Skip to main navigation menu Skip to main content Skip to site footer
Plant Science Today (PST)

Research Articles

Vol. 13 No. 3 (2026)

Bioactivity and developmental safety evaluation of Ocimum basilicum L. essential oil using zebrafish model

DOI
https://doi.org/10.14719/pst.12545
Submitted
1 November 2025
Published
15-06-2026 — Updated on 10-08-2026
Versions

Abstract

Ocimum basilicum L. (sweet basil) essential oil (EOOB) is widely used for its antimicrobial and therapeutic benefits; however, its safety and toxicity remain underexplored. This study provides the first integrated investigation of the chemical composition, antibacterial efficacy and developmental toxicity of EOOB using a zebrafish (Danio rerio) model. Ocimum basilicum essential oil (EOOB) was extracted by hydro-distillation and chemically characterised using gas chromatography-mass spectrometry (GC-MS) and Fourier transform infrared spectroscopy (FTIR) spectroscopy. The antibacterial activity of EOOB and its major constituent, methyl cinnamate (MeCin), was evaluated against Bacillus subtilis, Escherichia coli, Salmonella typhi and Staphylococcus aureus using agar well diffusion method and minimum inhibitory concentration (MIC) assays. Embryotoxicity and cardiotoxicity were assessed in zebrafish embryos at 24–96 hours post-fertilisation (hpf) to establish developmental safety profiles. The GC-MS analysis revealed MeCin (≈ 45.6 %) as the major constituent, followed by linalool, geraniol and eucalyptol. Ocimum basilicum essential oil (EOOB) exhibited stronger antibacterial activity than MeCin, suggesting possible synergistic interactions among minor terpenoids. Embryotoxicity assays demonstrated concentration-dependent developmental defects, including pericardial and yolk sac edema, spinal curvature and cardiac arrest, which were more pronounced in MeCin-treated embryos than in EOOB-treated embryos. The zebrafish model enabled direct visualisation of morphological and cardiac effects, providing a robust platform for toxicity assessment. This study is the first to correlate the phytochemical profile of a MeCin-rich O. basilicum chemotype with both antibacterial and embryotoxic outcomes in zebrafish. Overall, EOOB exhibited potent antibacterial activity with relatively lower developmental toxicity than its isolated major constituent, supporting the concept of whole-oil synergy. Furthermore, this study establishes zebrafish-based embryotoxicity screening as a novel, ethical and predictive approach for evaluating the safety of bioactive essential oils.

References

  1. 1. Nabrdalik M, Grata K. Antibacterial activity of Ocimum basilicum L. essential oil against Gram-negative bacteria. Postepy Fitoter. 2016;2:80–6.
  2. 2. Ahmad CM, Naz SB, Sharif A, Akram M, Saeed MA. Biological and pharmacological properties of sweet basil (Ocimum basilicum). Br J Pharm Res. 2015;7:330–9. https://doi.org/10.9734/BJPR/2015/16505
  3. 3. Paton A, Harley RM, Harley MM. Ocimum—an overview of relationships and classification. In: Hiltunen R, Holm Y, editors. Ocimum. Medicinal and aromatic plants – industrial profiles. Amsterdam: Harwood Academic Publishers; 1999. p. 1–38.
  4. 4. Gülçin I, Elmastaş M, Aboul-Enein HY. Determination of antioxidant and radical-scavenging activity of basil (Ocimum basilicum L., Lamiaceae) assayed by different methodologies. Phytother Res. 2007;21:354–61. https://doi.org/10.1002/ptr.2069
  5. 5. Mäkinen SM, Paakkonen KK. Processing and use of basil in foodstuffs, beverages and food preparation. In: Hiltunen R, Holm Y, editors. Basil: the genus Ocimum. Amsterdam: Harwood Academic Publishers; 1999. p. 137–52.
  6. 6. Suppakul P, Miltz J, Sonneveld K, Bigger SW. Antimicrobial properties of basil and its possible application in food packaging. J Agric Food Chem. 2003;51:3197–207. https://doi.org/10.1021/jf021038t
  7. 7. Nour AH, Idris AA, Ishag OA, Nour AH. Chemical composition and repellent activity of methyl cinnamate-rich basil (Ocimum basilicum) essential oil. J Turk Chem Soc A. 2022;9:1277–84. https://doi.org/10.18596/jotcsa.1108807
  8. 8. Joshi RK. Chemical composition and antimicrobial activity of essential oil of Ocimum basilicum L. from the Western Ghats of North-West Karnataka, India. Indian J Nat Prod Resour. 2014;5:151–6. https://doi.org/10.4103/0257-7941.144618
  9. 9. Govindarajan M, Sivakumar R, Rajeswary M, Yogalakshmi K. Chemical composition and larvicidal activity of essential oil from Ocimum basilicum (L.) against mosquito vectors. Exp Parasitol. 2013;134:7–11. https://doi.org/10.1016/j.exppara.2013.01.018
  10. 10. Hussain AI, Anwar F, Sherazi STH, Przybylski R. Chemical composition, antioxidant and antimicrobial activities of basil (Ocimum basilicum) essential oils depending on seasonal variations. Food Chem. 2008;108:986–95. https://doi.org/10.1016/j.foodchem.2007.12.010
  11. 11. Kathirvel R, Ravi S. Chemical composition of basil (Ocimum basilicum L.) essential oil and its in vitro cytotoxicity against human cancer cell lines. Nat Prod Res. 2012;26:1112–8. https://doi.org/10.1080/14786419.2010.545357
  12. 12. Özcan M, Chalchat JC. Essential oil composition of Ocimum basilicum L. and Ocimum minimum L. in Turkey. Czech J Food Sci. 2002;20:223–8. https://doi.org/10.17221/3536-CJFS
  13. 13. Politeo O, Jukić M, Miloš M. Chemical composition and antioxidant capacity of free volatile aglycones from basil (Ocimum basilicum L.). Food Chem. 2007;101:379–85. https://doi.org/10.1016/j.foodchem.2006.01.045
  14. 14. Saha S, Dhar TN, Sengupta C, Ghosh P. Biological activities of essential oils and methanol extracts of five Ocimum species. Czech J Food Sci. 2013;31:194–202. https://doi.org/10.17221/234/2012-CJFS
  15. 15. Shirazi MT, Gholami H, Kavoosi G, Rowshan V, Tafsiry A. Chemical composition, antioxidant, antimicrobial and cytotoxic activities of Tagetes minuta and Ocimum basilicum essential oils. Food Sci Nutr. 2014;2:146–55. https://doi.org/10.1002/fsn3.85
  16. 16. Verma RS, Bisht PS, Padalia RC, Saikia D, Chauhan A. Chemical composition and antibacterial activity of essential oil from two Ocimum species grown in India. Asian J Tradit Med. 2011;6:211–7.
  17. 17. Beatović D, Krstić-Milošević D, Trifunović S, Šiljegović J, Glamočlija J, Ristić M, et al. Chemical composition, antioxidant and antimicrobial activities of essential oils of twelve Ocimum basilicum L. cultivars grown in Serbia. Rec Nat Prod. 2015;9:62–75.
  18. 18. Romagosa CMR, David ES, Dulay RMR. Embryotoxic and teratogenic effects of Tinospora cordifolia extracts in zebrafish embryos. Asian J Plant Sci Res. 2016;6:37–41.
  19. 19. Ng’uni T, Klaasen JA, Fielding BC. Acute toxicity studies of the South African medicinal plant Galenia africana. Toxicol Rep. 2018;5:813–8. https://doi.org/10.1016/j.toxrep.2018.08.008
  20. 20. Burt S. Essential oils: antibacterial properties and applications in foods. Int J Food Microbiol. 2004;94:223–53. https://doi.org/10.1016/j.ijfoodmicro.2004.03.022
  21. 21. Alafiatayo A, Lai KS, Syahida A, Mahmood M, Shaharuddin N. Phytochemical evaluation and embryotoxicity of Curcuma longa extract on zebrafish (Danio rerio). Evid Based Complement Alternat Med. 2019;2019(1):3807207. https://doi.org/10.1155/2019/3807207
  22. 22. Ali M, Saba S, Taite D, Emadi S, Irving R. Protective layer changes in zebrafish embryos with age. J Toxicol Pharmacol. 2017;1:1–8.
  23. 23. Cassar S, et al. Use of zebrafish in drug discovery toxicology. Chem Res Toxicol. 2020;33:95–118. https://doi.org/10.1021/acs.chemrestox.9b00335
  24. 24. Ducharme NA, Reif DM, Gustafsson JA, Bondesson M. Comparison of zebrafish and mammalian toxicity values. Reprod Toxicol. 2015;55:3–10. https://doi.org/10.1016/j.reprotox.2014.09.005
  25. 25. Bassolé IHN, Lamien-Meda A, Bayala B, Tirogo S, Franz C, Novak J, et al. Composition and antimicrobial activities of essential oils. Molecules. 2010;15:7825–39. https://doi.org/10.3390/molecules15117825
  26. 26. Hudzicki J. Kirby-Bauer disk diffusion susceptibility test protocol. Washington (DC): American Society for Microbiology; 2009.
  27. 27. Ashraf SA, Al Shammari E, Hussain T, Tajuddin S, Panda BP. In vitro antimicrobial activity and GC–MS identification of essential oils. J Food Sci Technol. 2017;54:3948–58. https://doi.org/10.1007/s13197-017-2859-2
  28. 28. Perumal S, Pillai S, Cai LW, Mahmud R, Ramanathan S. Determination of minimum inhibitory concentration using tetrazolium microplate assay. J Nat Prod. 2012;5:68–76.
  29. 29. Kimmel CB, Ballard WW, Kimmel SR, Ullmann B, Schilling TF. Stages of embryonic development of the zebrafish. Dev Dyn. 1995;203:253–310. https://doi.org/10.1002/aja.1002030302
  30. 30. De Luca E, et al. ZebraBeat: a platform for analysis of cardiac rate in zebrafish embryos. Sci Rep. 2014;4:4898. https://doi.org/10.1038/srep04898
  31. 31. Kumar RBS, Kar B, Dolai N, Haldar PK. Developmental toxicity of Streblus asper bark in zebrafish embryos. Indian J Nat Prod Resour. 2013;4:255–9.
  32. 32. Khan BA, Ullah S, Khan MK, Alshahrani SM, Braga VA. Formulation and evaluation of Ocimum basilicum-based emulgel for wound healing. Saudi Pharm J. 2020;28:1842–50. https://doi.org/10.1016/j.jsps.2020.11.011
  33. 33. Colthup NB, Daly LH, Wiberley SE. Introduction to infrared and Raman spectroscopy. 3rd ed. Massachusetts: Academic Press; 1990.
  34. 34. Wade LG. Organic chemistry. LibreTexts Project; 2026.
  35. 35. Deviani AE, Widan A, Hanhadyanaputri ES. Synthesis of n-octyl cinnamate and antioxidant activity. Med Sains J Ilm Kefarm. 2025;10:1. https://doi.org/10.37874/ms.v10i2.1714
  36. 36. Alakomi HL, Skyttä E, Saarela M, Mattila-Sandholm T, Latva-Kala K, Helander IM. Lactic acid permeabilizes Gram-negative bacteria. Appl Environ Microbiol. 2000;66:2001–5. https://doi.org/10.1128/AEM.66.5.2001-2005.2000
  37. 37. Pagès JM, James CE, Winterhalter M. Porins and antibiotic permeation in Gram-negative bacteria. Nat Rev Microbiol. 2008;6:893–903. https://doi.org/10.1038/nrmicro1994
  38. 38. Smith-Palmer A, Stewart J, Fyfe L. Antimicrobial properties of plant essential oils. Lett Appl Microbiol. 1998;26:118–22. https://doi.org/10.1046/j.1472-765X.1998.00303.x
  39. 39. Eriotou E, Anastasiadou K, Nikolopoulos D, Koulougliotis D. Antimicrobial and antioxidant activities of basil essential oil. J Nutr Food Sci. 2016;5:1–9.
  40. 40. Veras HNH, Rodrigues FFG, Colares AV, Menezes IRA, Coutinho HDM, Botelho MA, et al. Synergistic antibiotic activity of essential oils. Fitoterapia. 2012;83:508–12. https://doi.org/10.1016/j.fitote.2011.12.024
  41. 41. Paparella A, Lorenzo Taccogna L, Aguzzi I, Chaves-López C, Serio A, Marsilio F, et al. Flow cytometric assessment of antimicrobial activity of essential oils. Food Control. 1999;9:1174–82. https://doi.org/10.1016/j.foodcont.2008.01.002
  42. 42. Xu J, Zhou F, Ji BP, Pei RS, Xu N. Antibacterial mechanism of carvacrol and thymol. Lett Appl Microbiol. 2008;47:174–9. https://doi.org/10.1111/j.1472-765X.2008.02407.x
  43. 43. Kuorwel KK, Cran MJ, Sonneveld K, Miltz J, Bigger SW. Essential oils as antimicrobial agents for packaging films. J Food Sci. 2011;76:R164–77. https://doi.org/10.1111/j.1750-3841.2011.02384.x
  44. 44. Ultee A, Kets EP, Smid EJ. Mechanisms of action of carvacrol. Appl Environ Microbiol. 1999;65:4606–10. https://doi.org/10.1128/AEM.65.10.4606-4610.1999
  45. 45. Juven BJ, Kanner J, Schved F, Weisslowicz H. Factors interacting with thyme oil antibacterial action. J Appl Bacteriol. 1994;76:626–31. https://doi.org/10.1111/j.1365-2672.1994.tb01661.x
  46. 46. Damiani E, Sella F, Astolfi P, Galeazzi R, Carnevali O, Maradonna F. Effects of Tempol-Methoxycinnamate on zebrafish development. Int J Mol Sci. 2023;24:6767. https://doi.org/10.3390/ijms24076767

Downloads

Download data is not yet available.