduced S. aureus growth by 82.3% after 1 min and by 86.4% after 10 min of exposure. Conclusions. P. anisum NEPa showed antimicrobial activity against S. aureus and concentration-dependent cytotoxicity. These findings support its potential as an antimicrobial system; however, further studies are required to optimize concentration and exposure conditions and to establish an appropriate balance between antimicrobial efficacy and cytotoxicity.
References
1. Rozman U, Pušnik M, Kmetec S, Duh D, Šostar Turk S. Reduced susceptibility and increased resistance of bacteria against disinfectants: a systematic review. Microorganisms. 2021;9(12):2550. https://doi.org/10.3390/microorganisms9122550
2. Walczak ŁJ, Kwiatkowska M, Twarowski B, Kubacka M, Paluch J, Herbet M. Disinfectant-induced bacterial resistance and antibiotic cross-resistance—mechanisms and clinical relevance. Clin Exp Med. 2026;26:26. https://doi.org/10.1007/s10238-025-01950-2
3. De Oliveira DMP, Forde BM, Kidd TJ, Harris PNA, Schembri MA, Beatson SA, et al. Antimicrobial resistance in ESKAPE pathogens. Clin Microbiol Rev. 2020;33(3):e00181-19. https://doi.org/10.1128/CMR.00181-19
4. Pasachova Garzón J, Ramirez Martinez S, Muñoz Molina L. Staphylococcus aureus: generalidades, mecanismos de patogenicidad y colonización celular. Nova. 2019;17(32):25-38. https://doi.org/10.22490/24629448.3631
5. Li Q, Liu L, Guo A, Zhang X, Liu W, Ruan Y. Formation of multispecies biofilms and their resistance to disinfectants in food processing environments: a review. J Food Prot. 2021;84(12):2071-2083. https://doi.org/10.4315/JFP-21-071
6. Sun W, Shahrajabian MH, Cheng Q. Anise (Pimpinella anisum L.), a dominant spice and traditional medicinal herb for both food and medicinal purposes. Cogent Biol. 2019;5(1):1673688. https://doi.org/10.1080/23312025.2019.1673688
7. Pino O, Sánchez Y, Rojas MM, Abreu Y, Correa TM. Composición química y actividad antibacteriana del aceite esencial de Pimpinella anisum L. Rev Protección Veg. 2012;27(3):181-187.
8. Dumitrescu E, Muselin F, Tîrziu E, Folescu M, Dumitrescu CS, Orboi DM, et al. Pimpinella anisum L. essential oil: a valuable antibacterial and antifungal alternative. Plants (Basel). 2023;12(13):2428. https://doi.org/10.3390/plants12132428
9. Pavoni L, Perinelli DR, Bonacucina G, Cespi M, Palmieri GF. An overview of micro- and nanoemulsions as vehicles for essential oils: formulation, preparation and stability. Nanomaterials (Basel). 2020;10(1):135. https://doi.org/10.3390/nano10010135
10. Posit team. RStudio: integrated development environment for R [Internet]. Boston (MA): Posit Software, PBC; 2024. Available from: https://posit.co
11. Omidian H, Cubeddu LX, Gill EJ. Harnessing nanotechnology to enhance essential oil applications. Molecules. 2025;30(3):520. https://doi.org/10.3390/molecules30030520
12. Malode GP, Ande SN, Chavhan SA, Bartare SA, Malode LL, Manwar JV, Bakal RL. A critical review on nanoemulsion: advantages, techniques and characterization. World J Adv Res Rev. 2021;11(3):462-473. https://doi.org/10.30574/wjarr.2021.11.3.0431
13. Dadkhah F, Aliakbarlu J, Tajik H. Determining optimum conditions for biofilm formation by Salmonella isolates and anti-biofilm activity of cinnamon essential oil nanoemulsion. Biofouling. 2025;41(5):485-497. https://doi.org/10.1080/08927014.2025.2499714
14. Da Silva B, Rosário DD, Neto L, Lelis C, Conte-Junior C. Antioxidant, antibacterial and antibiofilm activity of nanoemulsion-based natural compound delivery systems compared with non-nanoemulsified versions. Foods. 2023;12(9):1901. https://doi.org/10.3390/foods12091901
15. Zhao D, Ma Y, Wang W, Xiang Q. Antibacterial activity and mechanism of cinnamon essential oil nanoemulsion against Pseudomonas deceptionensis CM2. Heliyon. 2023;9(9):e19582. https://doi.org/10.1016/j.heliyon.2023.e19582
16. Shehabeldine AM, Doghish AS, El-Dakroury WA, Hassanin MMH, Al-Askar AA, AbdElgawad H, et al. Antimicrobial, antibiofilm, and anticancer activities of Syzygium aromaticum essential oil nanoemulsion. Molecules. 2023;28(15):5812. https://doi.org/10.3390/molecules28155812
17. Donsì F, Annunziata M, Sessa M, Ferrari G. Nanoencapsulation of essential oils to enhance their antimicrobial activity in foods. LWT Food Sci Technol. 2011;44(9):1908-1914. https://doi.org/10.1016/j.lwt.2011.03.003
18. Gupta A, Eral HB, Hatton TA, Doyle PS. Nanoemulsions: formation, properties and applications. Soft Matter. 2016;12:2826-2841. https://doi.org/10.1039/C5SM02958A
19. Choi SJ, McClements DJ. Nanoemulsions as delivery systems for lipophilic nutraceuticals: strategies for improving their formulation, stability, functionality and bioavailability. Food Sci Biotechnol. 2020;29(2):149-168. https://doi.org/10.1007/s10068-019-00731-4
20. Hashem AS, Awadalla SS, Zayed GM, Maggi F, Benelli G. Pimpinella anisum essential oil nanoemulsions against Tribolium castaneum: insecticidal activity and mode of action. Environ Sci Pollut Res Int. 2018;25(19):18802-18812. https://doi.org/10.1007/s11356-018-2068-1
21. Azeem A, Rizwan M, Ahmad FJ, Khar RK, Iqbal Z, Talegaonkar S. Components screening and influence of surfactant and cosurfactant on nanoemulsion formation. Curr Nanosci. 2009;5(2):220-226. https://doi.org/10.2174/157341309788185505
22. Sun H, Luo D, Zheng S, Li Z, Xu W. Antimicrobial behavior and mechanism of clove oil nanoemulsion. J Food Sci Technol. 2022;59(5):1939-1947. https://doi.org/10.1007/s13197-021-05208-z
23. Hashem AS, Ramadan MM, Abdel-Hady AAA, Sut S, Maggi F, Dall’Acqua S. Pimpinella anisum essential oil nanoemulsion toxicity against Tribolium castaneum? Shedding light on its interactions with aspartate aminotransferase and alanine aminotransferase by molecular docking. Molecules. 2020;25(20):4841. https://doi.org/10.3390/molecules25204841
24. Abu Ali OA, El-Naggar ME, Abdel-Aziz MS, Saleh DI, Abu-Saied MA, El-Sayed WA. Facile synthesis of natural anise-based nanoemulsions and their antimicrobial activity. Polymers (Basel). 2021;13(12):2009. https://doi.org/10.3390/polym13122009
25. McClements DJ. Nanoemulsions versus microemulsions: terminology, differences, and similarities. Soft Matter. 2012;8:1719-1729. https://doi.org/10.1039/C2SM06903B
26. Brito GS, Dutra RP, Pereira ALF, Ferreira AGN, Santos Neto M, Holanda CA, Fidelis QC. Nanoemulsions of essential oils against multi-resistant microorganisms: an integrative review. Microb Pathog. 2024;195:106837. https://doi.org/10.1016/j.micpath.2024.106837
27. Aswathanarayan JB, Vittal RR. Nanoemulsions and their potential applications in food industry. Front Sustain Food Syst. 2019;3:95. https://doi.org/10.3389/fsufs.2019.00095
28. Costa SS, Viveiros M, Rosato AE, Melo-Cristino J, Couto I. Impact of efflux in the development of multidrug resistance phenotypes in Staphylococcus aureus. BMC Microbiol. 2015;15:232. https://doi.org/10.1186/s12866-015-0572-8
29. Singh Y, Meher JG, Raval K, Khan FA, Chaurasia M, Jain NK, Chourasia MK. Nanoemulsion: concepts, development and applications in drug delivery. J Control Release. 2017;252:28-49. https://doi.org/10.1016/j.jconrel.2017.03.008
30. Jerobin J, Makwana P, Suresh Kumar RS, Sundaramoorthy R, Mukherjee A, Chandrasekaran N. Antibacterial activity of neem nanoemulsion and its toxicity assessment on human lymphocytes in vitro. Int J Nanomedicine. 2015;10 Suppl 1:77-86. https://doi.org/10.2147/IJN.S79983
31. Rahamooz-Haghighi S, Asadi MH. Anti-proliferative effect of the extracts and essential oil of Pimpinella anisum on gastric cancer cells. J Herbmed Pharmacol. 2016;5(4):157-161. Sin DOI confirmado.
32. Nagaraju PG, Sengupta P, Chicgovinda PP, Rao PJ. Nanoencapsulation of clove oil and study of physicochemical properties, cytotoxic, hemolytic, and antioxidant activities. J Food Process Eng. 2021;44(4):e13645. https://doi.org/10.1111/jfpe.13645
33. Aguilera D, Hidrobo A, Venegas J. Toxicidad del hipoclorito de sodio y compuestos de amonio cuaternario en seres humanos. Una revisión sistemática en época de COVID-19. Rev Toxicol. 2023;40(1):53-66. Sin DOI confirmado.
34. Abd-Elhafeez EMA, Ramadan BR, Abou-El-Hawa SHM, Rashwan MRA. Chemical composition and antimicrobial activity of anise and fennel essential oils. Assiut J Agric Sci. 2023;54(2):127-140. https://doi.org/10.21608/ajas.2023.177714.1211
35. Contant C, Rouabhia M, Loubaki L, Chandad F, Semlali A. Anethole induces anti-oral cancer activity by triggering apoptosis, autophagy and oxidative stress and by modulation of multiple signaling pathways. Sci Rep. 2021;11(1):13087. https://doi.org/10.1038/s41598-021-92456-w
36. Fujita KI, Tatsumi M, Ogita A, Kubo I, Tanaka T. Anethole induces apoptotic cell death accompanied by reactive oxygen species production and DNA fragmentation in Aspergillus fumigatus and Saccharomyces cerevisiae. FEBS J. 2014;281(4):1304-1313. https://doi.org/10.1111/febs.12706
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