Abstract
Introduction: This study focused on the formulation processing and antimicrobial activities of Origanum and Thyme essential oils (OEO) (TEO) and their nanoemulsions (OEON) (TEON) on gram-positive and gram-negative bacteria and yeast. Optimal nanoemulsion formulation and stability were achieved through balancing between requisite hydrophilic-lipophilic balance (rHLB) and surfactant concentration (Smix) of nanoemulsions.
Methods: The smallest droplet sizes of OEON: (z -185.1 nm ± 0.85) and TEON (z -130.1 nm ± 0.60), were achieved by using 10 g/100 mL of OEO and TEO with 15 g/100 mL Smix at ultrasonication for 2 min with no phase separation. The size of nanoemulsion droplet and PDI was found to be influenced by HLB value, treatment time, and Smix concentration. Antimicrobial analyses, including a zone of inhibition, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and time-kill kinetics assay, were performed against Escherichia coli, Staphylococcus aureus, and Saccharomyces cerevisiae.
Results: The antimicrobial findings demonstrated that OEO exhibited higher antimicrobial activity compared to TEO (P<0.05). Meanwhile, the OEON and TEON substantially reduced the MIC and MBC values, compared to OEO and TEO against all tested microorganisms (P<0.05). Throughout the time-kill assay, E. coli was reduced by 5 log CFU/ml within 120 minutes, while S. cerevisiae and S. aureus were eradicated after 60 and 120 minutes of incubation with OEON, respectively.
Conclusion: In turn, TEON reduced E. coli by 5 log CFU/ml and S. cerevisiae by 4 log CFU/ml within 120 minutes, while S. aureus was inhibited within the same time of incubation with TEON. The nanoemulsion formulations of OEO and TEO considerably enhanced the antimicrobial properties, which provides a promising alternative plant-derived antimicrobial for pharmaceuticals and food applications.
Graphical Abstract
[1]
Zawani CJ, Nor-Khaizura MAR, Mahyudin NA, Ismail-Fitry MR, Nirmal NP. Microbiological and sensorial quality of beef meat (Longissimus dorsi) marinated with cinnamon extract and stored at various temperatures. Foods 2022; 11(24): 3971.
[http://dx.doi.org/10.3390/foods11243971] [PMID: 36553712]
[http://dx.doi.org/10.3390/foods11243971] [PMID: 36553712]
[2]
Nirmal NP, Khanashyam AC, Mundanat AS, et al. Valorization of fruit waste for bioactive compounds and their applications in the food industry. Foods 2023; 12(3): 556.
[http://dx.doi.org/10.3390/foods12030556] [PMID: 36766085]
[http://dx.doi.org/10.3390/foods12030556] [PMID: 36766085]
[3]
Nguyen CNM, Nirmal NP, Sultanbawa Y, Ziora ZM. Antioxidant and antibacterial activity of four tannins isolated from different sources and their effect on the shelf-life extension of vacuum packed minced meat. Foods 2023; 12(2): 354.
[http://dx.doi.org/10.3390/foods12020354] [PMID: 36673446]
[http://dx.doi.org/10.3390/foods12020354] [PMID: 36673446]
[4]
Moghimi R, Aliahmadi A, McClements DJ, Rafati H. Investigations of the effectiveness of nanoemulsions from sage oil as antibacterial agents on some food borne pathogens. Lebensm Wiss Technol 2016; 71: 69-76.
[http://dx.doi.org/10.1016/j.lwt.2016.03.018]
[http://dx.doi.org/10.1016/j.lwt.2016.03.018]
[5]
Nirmal NP, Mereddy R, Li L, Sultanbawa Y. Formulation, characterisation and antibacterial activity of lemon myrtle and anise myrtle essential oil in water nanoemulsion. Food Chem 2018; 254: 1-7.
[http://dx.doi.org/10.1016/j.foodchem.2018.01.173] [PMID: 29548427]
[http://dx.doi.org/10.1016/j.foodchem.2018.01.173] [PMID: 29548427]
[6]
Hassoun A, Emir Çoban Ö. Essential oils for antimicrobial and antioxidant applications in fish and other seafood products. Trends Food Sci Technol 2017; 68: 26-36.
[http://dx.doi.org/10.1016/j.tifs.2017.07.016]
[http://dx.doi.org/10.1016/j.tifs.2017.07.016]
[7]
Dávila-Rodríguez M, López-Malo A, Palou E, Ramírez-Corona N, Jiménez-Munguía MT. Antimicrobial activity of nanoemulsions of cinnamon, rosemary, and oregano essential oils on fresh celery. Lebensm Wiss Technol 2019; 112: 108247.
[http://dx.doi.org/10.1016/j.lwt.2019.06.014]
[http://dx.doi.org/10.1016/j.lwt.2019.06.014]
[8]
Nirmal NP, Chunhavacharatorn P, Chandra K. A, Li L, Al-Asmari F. Cinnamon bark oil in water nanoemulsion formulation, characterization, and antimicrobial activities. Lebensm Wiss Technol 2023; 179: 114671.
[http://dx.doi.org/10.1016/j.lwt.2023.114671]
[http://dx.doi.org/10.1016/j.lwt.2023.114671]
[9]
Jawaid T, Alaseem AM, Khan MM, et al. Preparation and evaluation of nanoemulsion of citronella essential oil with improved antimicrobial and anti-cancer properties. Antibiotics 2023; 12(3): 478.
[http://dx.doi.org/10.3390/antibiotics12030478] [PMID: 36978345]
[http://dx.doi.org/10.3390/antibiotics12030478] [PMID: 36978345]
[10]
Sun X, Wang J, Zhang H, et al. Development of functional gelatin-based composite films incorporating oil-in-water lavender essential oil nano-emulsions: Effects on physicochemical properties and cherry tomatoes preservation. Lebensm Wiss Technol 2021; 142: 110987.
[http://dx.doi.org/10.1016/j.lwt.2021.110987]
[http://dx.doi.org/10.1016/j.lwt.2021.110987]
[11]
Jiang H, Zhong S, Schwarz P, Chen B, Rao J. Antifungal activity, mycotoxin inhibitory efficacy, and mode of action of hop essential oil nanoemulsion against Fusarium graminearum. Food Chem 2023; 400: 134016.
[http://dx.doi.org/10.1016/j.foodchem.2022.134016] [PMID: 36084588]
[http://dx.doi.org/10.1016/j.foodchem.2022.134016] [PMID: 36084588]
[12]
Watcharin W, Gupta S, Saning A, Laodheerasiri S, Chuenchom L. Free radical scavenging effects of grapefruit essential oil nanoemulsion stabilized with carrageenan and its cytotoxicity assay on HeLa cell line. Adv Nat Sci: Nanosci Nanotechnol 2023; 14(2): 025014.
[http://dx.doi.org/10.1088/2043-6262/acd6e5]
[http://dx.doi.org/10.1088/2043-6262/acd6e5]
[13]
Mushtaq A, Mohd Wani S, Malik AR, et al. Recent insights into Nanoemulsions: Their preparation, properties and applications. Food Chem X 2023; 18: 100684.
[http://dx.doi.org/10.1016/j.fochx.2023.100684] [PMID: 37131847]
[http://dx.doi.org/10.1016/j.fochx.2023.100684] [PMID: 37131847]
[14]
Rathod NB, Meral R, Siddiqui SA, Nirmal N, Ozogul F. Nanoemulsion-based approach to preserve muscle food: A review with current knowledge. Crit Rev Food Sci Nutr 2023; 1-22.
[http://dx.doi.org/10.1080/10408398.2023.2175347] [PMID: 36789616]
[http://dx.doi.org/10.1080/10408398.2023.2175347] [PMID: 36789616]
[15]
Ansarifar E, Moradinezhad F. Encapsulation of thyme essential oil using electrospun zein fiber for strawberry preservation. Chem Biol Technol Agric 2022; 9(1): 2.
[http://dx.doi.org/10.1186/s40538-021-00267-y]
[http://dx.doi.org/10.1186/s40538-021-00267-y]
[16]
Veenstra JP, Johnson JJ. Oregano (Origanum vulgare) extract for food preservation and improvement in gastrointestinal health. Int J Nutr 2019; 3(4): 43-52.
[http://dx.doi.org/10.14302/issn.2379-7835.ijn-19-2703] [PMID: 31080888]
[http://dx.doi.org/10.14302/issn.2379-7835.ijn-19-2703] [PMID: 31080888]
[17]
Lee S, Kim H, Beuchat LR, Kim Y, Ryu JH. Synergistic antimicrobial activity of oregano and thyme thymol essential oils against Leuconostoc citreum in a laboratory medium and tomato juice. Food Microbiol 2020; 90: 103489.
[http://dx.doi.org/10.1016/j.fm.2020.103489] [PMID: 32336377]
[http://dx.doi.org/10.1016/j.fm.2020.103489] [PMID: 32336377]
[18]
Rathod NB, Kulawik P, Ozogul F, Regenstein JM, Ozogul Y. Biological activity of plant-based carvacrol and thymol and their impact on human health and food quality. Trends Food Sci Technol 2021; 116: 733-48.
[http://dx.doi.org/10.1016/j.tifs.2021.08.023]
[http://dx.doi.org/10.1016/j.tifs.2021.08.023]
[19]
Burt S. Essential oils: Their antibacterial properties and potential applications in foods—a review. Int J Food Microbiol 2004; 94(3): 223-53.
[http://dx.doi.org/10.1016/j.ijfoodmicro.2004.03.022] [PMID: 15246235]
[http://dx.doi.org/10.1016/j.ijfoodmicro.2004.03.022] [PMID: 15246235]
[20]
Sandhya M, Ramasamy D, Sudhakar K, Kadirgama K, Harun WSW. Ultrasonication an intensifying tool for preparation of stable nanofluids and study the time influence on distinct properties of graphene nanofluids – A systematic overview. Ultrason Sonochem 2021; 73: 105479.
[http://dx.doi.org/10.1016/j.ultsonch.2021.105479] [PMID: 33578278]
[http://dx.doi.org/10.1016/j.ultsonch.2021.105479] [PMID: 33578278]
[21]
Badr MM, Badawy MEI, Taktak NEM. Preparation, characterization, and antimicrobial activity of cinnamon essential oil and cinnamaldehyde nanoemulsions. J Essent Oil Res 2022; 34(6): 544-58.
[http://dx.doi.org/10.1080/10412905.2022.2107100]
[http://dx.doi.org/10.1080/10412905.2022.2107100]
[22]
NCCLS. Performance standards for antimicrobial susceptibility testing. Ninth informational supplement NCCLS document M100-S9 National committee for clinical laboratory standards. 120-7.
[23]
Rodrigues RF, Costa IC, Almeida FB, et al. Development and characterization of evening primrose (Oenothera biennis) oil nanoemulsions. Rev Bras Farmacogn 2015; 25(4): 422-5.
[http://dx.doi.org/10.1016/j.bjp.2015.07.014]
[http://dx.doi.org/10.1016/j.bjp.2015.07.014]
[24]
Jasmina H, Džana O, Alisa E, Edina V, Ognjenka R, Eds. Preparation of nanoemulsions by high-energy and lowenergy emulsification methods. CMBEBIH 2017: Proceedings of the International Conference on Medical and Biological Engineering.
[http://dx.doi.org/10.1007/978-981-10-4166-2_48]
[http://dx.doi.org/10.1007/978-981-10-4166-2_48]
[25]
Pongsumpun P, Iwamoto S, Siripatrawan U. Response surface methodology for optimization of cinnamon essential oil nanoemulsion with improved stability and antifungal activity. Ultrason Sonochem 2020; 60: 104604.
[http://dx.doi.org/10.1016/j.ultsonch.2019.05.021] [PMID: 31539730]
[http://dx.doi.org/10.1016/j.ultsonch.2019.05.021] [PMID: 31539730]
[26]
Donsì F, Ferrari G. Essential oil nanoemulsions as antimicrobial agents in food. J Biotechnol 2016; 233: 106-20.
[http://dx.doi.org/10.1016/j.jbiotec.2016.07.005] [PMID: 27416793]
[http://dx.doi.org/10.1016/j.jbiotec.2016.07.005] [PMID: 27416793]
[27]
Shamsara O, Muhidinov ZK, Jafari SM, et al. Effect of ultrasonication, pH and heating on stability of apricot gum–lactoglobuline two layer nanoemulsions. Int J Biol Macromol 2015; 81: 1019-25.
[http://dx.doi.org/10.1016/j.ijbiomac.2015.09.056] [PMID: 26432369]
[http://dx.doi.org/10.1016/j.ijbiomac.2015.09.056] [PMID: 26432369]
[28]
Rebolleda S, Sanz MT, Benito JM, Beltrán S, Escudero I, González San-José ML. Formulation and characterisation of wheat bran oil-in-water nanoemulsions. Food Chem 2015; 167: 16-23.
[http://dx.doi.org/10.1016/j.foodchem.2014.06.097] [PMID: 25148953]
[http://dx.doi.org/10.1016/j.foodchem.2014.06.097] [PMID: 25148953]
[29]
Salvia-Trujillo L, Rojas-Graü A, Soliva-Fortuny R, Martín-Belloso O. Physicochemical characterization of lemongrass essential oil–alginate nanoemulsions: effect of ultrasound processing parameters. Food Bioprocess Technol 2013; 6(9): 2439-46.
[http://dx.doi.org/10.1007/s11947-012-0881-y]
[http://dx.doi.org/10.1007/s11947-012-0881-y]
[30]
Hashtjin AM, Abbasi S. Nano-emulsification of orange peel essential oil using sonication and native gums. Food Hydrocoll 2015; 44: 40-8.
[http://dx.doi.org/10.1016/j.foodhyd.2014.08.017]
[http://dx.doi.org/10.1016/j.foodhyd.2014.08.017]
[31]
Gaikwad SG, Pandit AB. Ultrasound emulsification: Effect of ultrasonic and physicochemical properties on dispersed phase volume and droplet size. Ultrason Sonochem 2008; 15(4): 554-63.
[http://dx.doi.org/10.1016/j.ultsonch.2007.06.011] [PMID: 17698396]
[http://dx.doi.org/10.1016/j.ultsonch.2007.06.011] [PMID: 17698396]
[32]
Trombetta D, Castelli F, Sarpietro MG, et al. Mechanisms of antibacterial action of three monoterpenes. Antimicrob Agents Chemother 2005; 49(6): 2474-8.
[http://dx.doi.org/10.1128/AAC.49.6.2474-2478.2005] [PMID: 15917549]
[http://dx.doi.org/10.1128/AAC.49.6.2474-2478.2005] [PMID: 15917549]
[33]
Nazzaro F, Fratianni F, De Martino L, Coppola R, De Feo V. Effect of essential oils on pathogenic bacteria. Pharmaceuticals 2013; 6(12): 1451-74.
[http://dx.doi.org/10.3390/ph6121451] [PMID: 24287491]
[http://dx.doi.org/10.3390/ph6121451] [PMID: 24287491]
[34]
Liao W, Badri W, Dumas E, et al. Nanoencapsulation of essential oils as natural food antimicrobial agents: An overview. Appl Sci 2021; 11(13): 5778.
[http://dx.doi.org/10.3390/app11135778]
[http://dx.doi.org/10.3390/app11135778]
[35]
Maryam I, Huzaifa U, Hindatu H, Zubaida S. Nanoencapsulation of essential oils with enhanced antimicrobial activity: A new way of combating antimicrobial Resistance. J Pharmacogn Phytochem 2015; 4(3): 165-70.
[36]
Salvia-Trujillo L, Rojas-Graü A, Soliva-Fortuny R, Martín-Belloso O. Physicochemical characterization and antimicrobial activity of food-grade emulsions and nanoemulsions incorporating essential oils. Food Hydrocoll 2015; 43: 547-56.
[http://dx.doi.org/10.1016/j.foodhyd.2014.07.012]
[http://dx.doi.org/10.1016/j.foodhyd.2014.07.012]
[37]
Li W, Chen H, He Z, Han C, Liu S, Li Y. Influence of surfactant and oil composition on the stability and antibacterial activity of eugenol nanoemulsions. Lebensm Wiss Technol 2015; 62(1): 39-47.
[http://dx.doi.org/10.1016/j.lwt.2015.01.012]
[http://dx.doi.org/10.1016/j.lwt.2015.01.012]
[38]
Rathod NB, Elabed N, Punia S, Ozogul F, Kim SK, Rocha JM. Recent developments in polyphenol applications on human health: A review with current knowledge. Plants 2023; 12(6): 1217.
[http://dx.doi.org/10.3390/plants12061217] [PMID: 36986905]
[http://dx.doi.org/10.3390/plants12061217] [PMID: 36986905]
[39]
Diniz do Nascimento L, Moraes AAB, Costa KS, et al. Bioactive natural compounds and antioxidant activity of essential oils from spice plants: New findings and potential applications. Biomolecules 2020; 10(7): 988.
[http://dx.doi.org/10.3390/biom10070988] [PMID: 32630297]
[http://dx.doi.org/10.3390/biom10070988] [PMID: 32630297]
[40]
Vlčko T, Rathod NB, Kulawik P, Ozogul Y, Ozogul F. The impact of aromatic plant-derived bioactive compounds on seafood quality and safety. Advances in Food and Nutrition Research. Elsevier 2022; pp. 275-339.
[41]
Rathod NB, Ranveer RC, Benjakul S, et al. Recent developments of natural antimicrobials and antioxidants on fish and fishery food products. Compr Rev Food Sci Food Saf 2021; 20(4): 4182-210.
[http://dx.doi.org/10.1111/1541-4337.12787] [PMID: 34146459]
[http://dx.doi.org/10.1111/1541-4337.12787] [PMID: 34146459]
[42]
Bamisaye A, Eromosele CO, Dare EO, Akinloye OA, Idowu MA, Ighodaro OM, et al. Fabrication, characterization, antimicrobial, toxicity and potential drug-delivery studies of PEGylated Sesamum indicum oil based nanoemulsion system. Beni Suef Univ J Basic Appl Sci 2023; 12(1): 1-15.
[43]
Sharma M, Mann B, Pothuraju R, Sharma R, Kumar R. Physico-chemical characterization of ultrasound assisted clove oil-loaded nanoemulsion: As enhanced antimicrobial potential. Biotechnol Rep 2022; 34: e00720.
[http://dx.doi.org/10.1016/j.btre.2022.e00720] [PMID: 35686016]
[http://dx.doi.org/10.1016/j.btre.2022.e00720] [PMID: 35686016]
