Abstract
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.
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References
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. https://doi.org/10.1016/j.foodcont.2013.07.004
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. https://doi.org/10.1007/s00253-016-7807-y
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):237–246
Auger S, Krin E, Aymerich S, Gohar M (2006) Autoinducer 2 affects biofilm formation by Bacillus cereus. Appl Environ Microbiol 72(1):937–941. https://doi.org/10.1128/AEM.72.1.937-941.2006
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. https://doi.org/10.1016/s0740-0020(03)00046-7
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. https://doi.org/10.1016/j.foodcont.2013.01.032
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. https://doi.org/10.1016/j.ijfoodmicro.2017.07.011
Burt S (2004) Essential oils: their antibacterial properties and potential applications in foods – a review. Int J Food Microbiol 94(3):223–253. https://doi.org/10.1016/j.ijfoodmicro.2004.03.022
Calo JR, Crandall PG, O'Bryan CA, Ricke SC (2015) Essential oils as antimicrobials in food systems – a review. Food Control 54:111–119. https://doi.org/10.1016/j.foodcont.2014.12.040
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):533–541
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. https://doi.org/10.1016/j.lwt.2017.09.007
Edris AE (2007) Pharmaceutical and therapeutic potentials of essential oils and their individual volatile constituents: a review. Phytother Res 21(4):308–323. https://doi.org/10.1002/ptr.2072
EFSA (2005) Opinion of the scientific panel on biological hazards (BIOHAZ) on Bacillus cereus and other Bacillus spp in foodstuffs. EFSA J 175:1–48
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. https://doi.org/10.1002/mnfr.200400055
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. https://doi.org/10.1016/j.lwt.2015.10.055
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. https://doi.org/10.1016/j.ijfoodmicro.2015.02.005
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. https://doi.org/10.1099/mic.0.034827-0
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. https://doi.org/10.1016/j.ijfoodmicro.2014.01.001
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. https://doi.org/10.1007/s00253-017-8709-3
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. https://doi.org/10.1007/s00253-010-2919-2
Kumari S, Sarkar PK (2016) Bacillus cereus hazard and control in industrial dairy processing environment. Food Control 69:20–29. https://doi.org/10.1016/j.foodcont.2016.04.012
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. https://doi.org/10.1089/fpd.2013.1675
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. https://doi.org/10.1007/s00253-016-7692-4
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. https://doi.org/10.1016/j.foodcont.2017.09.004
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. https://doi.org/10.1016/j.foodres.2011.07.030
Miguel MG (2010) Antioxidant and anti-inflammatory activities of essential oils: a short review. Molecules 15(12):9252–9287. https://doi.org/10.3390/molecules15129252
Parke DV, Lewis DF (1992) Safety aspects of food preservatives. Food Addit Contam 9(5):561–577. https://doi.org/10.1080/02652039209374110
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. https://doi.org/10.1016/j.ijfoodmicro.2008.10.019
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. https://doi.org/10.1016/j.lwt.2017.10.042
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. https://doi.org/10.1016/j.ijfoodmicro.2011.07.013
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. https://doi.org/10.1111/j.1472-765X.2006.02032.x
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. https://doi.org/10.1371/journal.pone.0159006
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. https://doi.org/10.1006/fstl.2002.0933
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. https://doi.org/10.1016/j.meatsci.2007.11.014
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. https://doi.org/10.1186/s12866-015-0376-x
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. https://doi.org/10.1016/j.foodchem.2016.09.203
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. https://doi.org/10.1016/s0168-1605(02)00484-1
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. https://doi.org/10.1016/j.fm.2007.04.005
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. https://doi.org/10.1016/j.foodcont.2016.03.025
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. https://doi.org/10.1016/j.foodcont.2014.06.034
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. https://doi.org/10.1128/AEM.01781-06
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. https://doi.org/10.1016/j.foodcont.2016.10.048
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. https://doi.org/10.1016/j.foodcont.2015.05.032
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This research was financially supported by the National Natural Science Foundations of China (Grant No. 31301472).
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Kang, J., Liu, L., Wu, X. et al. Effect of thyme essential oil against Bacillus cereus planktonic growth and biofilm formation. Appl Microbiol Biotechnol 102, 10209–10218 (2018). https://doi.org/10.1007/s00253-018-9401-y
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DOI: https://doi.org/10.1007/s00253-018-9401-y