Natural Preservation of Horticultural Produce: Antimicrobial Efficacy of Thymus hiemalis Essential Oil in in Vitro and in Situ Models

PDF

Authors: Joel H. Elizondo-Luevano, Guadalupe Gutiérrez-Soto, Iosvany López-Sandin, Georgia M. González-Meza, Lucio Galaviz-Silva, Catalina Leos-Rivas, Osvelia E. Rodríguez-Luis and Julio López-Abán

Volume/Issue: Volume 28: Issue 2

Published online: 18 Nov 2025

Pages: 150 - 158

DOI: https://doi.org/10.2478/ahr-2025-0019


Abstract

The present study evaluated the antimicrobial activity of Thymus hiemalis essential oil (THEO) against selected phytopathogenic bacteria and fungi using both in vitro and in situ approaches. Seven plant pathogens were tested: Xanthomonas arboricola, Pectobacterium carotovorum, Pseudomonas syringae, Agrobacterium radiobacter, Fusarium solani, Monilia fructigena, and Botrytis cinerea. In vitro antimicrobial activity was assessed using the disk diffusion method and determination of minimum inhibitory concentrations (MICs). THEO exhibited broad-spectrum antimicrobial effects, with the strongest inhibition observed in fungal isolates, particularly B. cinerea and F. solani. In situ assays were conducted on fresh-cut horticultural produce models – strawberry, apple, carrot, and parsley – to simulate natural microbial contamination. The antimicrobial efficacy of THEO in the vapour phase was tested at concentrations of 500, 250, 125, and 62.5 µL.L−1. Growth inhibition was quantified using stereological image analysis, and the percentage reduction in microbial volume density (Vv) was calculated. Statistically significant inhibitory effects were observed across all tested matrices, especially at higher concentrations. The highest biological growth inhibition (BGI) values were recorded for F. solani and M. fructigena, indicating strong fungistatic potential of THEO vapours in horticultural systems. These findings suggest that Thymus hiemalis essential oil possesses effective antimicrobial properties with practical relevance for horticultural applications, particularly in the control of postharvest spoilage and decay caused by phytopathogens.


Keywords: Thymus hiemalis essential oil, antimicrobial activity, phytopathogens, in situ analysis, postharvest decay, horticultural produce, minimum inhibitory concentration, stereology

PDF

References

Ben Miri, Y. (2025). Essential Oils: Chemical Composition and Diverse Biological Activities : A Comprehensive Review. Natural Product Communications, 20(1), 1934578X241311790. https://doi.org/10.1177/1934578X241311790


Bisht, A., & Singh, S. P. (2024). Postharvest Losses and Management of Horticultural Produce: A Review. Journal of Scientific Research and Reports, 30(3), 305–320. https://doi.org/10.9734/jsrr/2024/v30i31881


Bozin, B., Mimica-Dukic, N., Simin, N., & Anackov, G. (2006). Characterization of the Volatile Composition of Essential Oils of Some Lamiaceae Spices and the Antimicrobial and Antioxidant Activities of the Entire Oils. Journal of Agricultural and Food Chemistry, 54(5), 1822–1828. https://doi.org/10.1021/jf051922u


Bukovská, A., Cikoš, Š., Juhás, Š., Il’ková, G., Rehák, P., & Koppel, J. (2007). Effects of a Combination of Thyme and Oregano Essential Oils on TNBS-Induced Colitis in Mice. Mediators of Inflammation, 2007, 1–9. https://doi.org/10.1155/2007/23296


Burt, S. (2004). Essential oils: Their antibacterial properties and potential applications in foods – a review. International Journal of Food Microbiology, 94(3), 223–253. https://doi.org/10.1016/j.ijfoodmicro.2004.03.022


De Martino, L., Bruno, M., Formisano, C., De Feo, V., Napolitano, F., Rosselli, S., & Senatore, F. (2009). Chemical Composition and Antimicrobial Activity of the Essential Oils from Two Species of Thymus Growing Wild in Southern Italy. Molecules, 14(11), 4614–4624. https://doi.org/10.3390/molecules14114614


Dorman, H. J. D., & Deans, S. G. (2000). Antimicrobial agents from plants: Antibacterial activity of plant volatile oils. Journal of Applied Microbiology, 88(2), 308–316. https://doi.org/10.1046/j.1365-2672.2000.00969.x


Etri, K., & Pluhár, Z. (2024). Exploring Chemical Variability in the Essential Oils of the Thymus Genus. Plants, 13(10), 1375. https://doi.org/10.3390/plants13101375


Fenta, L., & Mekonnen, H. (2024). Microbial Biofungicides as a Substitute for Chemical Fungicides in the Control of Phytopathogens: Current Perspectives and Research Directions. Scientifica, 2024, 1–12. https://doi.org/10.1155/2024/5322696


Hossain, T. J. (2024). Methods for screening and evaluation of antimicrobial activity: A review of protocols, advantages, and limitations. European Journal of Microbiology and Immunology, 14(2), 97–115. https://doi.org/10.1556/1886.2024.00035


Kačániová, M., Galovičová, L., Valková, V., Ďuranová, H., Borotová, P., Štefániková, J., Vukovic, N. L., Vukic, M., Kunová, S., Felsöciová, S., Miklášová, K., Savitskaya, T., & Grinshpan, D. (2021). Chemical composition and biological activity of Salvia officinalis essential oil. Acta Horticulturae et Regiotecturae, 24(2), 81–88. https://doi.org/10.2478/ahr-2021-0028


Kluz, M. I., & Vukovic, N. (2025). Antimicrobial Activity of Jasminum grandiflorum Absolute in vitro and in situ Study against Phytopathogenic Bacteria. Acta Horticulturae et Regiotecturae, 28(1), 66–71. https://doi.org/10.2478/ahr-2025-0008


Laird, K., & Phillips, C. (2012). Vapour phase: A potential future use for essential oils as antimicrobials? Essential oil vapours and their antimicrobial activity. Letters in Applied Microbiology, 54(3), 169–174. https://doi.org/10.1111/j.1472-765X.2011.03190.x


Lopez-Reyes, J. G., Spadaro, D., Prelle, A., Garibaldi, A., & Gullino, M. L. (2013). Efficacy of Plant Essential Oils on Postharvest Control of Rots Caused by Fungi on Different Stone Fruits In Vivo. Journal of Food Protection, 76(4), 631–639. https://doi.org/10.4315/0362-028X.JFP-12-342


Nazzaro, F., Fratianni, F., Coppola, R., & Feo, V. D. (2017). Essential Oils and Antifungal Activity. Pharmaceuticals, 10(4), 86. https://doi.org/10.3390/ph10040086


Opara, U. L., & Ogra, I. O. (2024). An Introduction to Postharvest Handling Technology of Fresh Fruits and Vegetables. S. Ali, S. A. Mir, B. N. Dar, & S. Ejaz. Sustainable Postharvest Technologies for Fruits and Vegetables (1st ed., pp. 3–41). https://doi.org/10.1201/9781003370376-2


Passone, M. A., Girardi, N. S., Ferrand, C. A., & Etcheverry, M. (2012). In vitro evaluation of five essential oils as botanical fungitoxicants for the protection of stored peanuts from Aspergillus flavus and A. parasiticus contamination. International Biodeterioration & Biodegradation, 70, 82–88. https://doi.org/10.1016/j.ibiod.2011.11.017


Reyes-Jurado, F., Navarro-Cruz, A. R., Ochoa-Velasco, C. E., Palou, E., López-Malo, A., & Ávila-Sosa, R. (2020). Essential oils in vapor phase as alternative antimicrobials: A review. Critical Reviews in Food Science and Nutrition, 60(10), 1641–1650. https://doi.org/10.1080/10408398.2019.1586641


Rios, J. L., Recio, M. C., & Villar, A. (1988). Screening methods for natural products with antimicrobial activity: A review of the literature. Journal of Ethnopharmacology, 23(2–3), 127–149. https://doi.org/10.1016/0378-8741(88)90001-3


Rota, M. C., Herrera, A., Martínez, R. M., Sotomayor, J. A., & Jordán, M. J. (2008). Antimicrobial activity and chemical composition of Thymus vulgaris, Thymus zygis and Thymus hyemalis essential oils. Food Control, 19(7), 681–687. https://doi.org/10.1016/j.foodcont.2007.07.007


Salehi, B., Upadhyay, S., Erdogan Orhan, I., Kumar Jugran, A., L.D. Jayaweera, S., A. Dias, D., Sharopov, F., Taheri, Y., Martins, N., Baghalpour, N., C. Cho, W., & Sharifi-Rad, J. (2019). Therapeutic Potential of α- and β-Pinene: A Miracle Gift of Nature. Biomolecules, 9(11), 738. https://doi.org/10.3390/biom9110738


Semeniuc, C. A., Pop, C. R., & Rotar, A. M. (2017). Antibacterial activity and interactions of plant essential oil combinations against Gram-positive and Gram-negative bacteria. Journal of Food and Drug Analysis, 25(2), 403–408. https://doi.org/10.1016/j.jfda.2016.06.002


Sharma, A., Rajendran, S., Srivastava, A., Sharma, S., & Kundu, B. (2017). Antifungal activities of selected essential oils against Fusarium oxysporum f. Sp. Lycopersici 1322, with emphasis on Syzygium aromaticum essential oil. Journal of Bioscience and Bioengineering, 123(3), 308–313. https://doi.org/10.1016/j.biosc.2016.09.011


Sokolić-Mihalak, D., Frece, J., Slavica, A., Delaš, F., Pavlović, H., & Markov, K. (2012). The Effects Of Wild Thyme (Thymus serpyllum L.) Essential Oil Components Against Ochratoxin-Producing Aspergilli/Majčina Dušica (Thymus serpyllum L.) I Njezine Komponente Protiv Okratoksikotvornih Vrsta Aspergillusa. Archives of Industrial Hygiene and Toxicology, 63(4), 457–462. https://doi.org/10.2478/10004-1254-63-2012-2309


Tzortzakis, N. G., & Economakis, C. D. (2007). Antifungal activity of lemongrass (Cympopogon citratus L.) essential oil against key postharvest pathogens. Innovative Food Science & Emerging Technologies, 8(2), 253–258. https://doi.org/10.1016/j.ifset.2007.01.002


Zubair, M., Shahzad, S., Hussain, A., Pradhan, R. A., Arshad, M., & Ullah, A. (2022). Current Trends in the Utilization of Essential Oils for Polysaccharide- and Protein-Derived Food Packaging Materials. Polymers, 14(6), 1146. https://doi.org/10.3390/polym14061146