• Dalia Makkia Animal Health Research Institute, Mansoura Provincial Laboratory, Agriculture Research Centre, Ministry of Agriculture, Egypt
  • Ali Bahout Food Control Department, Faculty of Veterinary Medicine, Zagazig University, Egypt
  • Mohamed A. Bayoumi Food Control Department, Faculty of Veterinary Medicine, Zagazig University, Egypt
  • Mohamed E. Alnakip Food Control Department, Faculty of Veterinary Medicine, Zagazig University, Egypt, Corresponding author, E-mail: alnakip.me@gmail.com
  • Adel H. Moustafa Animal Health Research Institute, Mansoura Provincial Laboratory, Agriculture Research Centre, Ministry of Agriculture, Egypt




essential oils, thyme oil, black seed oil, cheese, E. coli, A. hydrophila


Obtaining healthy food, free from chemical or synthetic additives, is a major challenge. In this study, we developed a preservation method using essential oils and evaluated their effect on multidrug resistant pathogenic Gram-negative bacteria. Different concentrations (1%,0.1%,0.17% and 0.35%) of Thyme oil and black seed oil were employed in this study against pathogenic E. coli and A. hydrophila in soft cheese. The used oils at a concentration of 0.1% through dipping method resulted in accepted color and odor, little effect on flavor and normal texture and appearance, while 0.1% during inoculation had the same effect as in dipping method except moderate odor. Thyme oil had the highest reduction rate in case of 1% dipping and 0.17% inoculation on A. hydrophila, while against E. coli it was found that 0.17% inoculation and 0.35% of the dipping method had the highest reduction effect. Thyme essential oil seems to be a suitable natural food preservative alternative.


● 1. WHO .World Health Organization Report. Food safety and foodborne illness. World Health Organization Fact sheet 237, 2002; Geneva.

● 2. Mohamed S H S , Zaky WM, Kassem JM, Abbas HM, Salem, M M E, Said-Al-Ahl HAH. Impact of antimicrobial properties of some essential oils on cheese yoghurt quality. World Appl Sci J 2013; 27 (4): 497–507.

● 3. Elhadidy M, Mohammed MA. Shiga toxin-producing Escherichia coli from raw milk cheese in Egypt: Prevalence, molecular characterization and survival to stress conditions. Lett Appl Microbiol 2013; 56 (2): 120–7.

● 4. Lara VM, Carregaro AB, Santurio DF, de Sa MF, Santurio JM, Alves SH. Antimicrobial susceptibility of Escherichia coli strains isolated from Alouatta spp. feces to essential oils. Evid Based Compl Altern Med 2016; 10: 30.

● 5. Eid SAS , Erfan AM.Characterization of E. coli associated with high mortality of poultry flocks. Assiut Vet Med J 2013; 59 (139): 51–61, Egypt.

● 6. Igbinosa I H, Igumbor E U, Aghdasi F, Tom M, Okoh A I. Emerging Aeromonas species infections and their significance in public health. The Scientific Wrld J 2012; 625023, 1–13. doi:10.1100/2012/625023

● 7. Aminzare M, Hashemi M, Hassanzad Azar H, Hejazi J. The Use of Herbal Extracts and Essential Oils as a Potential Antimicrobial in Meat and Meat Products; A Review. J of Human, Environment, and Health Promotion. 2016; 1 (2): 63–74.

● 8. Moro A, Librán CM, Berruga MI, Carmona M, Zalacain A.Dairy matrix effect on the transference of rosemary essential oil compounds during cheese making. J of the Science of Food and Agriculture 2015; 95: 1507–13.

● 9. Es I, Khaneghah A M , Akbariirad H. Global regulation of essential oils. Essential oils in food processing: chemistry, safety and applications. 2017; 327–38.

● 10. Bakhtiary F, Sayevand H R, Khaneghah A M, Haslberger A G, Hosseini H. Antibacterial efficacy of essential oils and sodium nitrite in vacuum processed beef fillet. Applied Food Biotechnology, 2018; 5 (1): 1–10.

● 11. Ismail AM, Harby S , Salem AS. Pro¬duction of flavored labneh with extended shelf life. Egyptian J of Dairy Science 2006; 34(1): 59–68.

● 12. Al-Nabulsi A A, Awaisheh S S, Osaili T M, Olaimat A N, Rahahaleh R J, Al-Dabbas F M, Al-Kharabsheh L A, Gyawali R, Ibrahim S A. Inactivation of Cronobacter sakazakii in reconstituted infant milk formula by plant essential oils. J of Applied Botany and Food Quality 2015; 88 (1): 97–101.

● 13. Amirdivani S , Baba A S. Changes in yogurt fermentation characteristics, and antioxidant potential and in vitro inhibition of angiotensin – 1 converting enzyme upon the inclusion of peppermint, dill and basil. LWT – Food Sci Technol 2011; 44 (6): 1458–64.

● 14. Vinceković M, Viskić M, Jurić S, Giacometti J, Bursać Kovačević D, Putnik P, Režek Jambrak A. Innovative technologies for encapsulation of Mediterranean plants extracts. Trends in Food Science and Technology 2017; 69: 1–12.

● 15. Shen Z, Kamdem D P. Development and characterization of biodegradable chitosan films containing two essential oils. Int J of Biological Macromolecules. 2015; 74: 289–96.

● 16. Mlian KS, Hafiz RS, Muhammad N. Sensory evaluation of consumers acceptability. Hand Book of Food Science and Technology, 2017; 362–86.

● 17. Zarzuelo A, Crespo E. Medicinal and Aromatic Plants–Industrial Profiles, vol. 24. Thyme, the genus Thymus. Stahl-Biskup, E and Sáez, F (Eds.). Taylor and Francis, New York. 2002; 278–92.

● 18. Gutierrez J, Barry-Ryan C, Bourke, P. Antimicrobial activity of plant essential oils using food model media: efûcacy, synergistic potential and interaction with food components. Food Microbiol 2009; 26: 142–50.

● 19. Stahl-Biskup E, Saez, F. Thyme, the Genus Thymus. Taylor and Francis, London, 2002; 331.

● 20. Burt S. Essential oils: their antibacterial properties and potential applications in foods—a review. Int J Food Microbiol 2004; 94: 223–53.

● 21. Shan B, Cai Y Z, Brooks J D ,Corke H. Potential application of spice and herb extracts as natural preservatives in cheese. J of Medicinal Food 2011; 14: 284–90.

● 22. Dagdelen S, Bilenler T, Durmaz G, Gokbulut I, Hayaloglu AA, Karabulut I.Volatile composition, antioxidant and antimicrobial activities of herbal plants used in the manufacture of van herby (OTLU) cheese. J of Food Processing and Preservation, 2014; 38 (4): 1716–25 .

● 23. Salman MT, Khan RA , Shukla I. A study of Nigella sativa seed for antimicrobial activity with special reference to resistant bacteria. 2005;http://openmed.nic.in/49/01/9- Summary.doc.

● 24. Hassoun A , Çoban Ö E .Essential oils for antimicrobial and antioxidant applications in fish and other seafood products. Trends in Food Science and Technology 2017; 68: 26–36.

● 25. Zhang Y. Liu X, Wang Y, Jiang P, Quek S.Antibacterial activity and mechanism of cinnamon essential oil against Escherichia coli and Staphylococcus aureus. Food Control 2016; 59: 282–9.

● 26. Gill AO, Holley RA. Inhibition of membrane bound ATPases of Escherichia coli and Listeria monocytogenes by plant oil aromatics. International Journal of Food Microbiology, 2006b; 111 (2): 170–4.

● 27. Gouvea F d S, Rosenthal A ,Ferreira EH d R. Plant extract and essential oils added as antimicrobials to cheeses: a review. Microbiology Cienc Rural 2017; 47 (8). https://doi.org/10.1590/0103-8478cr20160908Ciência

● 28. Oliveira TLCd, Ramos ALS, Ramos EM, Piccoli RH, Cristianini M. Natural antimicrobials as additional hurdles to preservation of foods by high pressure processing. Trends in Food Science and Tecnhology 2015; 45 (1): 60–85.

● 29. Govaris A, Botsoglou E, Sergelidis D, Chatzopoulou PS.Antibacterial activity of oregano and thyme essential oils against Listeria monocytogenes and Escherichia coli O157:H7 in Feta cheese packaged under modified atmosphere. LWT - Food Science and Technology, 2011; 44 (4): 1240–4.

● 30. Makkia Dalia I, Mahmoud A H, Bahout A A, Bayoumi M A, Alnakip M E. Molecular Studies on Some Emerging Pathogens in Dairy Products Retailed in Dakahlia Governorate, Egypt.Journal of Advanced Veterinary Research 2022; 12 (4): 392–8.

● 31. Celikel N, Kavas G. Antimicrobial properties of some essential oils against some pathogenic microorganisms. Czech J Food Sci 2008; 26 (3): 174–81.

● 32. El-Khawas K.M. , Hassan H.M. Control of food poisoning bacteria during manufacturing of acid cheese using some organic acids. Assiut Vet Med J 2015; 145: 40–6.

● 33. Chandan RC. Soft and special varieties. In: Cavellaro B, Trugo LC,Finglas PM (eds). Encyclopedia of Food Sciences and Nutrition Vol .2 New York: Academic Press, 2003; 1093–8.

● 34. APHA .American Public Health Association. Compendium of methods for the microbiological examinations of foods. 2nd ed. APHA, Washington, DC, USA. 1992.

● 35. Khalafalla F A, Ali F H, Hassan A R H. Quality improvement and shelf-life extension of refrigerated Nile tilapia (Oreochromis niloticus) fillets using natural herbs. Beni-Suef University J of Basic and Applied Sciences 2015; 4 (1): 33–40.

● 36. Pajohi M R, Tajik H, Farshid AA , Hadian M. Synergistic antibacterial activity of the essential oil of Cuminum cyminum L. seed and nisin in a food model. J of Applied Microbiology 2011; 110 (4): 943–51.

● 37. FDA "Food and Drug Administration" .Bad bug book: Foodborne pathogenic 358 microorganisms and natural toxins handbook, 2nd ed. US Food and Drug 359 Administration, Silver Spring 2012; 87–92.

● 38. Salfinger Y, Tortorello ML Fifth (Ed.) Compendium of Methods for 453 the Microbiological Examination of Foods, 5th Ed., American Public Health 454 Association, Washington, D.C. 2015.

● 39. Feldman D, Hoffman R , Simpson J. The solution for data analysis and presentation graphics. 2nd Ed. Abacus Landcripts, Inc., Barkeley, CA, USA (2003) .

● 40. Virpari PK, Virpari JB, Nayak M N, Thaker HC. Study on isolation, molecular detection of virulence gene and antibiotic sensitivity pattern of Escherichia coli isolated from milk and milk products 2013; 6 (8):541–5.

● 41. Nazzaro F, Fratianni F, De Martino L, Coppola R, De Feo V. Effect of essential oils on pathogenic bacteria. Pharma 2013; 6: 1451–4.

● 42. Boskovic M, Zdravkovic N, Ivanovic J, Janjic J, Djordjevic J, Starcevic M, Baltic M Z. Antimicrobial activity of Thyme (Tymus vulgaris) and oregano (Origanum vulgare) essential oils against some food-borne microorganisms. Procedia Food Science 2015; 5: 18–21.

● 43. Hyldgaard M, Mygind T, Meyer R L. Essential oils in food preservation: Mode of action, synergies, and interactions with food matrix components. Frontiers in Microbiology 2012; 3: 1–24.

● 44. Silva B N, Cadavez V, Teixeira J A , Gonzales-Barron U. Effects of Essential Oils on Escherichia coli Inactivation in Cheese as Described by Meta-Regression Modelling. Foods 2020; 9 (6), 716.

● 45. Olaimat A N, Al-Holy M A, Abu Ghoush M A, Al-Nabulsi A A, Osaili T M, Holley R A. Inhibitory effects of cinnamon and thyme essential oils against Salmonella spp. in hummus (chickpea dip). Journal of Food Processing and Preservation 2019; 43 (5), e13925.

● 46. Koluman A, Dikici A. Antimicrobial resistance of emerging foodborne pathogens: Status quo and global trends. Crit Rev Microbiol 2013; 39 (1), 57–69.

● 47. Anžlovar S, Baričevič D, Ambrožič Avguštin J, Dolenc Koce J. Essential oil of common thyme as a natural antimicrobial food additive. Food Technology and Biotechnology 2014; 52 (2): 263–8.

● 48. EL- Kholy Wedad M, Aamer Reda A , Mailam M A. Effect of Some Essential Oils on the Quality of UF-Soft Cheese During Storage Alex J Fd Sci and Technol 2017; 14(1): 13–27.

● 49. Chacَn MR, Figueras MJ, Castro-Escarpulli G. Soler L, Guarro J .Distribution of virulence genes in clinical and environmental isolates of Aeromonas spp. Antonie Van Leeuwenhoek 2003; 84 (4): 269–8.

● 50. Hoel S, Vadstein O, Jakobsen AN. Species distribution and prevalence of putative virulence factors in Mesophilic Aeromonas spp. isolated from fresh retail sushi. Front. Microbiol. 2017; 8, 931.

● 51. Yang Y, Miao P, Li H, Tan S, Yu H, Yu H. Antibiotic susceptibility and molecular characterization of Aeromonas hydrophila from grass carp. J Food Saf 2018; 38 (1), e12393.

● 52. Kirrella Ghada A K, Moustafa Nader Y , Kishk Dina M, Abdallah Reda. Impact of using some essential oils on sensory acceptability and Aeromonas hydrophila contamination of Nile tilapia fish fillet during refrigeration storage KVMJ 2021; 19 (2):9–13.

● 53. Wang J, Stanford K, McAllister TA, Johnson RP, Chen J, Hou H, Zhang G, Niu YD. Biofilm formation, virulence gene profiles, and antimicrobial resis¬tance of nine serogroups of non-O157 Shiga toxin-pro¬ducing Escherichia coli. Foodborne Pathogens and Diseases 2016; 13 (6): 316–24.

● 54. Logue CM, Doetkott C, Mangiamele P. Genotypic and phenotypic traits that distinguish neonatal meningitis-associated Escherichia coli from fecal E. coli isolates of healthy human hosts. Appl Environ. Microbiol 2012; 78 (16): 5824-–30.

● 55. Ibrahim EMA, Elbarbary HA, Shawky NA, El-Sebay IA. Survival of Shiga-toxin Producing Escherichia coli (STEC) during storage of Karish cheese. AJVS 2020; 64 (1): 47–53.

● 56. Hussien Heba, Elbehiry A, Saad Marwa, Hadad Ghada, Moussa I, Dawoud T, Mubarak A, Marzouk Eman. Molecular characterization of Escherichia coli isolated from cheese and biocontrol of Shiga toxigenic E. coli with essential oils. Ital J of Food Safety 2019; 8: 8291.

● 57. Salem Amani M, Abo El-Roos Nahla A, Abd EL-Fatah Mona M. Improvement quality of minced meat using some essential oils. Benha Veterinary Medical J 2019; 37(1) :35–65.

● 58. Burt SA, Reinders RD. Antibacterial activity of selected plant essential oils against Escherichia coli O157:H7. Lett Appl Microbiol. 2003; 36 (3): 162–7.

● 59. Mahboubi M, Heidarytabar R, Mahdizadeh E, Hosseini H. Antimicrobial activity and chemical composition of Thymus species and Zataria multiflora essential oils. Agriculture and Natural Resources 2017; 51 (5): 395–401.

● 60. Bourgou S, Pichette A, Marzouk B , Legault J. Antioxidant, anti-inflammatory, anticancer and antibacterial activities of extracts from Nigella Sativa (Black Cumin) plant parts. J Food Bio-chem. 2012; 36:539–6.

● 61. Al-Nabulsi A, Osaili T, Olaimat A, Almasri Weam Al-Holy M, Jaradat Z, Ayyash M, Awaisheh S, Holley R. Inhibitory effect of thyme and cinnamon essential oils against E. coli O157:H7 in Tahini. Food Sci and Technol Campinas 2020; 40 (4): 885–93.

● 62. Mani-López E, García H S, López-Malo A. Organic acids as antimicrobials to control Salmonella in meat and poultry products. Food Research International 2012; 45 (2): 713–21.

● 63. Perricone M, Arace E, Corbo M R, Sinigaglia M, Bevilacqua A. Bioactivity of essential oils: A review on their interaction with food components. Frontiers in Microbiology 2015; 6: 1–16.

● 64. Oussalah M, Caillet S, Saucier L, Lacroix M. Inhibitory effects of selected plant essential oils on the growth of four pathogenic bacteria: E. coli O157:H7, Salmonella typhimurium, Staphylococcus aureus and Listeria monocytogenes. Food Control 2007; 18 (5): 414–20.

● 65. Hossain M A, AL-Raqmi K A S, AL-Mijizy Z H, Weli A M, Al-Riyami Q. Study of total phenol, flavonoids contents and phytochemical screening of various leaves crude extracts of locally grown Thymus vulgaris. Asian Pacific J of Tropical Biomedicine, 2013; 3: 705–10.

● 66. Selim S. Antimicrobial Activity Of Essential Oils Against Vancomycin-Resistant Enterococci (Vre) And Escherichia Coli O157:H7 In Feta Soft Cheese And Minced Beef Meat. Brazilian J of Microbiol 2011; 42: 187–96.

● 67. Alsawaf S D, Alnaemi H S. Effect of Nigella sativa (seed and oil) on the bacteriological quality of soft white cheese. Iraqi J of Veterinary Sciences 2011; 251: 21–7.

● 68. Al-Salman H K. Antibacterial and antifungal activity of volatile oils of some medicinal plants. Iraqi J. Desert Studies 2008; 1: 91–103.




How to Cite

Makkia, D., Bahout, A., Bayoumi, M. A., Alnakip, M. E., & Moustafa, A. H. (2023). EFFECT OF ESSENTIAL OILS ON MULTIDRUG RESISTANT GRAM-NEGATIVE BACTERIA. Slovenian Veterinary Research, 60(25-Suppl), 149–57. https://doi.org/10.26873/SVR-1574-2022



Veterinary Medicine and The One Health Concept