Synergistic antibacterial effects of ultrasound and thyme essential oils nanoemulsion against Escherichia coli O157:H7

Essential oil nanoemulsions have been proven to have stronger antimicrobial effects compared to the essential oil alone or coarse emulsion. Sonoporation could be the promising candidate to trigger a synergistic effect with thyme essential oil nanoemulsion (TEON) and produce a more effective antibacterial efficacy. Therefore, in this study, the bactericidal effects of ultrasound (US) in combination with TEON treatments against Escherichia coli (E. coli) O157:H7 were investigated. The remarkable synergistic effects of US (20 kHz, 255 W/cm², 9 min) and TEON (0.375 mg/mL) treatments at 22 °C reduced E. coli O157:H7 populations by 7.42 ± 0.27 log CFU/mL.The morphological changes of cells exposed to different treatments were observed by scanning electron microscopy and transmission electron microscopy. The results showed that the synergistic effects of the ultrasound and TEON treatments altered the morphology and interior microstructure of organism cells. Laser scanning confocal microscopy (LSCM) images revealed that the combination treatments of ultrasound and TEON altered the permeability of cell membranes, and this affected the integrity of E. coli O157:H7 cells. This was further indicated by the high amounts of nucleic acids and proteins released from these cells following treatment.The results from this study illustrated the mechanisms of the synergistic effects of sonoporation and TEON treatments and provided valuable information for their potential in food pasteurization.     

Inactivation of foodborne pathogens based on synergistic effects of ultrasound and natural compounds during fresh produce washing

Ultrasound has potential to be used for disinfection, and its antimicrobial effectiveness can be enhanced in presence of natural compounds. In this study, we compared the antimicrobial effects of ultrasound at 20 kHz (US 20 kHz) or 1 MHz (US 1 MHz) in combination with carvacrol, citral, cinnamic acid, geraniol, gallic acid, lactic acid, or limonene against E. coli K12 and Listeria innocua at a constant power density in water. Compared to the cumulative effect of the individual treatments, the combined treatment of US 1 MHz and 10 mM citral generated >1.5 log CFU/mL additional inactivation of E. coli K12. Similarly, combined treatments of US 1 MHz and 2 mM carvacrol (30 min), US 20 kHz and 2 mM carvacrol, 10 mM citral, or 5 mM geraniol (15 min) generated >0.5–2.0 log CFU/mL additional inactivation in L. innocua. The synergistic effect of citral, as a presentative compound, and US 20 kHz treatment was determined to be a result of enhanced dispersion of insoluble citral droplets in combination with physical impact on bacterial membrane structures, whereas the inactivation by US 1 MHz was likely due to generation of oxidative stress within the bacteria. Combined ultrasound and citral treatments improved the bacterial inactivation in simulated wash water in presence of organic matter or during washing of inoculated blueberries but only additive antimicrobial effects were observed. Findings in this study will be useful to enhance fresh produce safety and shelf-life and design other alternative ultrasound based sanitation processes.     

Nondestructive monitoring,kinetics and antimicrobial properties of ultrasound technology applied for surface decontamination of bacterial foodborne pathogen in pork

In this study, electronic nose (E-nose) and Hyperspectral Imaging (HSI) was employed for nondestructive monitoring of ultrasound efficiency (20KHZ) in the inactivation of Salmonella Typhimurium, and Escherichia coli in inoculated pork samples treated for 10, 20 and 30 min.Weibull, and Log-linear model fitted well (R² ≥ 0.9) for both Salmonella Typhimurium, and Escherichia coli inactivation kinetics. The study also revealed that ultrasound has antimicrobial effects on the pathogens. For qualitative analysis, unsupervised (PCA) and supervised (LDA) chemometric algorithms were applied. PCA was used for successful sample clustering and LDA approach was used to construct statistical models for the classification of ultrasound treated and untreated samples. LDA showed classification accuracies of 99.26%,99.63%,99.70%, 99.43% for E-nose - S. Typhimurium, E-nose -E. coli, HSI - S. Typhimurium and HSI -E. coli respectively. PLSR quantitative models showed robust models for S. Typhimurium- (E-nose Rp² = 0.9375, RMSEP = 0.2107 log CFU/g and RPD = 9.7240 and (HSI Rp² = 0.9687 RMSEP = 0.1985 log CFU/g and RPD = 10.3217) and E. coli -(E-nose -Rp² = 0.9531, RMSEP = 0.2057 log CFU/g and RPD = 9.9604) and (HIS- Rp² = 0.9687, RMSEP = 0.2014 log CFU/g and RPD = 10.1731).This novel study shows the overall effectiveness of applying E-nose and HSI for in-situ and nondestructive detection, discrimination and quantification of bacterial foodborne pathogens during the application of food processing technologies like ultrasound for pathogen inactivation.     

Synergistic inactivation of bacteria based on a combination of low frequency,low-intensity ultrasound and a food grade antioxidant

This study evaluated a synergistic antimicrobial treatment using a combination of low frequency and a low-intensity ultrasound (LFU) and a food-grade antioxidant, propyl gallate (PG), against a model gram-positive (Listeria innocua) and the gram-negative bacteria (Escherichia coli O157:H7). Bacterial inactivation kinetic measurements were complemented by characterization of biophysical changes in liposomes, changes in bacterial membrane permeability, morphological changes in bacterial cells, and intracellular oxidative stress upon treatment with PG, LFU, and a combination of PG + LFU. Combination of PG + LFU significantly (>4 log CFU/mL, P < 0.05) enhanced the inactivation of both L. innocua and E. coli O157:H7 compared to PG or LFU treatment. As expected, L. innocua had a significantly higher resistance to inactivation compared to E. coli using a combination of PG + LFU. Biophysical measurements in liposomes, bacterial permeability measurements, and scanning electron microscope (SEM)-based morphological measurements show rapid interactions of PG with membranes. Upon extended treatment of cells with PG + LFU, a significant increase in membrane damage was observed compared to PG or LFU alone. A lack of change in the intracellular thiol content following the combined treatment and limited effectiveness of exogenously added antioxidants in attenuating the synergistic antimicrobial action demonstrated that oxidative stress was not a leading mechanism responsible for the synergistic inactivation by PG + LFU. Overall, the study illustrates synergistic inactivation of bacteria using a combination of PG + LFU based on enhanced membrane damage and its potential for applications in the food and environmental systems.     

Optimization and mechanism exploration for Escherichia coli transformed with plasmid pUC19 by the combination with ultrasound treatment and chemical method

As a basic technique of molecular cloning, bio-transformation has been successfully used in the fields of biomedicine and food processing. In this study, we established a transformation system of exogenous DNA into E. coli cells mediated by ultrasound. Under the optimal conditions (i.e. 35 °C, 40 W, 25 s, OD600 = 0.4–0.6) optimized by RSM, the transformation efficiency reached at 1.006 × 10⁷ CFU/μg DNA. The results of membrane permeability, macromolecular substance and cell structure analysis before and after ultrasound treatment showed that the damage of host cells induced by lower (40 W) ultrasound and shorter ultrasound time (25 s) was reversible, and the transformation efficiency and cell survival rate were not significantly affected under this condition. In brief, proper changes in cell membrane and cell wall were the basic conditions for host cells to uptake exogenous DNA, while, whether exogenous DNA could be replicated and expressed in cells depends on the viability of host cells.     

Study the impact of ultra-sonication and pulsed electric field on the quality of wheat plantlet juice through FTIR and SERS

Pulsed electric field (PEF) and Ultrasound (US) are commonly used in food processing. We investigated the combined impact of pulsed electric field (PEF) and ultrasound (US) on the wheat plantlet juice. When compared with the individual treatments, the highest values of total phenolics, total flavonoids, chlorophyll, ORAC assay, and DPPH activities were obtained using the combined (US + PEF) methods. The US + PEF significantly decreased the peroxidase and polyphenol oxidase activities from 0.87 to 0.27 Abs min⁻¹ and 0.031–0.016 Abs min⁻¹. Also, the synergistic application significantly lowered the yeast and mold (3.92 to 2.11 log CFU/mL), E. coli/Coliform (1.95 to 0.96 log CFU/mL), and aerobics (4.41 to 2.01 log CFU/mL). Furthermore, Fourier Transform Infrared (FT-IR) and surface-enhanced Raman spectroscopy (SERS) was used to analyzing juice quality. Gold nanoparticles (AuNPs) were used as the SERS substrates, which provided stronger Raman peaks for the samples treated with US + PEF methods. The FT-IR analysis showed significant enhancement of the nutritional molecules. The enhanced quality of wheat plantlet juice combined with lower yeast and mold suggests the suitability of integrated methods for further research and applications.     

Low-intensity ultrasound enhances the antimicrobial activity of neutral peptide TGH2 against Escherichia coli

In recent years, foodborne diseases caused by Escherichia coli are a major threat to the food industry and consumers. Antimicrobial peptides (AMPs) and ultrasound both have good inhibitory effects on E. coli. In this work, the mechanism of action and synergistic effect of an in silico predicted AMP, designated as TGH2 (AEFLREKLGDKCTDRHV), from the C-terminal sequence of Tegillarca granosa hemoglobin, combined with low-intensity ultrasound was explored. The minimal inhibitory concentration (MIC) of TGH2 on E. coli decreased by 4-fold to 31.25 μg/mL under 0.3 W/cm² ultrasound treatment, while the time kill curve analysis showed that low-intensity ultrasound combined with peptide TGH2 had an enhanced synergistic bactericidal effect after 0.5 h. The permeability on E. coli cell membrane increased progressively during combined treatment with peptide TGH2 and low-intensity ultrasound, resulting in the leakage of intracellular solutes, as shown by transmission electron microscopy (TEM). Structural analysis using circular dichroism (CD) revealed that peptide TGH2 has an α-helical structure, showing a slight untwisting effect under 0.3 W/cm² ultrasound treatment for 0.5 h. The findings here provide new insight into the potential application of ultrasound and AMPs combination in food preservation.     

Effects of static magnetic fields on Escherichia coli

This paper represents the study of the biological effects of static magnetic fields (SMFs) on Escherichia coli (E. coli). The bacterial strain E. coli was exposed to SMFs in order to test its viability (evaluated by the number of colony-forming units (CFU)). In this study, we measured the dependence of CFU on the duration of exposure, on the treatment temperature T and on the value of the magnetic field induction B. The results showed that the number of CFU decreased with longer exposure time and higher treatment temperature (from 25 °C to 40 °C), whereas multiple extreme values of number of CFU were obtained when the induction B changed. In order to explain the results, transmission electron microscopy (TEM) and scanning electron microscopy (SEM) were used to investigate the morphology of the cells. We found obvious cell surface damage when the cells were exposed to SMFs.     

Effect of ultrasound irradiation on bacterial internalization and bacteria-mediated gene transfer to cancer cells

The present study demonstrates that ultrasound irradiation can facilitate bacteria-mediated gene delivery (bactofection). Escherichia coli modified with avidin were employed as a vehicle for delivery of the green fluorescent protein (GFP) gene, a model heterologous gene, into the breast cancer cell line MCF-7. Avidin-mediated binding of E. coli to MCF-7 cells enhanced the internalization of E. coli by approximately 17%, irrespective of the use of ultrasound irradiation. Furthermore, the use of ultrasound irradiation increased the internalization by approximately 5%, irrespective of the presence of avidin on the E. coli cell surface. The percentages of GFP-expressing MCF-7 cells at 24 h after bactofection were below 0.5% and 2% for the case with only avidin-modification of E. coli cell surface and only ultrasound irradiation, respectively. However, combining avidin modification with the ultrasound treatment increased this value to 8%. Thus, the use of avidin-modified bacteria in conjunction with ultrasound irradiation has potential as an effective strategy for tumor-targeted bactofection.     

Synergistic effect of ultrasonic pre-treatment combined with UV irradiation for secondary effluent disinfection

The ultraviolet (UV) disinfection efficiency is often affected by suspended solids (SS). Given their high concentration or large particle size, SS can scatter UV light and provide shielding for bacteria. Thus, ultrasound is often employed as a pre-treatment process to improve UV disinfection. This work investigated the synergistic effect of ultrasound combined with UV for secondary effluent disinfection. Bench-scale experiments were conducted in using samples obtained from secondary sedimentation tanks. These tanks belonged to three wastewater treatment plants in Beijing that use different kinds of biological treatment methods. Several parameters may contribute to the changes in the efficiency of ultrasound and UV disinfection. Thus, the frequency and energy density of ultrasound, as well as the SS, were investigated. Results demonstrated that samples which have relatively higher SS concentrations or higher percentages of larger particles have less disinfection efficiency using UV disinfection alone. However, the presence of ultrasound could improve the disinfection efficiency because it has synergistic effect. Changes in the particle size distribution and SS concentration notably affected the efficiency of UV disinfection. The efficiency of Escherichia coli elimination can be decreased by 1.2 log units as the SS concentration increases from 16.9 mg/l to 25.4 mg/l at a UV energy density of 40 mJ/cm². UV disinfection alone reduced the E. coli population by 3.4 log units. However, the synergistic disinfection of ultrasound and UV could reach 5.4 log units during the reduction of E. coli at a 40 kHz frequency and an energy density of 2.64 kJ/l. The additional synergistic effect is 1.1 log units.     

Novel technique for treating grass carp (Ctenopharyngodon idella) by combining plasma functionalized liquids and Ultrasound: Effects on bacterial inactivation and quality attributes

A novel technique for treating grass carp by combining plasma functionalized liquids and ultrasound to inactivate bacteria was developed. The effects of the plasma functionalized liquids (PFL) including plasma functionalized water (PFW) and buffer (PFB) and their respective combination with ultrasound treatment (USPFW and USPFB) on the oxidative and physical qualities of grass carp were also investigated. Individual applications of PFW and PFB significantly reduced the populations of Escherichia coli and Shewanella putrefaciens in the range of 0.31–1.18 log CFU/g, compared with the control with a reduction of 0.18 log CFU/g, while combined treatments of USPFW and USPFB presented additional reductions of 0.05–0.65 log CFU/g, with potential synergy demonstrated for PFW and ultrasound. The treatment resulted in improved biomedical index and nutritional value of fatty acids and lipids, protein structural unfolding, increased lipid oxidation and protein degradation with values within the acceptable limits, and the combined treatment was more effective for retarding the hardness reduction in grass carp, while the colour change was also significantly affected, resulting in increased whiteness. The results indicated that the combined treatments may be a promising approach to improving the quality of seafood products.     

Combination of therapeutic ultrasound with antibiotics interfere with the growth of bacterial culture that colonizes skin ulcers: An in-vitro study

Staphylococcus aureus and Escherichia coli are among the major bacterial species that colonize skin ulcers. Therapeutic ultrasound (TUS) produces biophysical effects that are relevant to wound healing; however, its application over a contaminated injury is not evidence-based. The objective of this research was to analyze the effect of TUS on in vitro-isolated S. aureus and E. coli, including the combination of ultrasound and antibiotics, in order to assess their antibiotic action on bacterial susceptibility. For the experiments, the bacterial strains were suspended in saline, then diluted (10⁴ CFU/mL) for irradiation (at 1 and 3 MHz, 0.5 and 0.8 W/cm² for 0 and 15 min) and the combination treatment of ultrasonication and antibiotics was administered by adding nalidixic acid (S. aureus) and tetracycline (E. coli) at concentrations equivalent to 50% of the minimum inhibitory concentration (MIC). The experiments were carried out in duplicate with six repetitions. The suspensions were inoculated on to Petri plates and incubated at 37 °C and the colony forming units (CFUs) were counted after 24 h. The results were subjected to the Shapiro–Wilk normality test, followed by parametric ANOVA and Tukey’s post hoc test at a significance level of 1%. The results demonstrated that the action of TUS at 1 MHz inhibited bacterial growth while at 3 MHz, bacterial growth was observed in both species. However, the synergistic combination of ultrasound and antibiotics was able to inhibit the growth of both bacteria completely after 15 min of ultrasonication. The results suggest that the action of ultrasound on S. aureus and E. coli are dependent on the oscillation frequency as well as the intensity and time of application. The combination of ultrasound with antibiotics was able to inhibit bacterial growth fully at all frequencies and doses in both species.     

Sonophotocatalytic inactivation of E. coli using ZnO nanofluids and its mechanism

The present study evaluated inactivation efficiency of a sonophotocatalytic process using ZnO nanofluids including ultrasonic parameters such as power density, frequency and time. The result showed that inactivation efficiency was increased by 20% when ultrasonic irradiation was combined with photocatalytic process in the presence of natural light. Comparison of inactivation efficiency in photocatalytic, ultrasonic and sonocatalytic processes using Escherichia coli as a model bacteria identified that inactivation efficiencies are shown in the following order: ultrasonic irradiation < sonocatalysis < photocatalysis < sonophotocatalysis. Furthermore, inactivation mechanism of sonophotocatalysis was proposed. Studies of reactive oxygen species (ROS) and zinc ions (Zn²⁺) release evaluation revealed that ROS play a key role in bacterial inactivation rather than Zn²⁺. Permeability of outer membrane (OM) and inner membrane (IM) of E. coli bacterial cells were studied and exhibited that sonophotocatalysis increased the permeability of OM and IM significantly. The enhanced bacterial inactivation effect in sonophotocatalytic process contributed to acoustic cavitation, sonocatalysis of ZnO and sonoporation phenomenon.     

Sonozonation: Enhancing the antimicrobial efficiency of aqueous ozone washing techniques on cherry tomato

Ultrasound requires high power and longer treatment times to inactivate microorganisms when compared to ultrasound combined with other technologies. Also, the antimicrobial efficiency of aqueous ozone increases with an increase in its concentration and exposure time, but with a detrimental effect on the quality of the treated food. In this study, the effect of aqueous ozone at low concentration, multi-mode frequency irradiation and their combination on microbial safety and nutritional quality of cherry tomato was investigated. Individual washing with aqueous ozone and mono-mode frequency irradiation resulted in <1 log CFU/g reduction in the spoilage microorganisms, while dual-mode frequency irradiation (DMFI) resulted in higher microbial reduction (1.3–2.6 1 log CFU/g). The combined system (20/40 kHz + aqueous ozone) on the other hand, resulted in >3 log CFU/g microbial reduction. The application of DMFI enhanced the antimicrobial efficiency of aqueous ozone without any detrimental effect on the physicochemical properties (except the firmness), bioactive compounds, and antioxidants of the cherry tomato during 21 days refrigerated storage. The result obtained indicates the promising substitute to the single washing technique for microbial safety as well as preserving the nutritional quality and enhancing the shelf life of cherry tomato.     

Effect of ultrasound and chlorine dioxide on Salmonella Typhimurium and Escherichia coli inactivation in poultry chiller tank water

This study evaluated the application of ultrasound alone or combined with chlorine dioxide (ClO₂) for Salmonella Typhimurium and Escherichia coli inactivation in poultry processing chiller tank water. A Full Factorial Design (FFD) 2² was conducted for each microorganism to evaluate the effect of ultrasound exposure time (x₁: 1 to 9 min; fixed: 37 kHz; 330 W; 25 °C) using a bath, and ClO₂ concentration (x₂: 1 to 17 mg L⁻¹) on microorganism count expressed in log CFU mL⁻¹ in distilled water. Variable x₂ had a negative effect on Salmonella Typhimurium (-5.09) and Escherichia coli (-2.00) count, improving the inactivation; while a x₁ increase present no inactivation improvement, explaining the use of x₁ lower level (1 min) and x₂ higher level (17 mg L⁻¹). The best condition for microorganism inactivation based on FFD was evaluated in chiller tank water (with organic matter) at 25, 16, and 4 °C; x₁ was kept (1 min), however x₂ was adjusted to obtain the same residual free chlorine (2.38 mg L⁻¹) considering the ClO₂ consumption by organic matter, achieving the value of 30 mg L⁻¹. An inactivation of 49% and 31% were observed for Salmonella Typhimurium and Escherichia coli. When ultrasound was replaced by a simple agitation in the presence of ClO₂, there was no inactivation for both microorganisms. Moreover, at poultry carcass pre-chilling (16 °C) and chilling (4 °C) conditions, the synergism of ultrasound combined with ClO₂ was more pronounced, with microorganisms’ reductions up to 100%.     

Disinfection effect of a continuous-flow ultrasound/ultraviolet baffled reactor at a pilot scale

An ultrasound/ultraviolet (US/UV) baffled reactor was developed to fill the gap in ultraviolet (UV) disinfection associated with disinfection efficiency. According to the previously selected operational condition, a continuous-flow US/UV baffled reactor was continuously operated in a wastewater treatment plant at a pilot scale for nearly three months, and the disinfection influent and effluent were analyzed, including fecal coliforms, Escherichia coli, and fecal streptococci. The US/UV baffled reactor could guarantee a high effluent disinfection performance in terms of fecal coliforms removal even with the fluctuation of the secondary effluent. All the disinfected effluents satisfied the requirement of the “Pollutants Discharge Standard of Municipal Wastewater Treatment Plant in China” (fecal coliforms below 1000 CFU/L for class 1A), and 87% of the tested fecal coliforms concentration in the disinfected effluent was below 100 CFU/L, nearly eliminating all fecal coliforms. Further analysis of the E. coli and fecal streptococci showed the broad disinfection ability and high disinfection efficiency of the US/UV baffled reactor. The flexibility of the specific energy consumption for the disinfection system depends on the water quality.     

Synergetic effects of ultrasound and slightly acidic electrolyzed water against Staphylococcus aureus evaluated by flow cytometry and electron microscopy

This study evaluated the synergetic effects of ultrasound and slightly acidic electrolyzed water (SAEW) on the inactivation of Staphylococcus aureus using flow cytometry and electron microscopy. The individual ultrasound treatment for 10 min only resulted in 0.36 log CFU/mL reductions of S. aureus, while the SAEW treatment alone for 10 min resulted in 3.06 log CFU/mL reductions. The log reductions caused by combined treatment were enhanced to 3.68 log CFU/mL, which were greater than the sum of individual treatments. This phenomenon was referred to as synergistic effects. FCM analysis distinguished live and dead cells as well as revealed dynamic changes in the physiological states of S. aureus after different treatments. The combined treatment greatly reduced the number of viable but nonculturable (VBNC) bacteria to 0.07%; in contrast, a single ultrasound treatment for 10 min induced the formation of VBNC cells to 45.75%. Scanning and transmission electron microscopy analysis revealed that greater damage to the appearance and ultrastructure of S. aureus were achieved after combined ultrasound-SAEW treatment compared to either treatment alone. These results indicated that combining ultrasound with SAEW is a promising sterilization technology with potential uses for environmental remediation and food preservation.     

Synergistic bactericidal effect of ultrasound combined with citral nanoemulsion on Salmonella and its application in the preservation of purple kale

In this study, a novel citral nanoemulsion (CLNE) was prepared by ultrasonic emulsification. The synergistic antibacterial mechanism of ultrasound combined with CLNE against Salmonella Typhimurium and the effect on the physicochemical properties of purple kale were investigated. The results showed that the combined treatment showed obviously inactivate effect of S. Typhimurium. Treatment with 0.3 mg/mL CLNE combined with US (20 kHz, 253 W/cm²) for 8 min reduced S. Typhimurium populations in phosphate-buffered saline (PBS) by 9.05 log CFU/mL. Confocal laser scanning microscopy (CLSM), flow cytometry (FCM), protein and nucleic acid release assays showed that the US combination CLNE disrupt the integrity of S. Typhimurium membranes. Reactive oxygen species (ROS) and malondialdehyde (MDA) detection indicated that US+CLNE exacerbated oxidative stress and lipid peroxidation in cell membranes. The morphological changes of cells after different treatments by field emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM) illustrated that the synergistic effect of US+CLNE treatment changed the morphology and internal microstructure of the bacteriophage cells. Application of US+CLNE on purple kale leaves for 6 min significantly (P < 0.05) reduced the number of S. Typhimurium, but no changes in the physicochemical properties of the leaves were detected. This study elucidates the synergistic antibacterial mechanism of ultrasound combined with CLNE and provides a theoretical basis for its application in food sterilization.     

Ultrasonic-assisted preparation of eucalyptus oil nanoemulsion: Process optimization,in vitro digestive stability,and anti-Escherichia coli activity

Eucalyptus oil (EO) is a natural and effective antimicrobial agent; however, it has disadvantages such as poor water solubility and instability. The aim of this study was to investigate the effect of process vessels and preparation process parameters on the particle size of the emulsion droplets using ultrasonic technique and response surface methodology to prepare eucalyptus oil nanoemulsion (EONE). The optimal sonication process parameters in conical centrifuge tubes were confirmed: sonication distance of 0.9 cm, sonication amplitude of 18%, and sonication time of 2 min. Under these conditions, the particle size of EONE was 18.96 ± 4.66 nm, the polydispersity index was 0.39 ± 0.09, and the zeta potential was −31.17 ± 2.15 mV. In addition, the changes in particle size, potential, micromorphology, and anti-Escherichia coli activity of EONE during digestion were investigated by in vitro simulated digestion. The emulsion was stable in simulated salivary fluid, tended to aggregate in simulated gastric fluid, and increased in particle size and potential value in simulated intestinal fluid. EONE showed higher anti-E. coli activity than EO by simulated digestion. These results provide a useful reference for the in vivo antimicrobial application of the essential oil.