Applied Microbiology and Biotechnology

, Volume 102, Issue 23, pp 10209–10218 | Cite as

Effect of thyme essential oil against Bacillus cereus planktonic growth and biofilm formation

  • Jiamu Kang
  • Liu LiuEmail author
  • Xiaoxia Wu
  • Yuyang Sun
  • Zifei Liu
Applied microbial and cell physiology


The objective of this study was to determine the effect of thyme essential oil (TEO) on the planktonic growth and biofilm formation of Bacillus cereus (B. cereus). GC-MS analysis of TEO allowed the detection of 13 compounds, and the major constituents were p-cymene (29.7%), thymol (23.73%), γ-terpinene (16.21%), and 1,8-cineole (9.74%). TEO exhibited a minimum inhibitory concentration (MIC) value against planktonic B. cereus of 0.25 mg/mL. The potent effect of TEO to inhibit the growth of planktonic B. cereus was due to cell membrane damage, as evidenced by reduced cell viability, protein changes, decreased intracellular ATP concentration, increased extracellular ATP concentration and cell membrane depolarization, and cellular morphological changes. In addition, TEO exerted a significant inhibitory effect on B. cereus biofilm formation, as confirmed by environmental scanning electron microscopic images. These findings suggested that TEO has the potential to be developed as a natural food additive to control foodborne contamination associated with B. cereus and its biofilm.


Thyme essential oil Bacillus cereus Cell membrane Biofilm formation 



This research was financially supported by the National Natural Science Foundations of China (Grant No. 31301472).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.


  1. Abdollahzadeh E, Rezaei M, Hosseini H (2014) Antibacterial activity of plant essential oils and extracts: the role of thyme essential oil, nisin, and their combination to control Listeria monocytogenes inoculated in minced fish meat. Food Control 35(1):177–183. CrossRefGoogle Scholar
  2. Adukwu EC, Bowles M, Edwards-Jones V, Bone H (2016) Antimicrobial activity, cytotoxicity and chemical analysis of lemongrass essential oil (Cymbopogon flexuosus) and pure citral. Appl Microbiol Biotechnol 100(22):9619–9627. CrossRefPubMedPubMedCentralGoogle Scholar
  3. Anderson Borge GI, Skeie M, Sørhaug T, Langsrud T, Granum PE (2001) Growth and toxin profiles of Bacillus cereus isolated from different food sources. Int J Food Microbiol 69(3):237–246CrossRefGoogle Scholar
  4. Auger S, Krin E, Aymerich S, Gohar M (2006) Autoinducer 2 affects biofilm formation by Bacillus cereus. Appl Environ Microbiol 72(1):937–941. CrossRefPubMedPubMedCentralGoogle Scholar
  5. Bagamboula CF, Uyttendaele M, Debevere J (2004) Inhibitory effect of thyme and basil essential oils, carvacrol, thymol, estragol, linalool and p-cymene towards Shigella sonnei and S. flexneri. Food Microbiol 21(1):33–42. CrossRefGoogle Scholar
  6. Bajpai VK, Sharma A, Baek K-H (2013) Antibacterial mode of action of Cudrania tricuspidata fruit essential oil, affecting membrane permeability and surface characteristics of food-borne pathogens. Food Control 32(2):582–590. CrossRefGoogle Scholar
  7. Boskovic M, Djordjevic J, Ivanovic J, Janjic J, Zdravkovic N, Glisic M, Glamoclija N, Baltic B, Djordjevic V, Baltic M (2017) Inhibition of Salmonella by thyme essential oil and its effect on microbiological and sensory properties of minced pork meat packaged under vacuum and modified atmosphere. Int J Food Microbiol 258:58–67. CrossRefPubMedGoogle Scholar
  8. Burt S (2004) Essential oils: their antibacterial properties and potential applications in foods – a review. Int J Food Microbiol 94(3):223–253. CrossRefPubMedGoogle Scholar
  9. Calo JR, Crandall PG, O'Bryan CA, Ricke SC (2015) Essential oils as antimicrobials in food systems – a review. Food Control 54:111–119. CrossRefGoogle Scholar
  10. Cui HY, Zhou H, Lin L, Zhao CT, Zhang XJ, Xiao ZH, Li CZ (2016) Antibacterial activity and mechanism of cinnamon essential oil and its application in milk. J Anim Plant Sci 26(2):533–541Google Scholar
  11. Debonne E, Van Bockstaele F, De Leyn I, Devlieghere F, Eeckhout M (2018) Validation of in-vitro antifungal activity of thyme essential oil on Aspergillus niger and Penicillium paneum through application in par-baked wheat and sourdough bread. LWT Food Sci Technol 87:368–378. CrossRefGoogle Scholar
  12. Edris AE (2007) Pharmaceutical and therapeutic potentials of essential oils and their individual volatile constituents: a review. Phytother Res 21(4):308–323. CrossRefPubMedGoogle Scholar
  13. EFSA (2005) Opinion of the scientific panel on biological hazards (BIOHAZ) on Bacillus cereus and other Bacillus spp in foodstuffs. EFSA J 175:1–48Google Scholar
  14. Ehling-Schulz M, Fricker M, Scherer S (2004) Bacillus cereus, the causative agent of an emetic type of food-borne illness. Mol Nutr Food Res 48(7):479–487. CrossRefPubMedGoogle Scholar
  15. Ghabraie M, Vu KD, Tata L, Salmieri S, Lacroix M (2016) Antimicrobial effect of essential oils in combinations against five bacteria and their effect on sensorial quality of ground meat. LWT Food Sci Technol 66:332–339. CrossRefGoogle Scholar
  16. Hayrapetyan H, Muller L, Tempelaars M, Abee T, Nierop Groot M (2015) Comparative analysis of biofilm formation by Bacillus cereus reference strains and undomesticated food isolates and the effect of free iron. Int J Food Microbiol 200:72–79. CrossRefPubMedGoogle Scholar
  17. Houry A, Briandet R, Aymerich S, Gohar M (2010) Involvement of motility and flagella in Bacillus cereus biofilm formation. Microbiology 156(Pt 4):1009–1018. CrossRefPubMedGoogle Scholar
  18. Jouki M, Yazdi FT, Mortazavi SA, Koocheki A, Khazaei N (2014) Effect of quince seed mucilage edible films incorporated with oregano or thyme essential oil on shelf life extension of refrigerated rainbow trout fillets. Int J Food Microbiol 174:88–97. CrossRefPubMedGoogle Scholar
  19. Kang J, Li Q, Liu L, Jin W, Wang J, Sun Y (2018) The specific effect of gallic acid on Escherichia coli biofilm formation by regulating pgaABCD genes expression. Appl Microbiol Biotechnol 102(4):1837–1846. CrossRefPubMedGoogle Scholar
  20. Karunakaran E, Biggs CA (2011) Mechanisms of Bacillus cereus biofilm formation: an investigation of the physicochemical characteristics of cell surfaces and extracellular proteins. Appl Microbiol Biotechnol 89(4):1161–1175. CrossRefPubMedGoogle Scholar
  21. Kumari S, Sarkar PK (2016) Bacillus cereus hazard and control in industrial dairy processing environment. Food Control 69:20–29. CrossRefGoogle Scholar
  22. Li G, Xu Y, Wang X, Zhang B, Shi C, Zhang W, Xia X (2014) Tannin-rich fraction from pomegranate rind damages membrane of Listeria monocytogenes. Foodborne Pathog Dis 11(4):313–319. CrossRefPubMedGoogle Scholar
  23. Li WR, Li HL, Shi QS, Sun TL, Xie XB, Song B, Huang XM (2016) The dynamics and mechanism of the antimicrobial activity of tea tree oil against bacteria and fungi. Appl Microbiol Biotechnol 100(20):8865–8875. CrossRefPubMedGoogle Scholar
  24. Liu F, Wang F, Du L, Zhao T, Doyle MP, Wang D, Zhang X, Sun Z, Xu W (2018) Antibacterial and antibiofilm activity of phenyllactic acid against Enterobacter cloacae. Food Control 84:442–448. CrossRefGoogle Scholar
  25. Lv F, Liang H, Yuan Q, Li C (2011) In vitro antimicrobial effects and mechanism of action of selected plant essential oil combinations against four food-related microorganisms. Food Res Int 44(9):3057–3064. CrossRefGoogle Scholar
  26. Miguel MG (2010) Antioxidant and anti-inflammatory activities of essential oils: a short review. Molecules 15(12):9252–9287. CrossRefPubMedGoogle Scholar
  27. Parke DV, Lewis DF (1992) Safety aspects of food preservatives. Food Addit Contam 9(5):561–577. CrossRefPubMedGoogle Scholar
  28. Randazzo CL, Scifo GO, Tomaselli F, Caggia C (2009) Polyphasic characterization of bacterial community in fresh cut salads. Int J Food Microbiol 128(3):484–490. CrossRefPubMedGoogle Scholar
  29. Sadekuzzaman M, Mizan MFR, Kim H-S, Yang S, Ha S-D (2018) Activity of thyme and tea tree essential oils against selected foodborne pathogens in biofilms on abiotic surfaces. LWT Food Sci Technol 89:134–139. CrossRefGoogle Scholar
  30. Samapundo S, Heyndrickx M, Xhaferi R, Devlieghere F (2011) Incidence, diversity and toxin gene characteristics of Bacillus cereus group strains isolated from food products marketed in Belgium. Int J Food Microbiol 150(1):34–41. CrossRefPubMedGoogle Scholar
  31. Segvic Klaric M, Kosalec I, Mastelic J, Pieckova E, Pepeljnak S (2007) Antifungal activity of thyme (Thymus vulgaris L.) essential oil and thymol against moulds from damp dwellings. Lett Appl Microbiol 44(1):36–42. CrossRefPubMedGoogle Scholar
  32. Shi C, Song K, Zhang X, Sun Y, Sui Y, Chen Y, Jia Z, Sun H, Sun Z, Xia X (2016) Antimicrobial activity and possible mechanism of action of Citral against Cronobacter sakazakii. PLoS One 11(7):e0159006. CrossRefPubMedPubMedCentralGoogle Scholar
  33. Singh N, Singh RK, Bhunia AK, Stroshine RL (2002) Efficacy of chlorine dioxide, ozone, and thyme essential oil or a sequential washing in killing Escherichia coli O157:H7 on lettuce and baby carrots. LWT Food Sci Technol 35(8):720–729. CrossRefGoogle Scholar
  34. Solomakos N, Govaris A, Koidis P, Botsoglou N (2008) The antimicrobial effect of thyme essential oil, nisin and their combination against Escherichia coli O157:H7 in minced beef during refrigerated storage. Meat Sci 80(2):159–166. CrossRefPubMedGoogle Scholar
  35. Stiefel P, Schmidt-Emrich S, Maniura-Weber K, Ren Q (2015) Critical aspects of using bacterial cell viability assays with the fluorophores SYTO9 and propidium iodide. BMC Microbiol 15:36. CrossRefPubMedPubMedCentralGoogle Scholar
  36. Tohidi B, Rahimmalek M, Arzani A (2017) Essential oil composition, total phenolic, flavonoid contents, and antioxidant activity of Thymus species collected from different regions of Iran. Food Chem 220:153–161. CrossRefPubMedGoogle Scholar
  37. Valero M, Salmerón MC (2003) Antibacterial activity of 11 essential oils against Bacillus cereus in tyndallized carrot broth. Int J Food Microbiol 85(1–2):73–81. CrossRefPubMedGoogle Scholar
  38. Valero M, Hernandez-Herrero LA, Giner MJ (2007) Survival, isolation and characterization of a psychrotrophic Bacillus cereus strain from a mayonnaise-based ready-to-eat vegetable salad. Food Microbiol 24(7–8):671–677. CrossRefPubMedGoogle Scholar
  39. Van Haute S, Raes K, Van der Meeren P, Sampers I (2016) The effect of cinnamon, oregano and thyme essential oils in marinade on the microbial shelf life of fish and meat products. Food Control 68:30–39. CrossRefGoogle Scholar
  40. Wang C, Chang T, Yang H, Cui M (2015) Antibacterial mechanism of lactic acid on physiological and morphological properties of Salmonella Enteritidis, Escherichia coli and Listeria monocytogenes. Food Control 47:231–236. CrossRefGoogle Scholar
  41. Wijman JG, de Leeuw PP, Moezelaar R, Zwietering MH, Abee T (2007) Air-liquid interface biofilms of Bacillus cereus: formation, sporulation, and dispersion. Appl Environ Microbiol 73(5):1481–1488. CrossRefPubMedPubMedCentralGoogle Scholar
  42. Xu C, Li J, Yang L, Shi F, Yang L, Ye M (2017) Antibacterial activity and a membrane damage mechanism of Lachnum YM30 melanin against Vibrio parahaemolyticus and Staphylococcus aureus. Food Control 73:1445–1451. CrossRefGoogle Scholar
  43. Zhang Y, Liu X, Wang Y, Jiang P, Quek S (2016) Antibacterial activity and mechanism of cinnamon essential oil against Escherichia coli and Staphylococcus aureus. Food Control 59:282–289. CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Jiamu Kang
    • 1
  • Liu Liu
    • 1
    Email author
  • Xiaoxia Wu
    • 1
  • Yuyang Sun
    • 1
  • Zifei Liu
    • 1
  1. 1.College of Food Engineering and Nutrition ScienceShaanxi Normal UniversityXi’anChina

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