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.     

The combination of ultrasound and chlorogenic acid to inactivate Staphylococcus aureus under planktonic,biofilm, and food systems

This study aimed to investigate the mechanism of different treatments, namely, ultrasound (US), chlorogenic acid (CA), and ultrasound combined with chlorogenic acid (US plus CA) on the inactivation of Staphylococcus aureus planktonic and biofilm cells. Results showed that the combined treatment of US and CA exhibited remarkable synergistic antibacterial and antibiofilm effects. Scanning electron microscopy images indicated that the combined treatment of US and CA caused the most serious damage to the cell morphology. Confocal laser scanning microscopy images revealed that the combined treatment led to sharp increase and severe damage to the permeability of the cell membrane, causing the release of ATP and nucleic acids and decreasing the exopolysaccharide contents in S. aureus biofilm. Finally, the combined treatment of US plus 1% CA for 60 min inactivated S. aureus cells by 1.13 lg CFU/g on mutton. Thus, the combined treatment of US and CA had synergistic effect against S. aureus under planktonic, biofilm, and food systems.     

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.     

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.     

Ultrasonic irradiation enhanced the efficacy of antimicrobial photodynamic therapy against methicillin-resistant Staphylococcus aureus biofilm

Antimicrobial photodynamic therapy (aPDT) is a non-pharmacological antimicrobial regimen based on light, photosensitizer and oxygen. It has become a potential method to inactivate multidrug-resistant bacteria. However, limited by the delivery of photosensitizer (PS) in biofilm, eradicating biofilm-associated infections by aPDT remains challenging. This study aimed to explore the feasibility of combining ultrasonic irradiation with aPDT to enhance the efficacy of aPDT against methicillin-resistant staphylococcus aureus (MRSA) biofilm. A cationic benzylidene cyclopentanone photosensitizer with much higher selectivity to bacterial cells than mammalian cells were applied at the concentration of 10 μM. 532 nm laser (40 mW/cm², 10 min) and 1 MHz ultrasound (500 mW/cm², 10 min, simultaneously with aPDT) were employed against MRSA biofilms in vitro. In addition to combined with ultrasonic irradiation and aPDT, MRSA biofilms were treated with laser irradiation only, photosensitizer only, ultrasonic irradiation only, ultrasonic irradiation and photosensitizer, and aPDT respectively. The antibacterial efficacy was determined by XTT assay, and the penetration depth of PS in biofilm was observed using a photoluminescence spectrometer and a confocal laser scanning microscopy (CLSM). In addition, the viability of human dermal fibroblasts (WS-1 cells) after the same treatments mentioned above and the uptake of P3 by WS-1 cells after ultrasonic irradiation were detected by CCK-8 and CLSM in vitro. Results showed that the percent decrease in metabolic activity resulting from the US + aPDT group (75.76%) was higher than the sum of the aPDT group (44.14%) and the US group (9.88%), suggesting synergistic effects. Meanwhile, the diffusion of PS in the biofilm of MRSA was significantly increased by 1 MHz ultrasonic irradiation. Ultrasonic irradiation neither induced the PS uptake by WS-1 cells nor reduced the viability of WS-1 cells. These results suggested that 1 MHz ultrasonic irradiation significantly enhanced the efficacy of aPDT against MRSA biofilm by increasing the penetration depth of PS. In addition, the antibacterial efficacy of aPDT can be enhanced by ultrasonic irradiation, the US + aPDT treatment demonstrated encouraging in vivo antibacterial efficacy (1.73 log₁₀ CFU/mL reduction). In conclusion, the combination of aPDT and 1 MHz ultrasound is a potential and promising strategy to eradicate biofilm-associated infections of MRSA.     

Antibacterial mechanism of ultrasound against Escherichia coli: Alterations in membrane microstructures and properties

This study was aimed at providing new insights on the response of bacterial cell membranes to ultrasound exposure. Escherichia coli (E. coli) O157:H7 cells were exposed to different ultrasound treatments (power intensities of 64, 191, 372, and 573 W/cm², frequency of 20 kHz, pulsed mode of 2 sec: 2 sec) and the dynamic changes in cell viability within 27 min were assessed. With an increase in ultrasonic intensity and prolonged duration, a 0.76–3.52 log CFU/mL reduction in E. coli populations was attained. The alterations in the sensitivity of ultrasound-treated cells to antimicrobial compounds were evaluated by exposure to thyme essential oil nanoemulsion (TEON). The treatment reduced the E. coli population by 2.16–7.10 log CFU/mL, indicating the effects of ultrasonic field on facilitating the antibacterial efficacy of TEON. Ultrasonic-treated E. coli cells also displayed remarkable morphological and ultrastructural damages with destroyed membrane integrity and misshaped cell structures, which was observed by electron microscopy analysis. Significant increase in outer and inner membrane permeability, along with the cytoplasmic leakage and membrane depolarization were assessed utilizing spectrophotometry. For the first time, significant reduction in the membrane fluidity in response to ultrasound exposure were investigated. Additional efforts in exploring the effect of ultrasonic field on some bacterial membrane compositions were performed with infrared spectroscopy. In this study, multiple lines of evidence effectively served to elucidate the alterations on cellular membrane structure and property during exposure to sonication that could extend our understanding of the antimicrobial molecular mechanisms of ultrasound.     

Use of aqueous ozone rinsing to improve the disinfection efficacy and shorten the processing time of ultrasound-assisted washing of fresh produce

In minimal processing industry, chlorine is widely used in the disinfection process and ultrasound (US) increases the disinfection efficacy of chlorine and reduces the cross-contamination incidence during washing. Tap water (TW), which has no disinfection effect, is generally used to rinse off sanitizer residues on the surface of disinfected fresh-cut vegetables. In this study, aqueous ozone (AO), a low-cost and residue-free sanitizer, was used to replace TW rinsing in combination with US (28 kHz)–chlorine (free chlorine [FC] at 10 ppm, a concentration recommended for industrial use) for the disinfection of fresh-cut lettuce as a model. US–chlorine (40 s) + 2.0 ppm AO (60 s) treatment resulted in browning spots on lettuce surface at the end of storage. In contrast, US–chlorine (40 s) + 1.0 ppm AO (60 s) did not lead to deterioration of the sensory quality (sensory crispness, color, and flavor) and a change in total color difference, and the activities of browning-related enzymes were significantly lower. Moreover, US–chlorine (40 s) + 1.0 ppm of AO (60 s) treatment led to significantly lower counts of Escherichia coli O157:H7, Salmonella Typhimurium, aerobic mesophilic (AMC), and molds and yeasts (M&Y) on days 0–7 than US–chlorine (60 s) + TW (60 s) and single 1.0 ppm AO (120 s) treatments, suggesting that AO provided an additional disinfection effect over TW, while reducing the overall processing time by 20 s. Cell membrane permeability analysis (alkaline phosphatase, protein, nucleotide, and adenosine triphosphate leakage) showed that the combination with 1.0 ppm AO caused more severe cell membrane damage in E. coli O157:H7 and S. Typhimurium, explaining the higher disinfection efficacy. 16S rRNA sequencing revealed that following US–chlorine (40 s) + 1.0 ppm of AO (60 s) treatment, Massilia and Acinetobacter had higher relative abundances (RAs) on day 7 than after US–chlorine (60 s) + TW (60 s) treatment, whereas the RAs of Escherichia–Shigella was significantly lower, indicating that the former treatment has a superior capacity in maintaining a stable microbial composition. This explains from an ecological point of view why US–chlorine (40 s) + 1.0 ppm of AO (60 s) led to the lowest AMC and M&Y counts during storage. The study results provide evidence that AO has potential as an alternative to TW rinsing to increase the disinfection efficacy of US–chlorine.     

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.     

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.     

The modification of pomegranate polyphenol with ultrasound improves mechanical,antioxidant, and antibacterial properties of tuna skin collagen-chitosan film

To produce an edible film with high mechanical and physicochemical properties, Tuna skin collagen-chitosan (TSC-CTS) composite films were prepared by incorporating ultrasound (UT) and pomegranate polyphenols including gallic acid (GA), tannic acid (TA), and ellagic acid (EA), respectively. The tensile strength and the DPPH scavenging activity of the GA-UT-TSC-CTS film (ultrasound frequency of 28 ± 0.5 kHz, power of 100 W/L, sweep frequency cycle of 100 ms, duty ratio of 77% and time of 10 min; GA concentration of 1.0 g/L and reaction time of 10 min) were increased by 47.03% and 24.16 folds, respectively compared to the control (TSC-CTS film). Meanwhile, light transmittance and water vapor permeability of the GA-UT-TSC-CTS film were decreased by 29.26% and 15.70%, respectively. These positive modification results were attributed to the altered structure during the film formation process, which were verified by Fourier transform infrared spectroscopy (FTIR), circular dichroism (CD), X-ray diffraction (XRD), and thermogravimetry results. Moreover, the GA-UT-TSC-CTS film possessed moderate thermal stability and color indexes and improved antibacterial activity. The antibacterial effect of the film against Bacillus subtilis was the highest, followed by Escherichia coli, Listeria monocytogenes, and Staphylococcus aureus. Overall, the combination modification of gallic acid and ultrasound was an efficient modification method to improve the mechanical, antioxidant, and antibacterial properties of edible TSC-CTS films.     

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%.     

Sonophotocatalytic degradation of trypan blue and vesuvine dyes in the presence of blue light active photocatalyst of Ag3PO4/Bi2S3-HKUST-1-MOF: Central composite optimization and synergistic effect study

An efficient simultaneous sonophotocatalytic degradation of trypan blue (TB) and vesuvine (VS) using Ag₃PO₄/Bi₂S₃-HKUST-1-MOF as a novel visible light active photocatalyst was carried out successfully in a continuous flow-loop reactor equipped to blue LED light. Ag₃PO₄/Bi₂S₃-HKUST-1-MOF with activation ability under blue light illumination was synthesized and characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), energy dispersive X-ray (EDX), photoluminescence (PL) and diffuse reflectance spectra (DRS). The effect of operational parameters such as the initial TB and VS concentration (5–45 mg/L), flow rate (30–110 mL/min), irradiation and sonication time (10–30 min), pH (3–11) and photocatalyst dosage (0.15–0.35 g/L) has been investigated and optimized using central composite design (CCD) combined with desirability function (DF). Maximum sonophotodegradation percentage (98.44% and 99.36% for TB and VS, respectively) was found at optimum condition set as: 25 mg/L of each dye, 70 mL/min of solution flow rate, 25 min of irradiation and sonication time, pH 6 and 0.25 g/L of photocatalyst dosage. At optimum conditions, synergistic index value was obtained 2.53 that indicated the hybrid systems including ultrasound irradiation and photocatalysis have higher efficiency compared with sum of the individual processes.     

Synergistic antibacterial and antibiofilm effects of ultrasound and MEL-A against methicillin-resistant Staphylococcus aureus

Methicillin-resistant Staphylococcus aureus (MRSA) is drug-resistant and biofilm-forming pathogenic bacteria with severe morbidity and mortality, and has been continuously detected in food products in recent years. Mannosylerythritol lipids (MELs) are novel biosurfactants and perform antibacterial property against gram-positive bacteria. Ultrasound has been applied into food sterilization as non-thermal techniques and has advantage of maintaining food nutrition and flavor over heat pasteurization. In this work, the synergistic treatment of ultrasound and MEL-A was used to combat planktonic cells and biofilm of MRSA. As a result, the combined treatment has exhibited remarkable antibacterial effect proved by enumeration of viable microbes. Furthermore, flow cytometry, scanning electron microscopy and transmission electron microscopy revealed ultrasound has enhanced the inhibitory effect of MEL-A through exacerbating cell membrane damage. On the other hand, the collaborating working modes to eradicate MRSA biofilm were disturbing cell adhesion to surface by MEL-A and destructing mature biofilm mechanically by ultrasound, reaching to over 90% of clearance rate. The findings of this study illustrated the synergistic antimicrobial mechanism of ultrasound and MEL-A treatments, and offered theoretical basis for their potential applications in food preservation.     

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.     

Mechanical damage and thermal effect induced by ultrasonic treatment in olive leaf tissue. Impact on polyphenols recovery

The influence of ultrasound treatment (US) on cellular damage of olive leaf tissue was studied. Mechanical damage and thermal effect of US were characterized. The level of tissue damage was defined by the diffusivity disintegration index Z_D based on the diffusivity of solutes extracted from olive leaves differently treated. The Arrhenius form using the temperature dependences of the thermal treatment time within the temperature interval 20–90 °C was observed for the thermal process. The corresponding activation energy ΔUT was estimated as 57 kJ/mol. The temperature dependences of electrical conductivity were measured for extracts of intact and maximally treated olive leaves. Then the diffusivity disintegration index Z_D and total phenolic compounds recovery for three studied US powers were calculated (100, 200, and 400 W). The results evidenced that the mechanically stimulated damage in olive leaf tissue can occur even at a low US power of 100 W if treatment time is long enough (t = 3.5 h). The US treatment noticeably accelerated the diffusion process mechanically in addition to its thermal effect. Trials in aqueous solution revealed the dependence of polyphenols extraction on damage level with respect to the US power applied.     

Structural and permeability sensitivity of cells to low intensity ultrasound: Infrared and fluorescence evidence in vitro

This work is focused on the in vitro study of the effects induced by medical ultrasound (US) in murine fibroblast cells (NIH-3T3) at a low-intensity of exposure (spatial peak temporal average intensity I_ta < 0.1 W cm⁻²). Conventional 1 MHz and 3 MHz US devices of therapeutic relevance were employed with varying intensity and exposure time parameters. In this framework, upon cells exposure to US, structural changes at the molecular level were evaluated by infrared spectroscopy; alterations in plasma membrane permeability were monitored in terms of uptake efficiency of small cell-impermeable model drug molecules, as measured by fluorescence microscopy and flow cytometry. The results were related to the cell viability and combined with the statistical PCA analysis, confirming that NIH-3T3 cells are sensitive to therapeutic US, mainly at 1 MHz, with time-dependent increases in both efficiency of uptake, recovery of wild-type membrane permeability, and the size of molecules entering 3T3. On the contrary, the exposures from US equipment at 3 MHz show uptakes comparable with untreated samples.     

Bioactive compounds and antibacterial activities in crystallized honey liquefied with ultrasound

The effect of ultrasound on the crystal size, phenols, flavonoids, Maillard products and antibacterial activity of crystallized honeys was studied. Three multifloral honeys (M), one monofloral (MO) and one honeydew (HD) honey were used. Ultrasound was performed at 42 kHz for different times (0, 5, 10 and 15 min). The antibacterial activities were tested against Salmonella typhimurium, Bacillus subtilis, Pseudomonas aeruginosa, Listeria monocytogenes, Staphylococcus aureus and Escherichia coli. In all honeys, the parameters analyzed had significant differences ((P < 0.05)). After 15 min of ultrasound the HD had increments of 44 mg of gallic acid/100 g of honey in phenols, and some M showed increase in flavonoids (5.64 mg of quercitin /100 g of honey) and improvement in inhibition against Salmonella typhimurium was 13.1%. In some honeys the correlation between phenols or flavonoids and antibacterial activity were significant ((P < 0.05)). No correlation was found between Maillard products and antibacterial activity. The ultrasound treatment effect on the crystal size, phenols, flavonoid, Maillard products, and antibacterial activity of crystallized honeys were different in each honey.     

Combination of ozone and ultrasonic-assisted aerosolization sanitizer as a sanitizing process to disinfect fresh-cut lettuce

Reduction of sanitizer dosage and development of non-immersion disinfection methods have become major focuses of research. Here, we examined the disinfection efficacy of combining gaseous ozone (4 and 8 ppm) with aerosolized oxidizing sanitizer [sodium hypochlorite (SH, 100 and 200 ppm)] and aerosolized organic acid [acetic acid (AA, 1% and 2%) and lactic acid (LA, 1% and 2%)]. Notably, 1% AA and 4 ppm gaseous ozone were ineffective for disinfecting Salmonella Typhimurium, and treatment with 1% AA + 8 ppm ozone caused browning of lettuce leaves and stimulated increases in aerobic mesophilic count (AMC), aerobic psychrotrophic count (APC), S. Typhimurium, and Escherichia coli O157:H7. Treatment with 2% LA + 8 ppm ozone resulted in the lowest S. Typhimurium, E. coli O157:H7, Listeria monocytogenes, AMC, APC, and molds and yeasts during storage (0–7 days at 4 °C). Quality analysis indicates that LA + 8 ppm ozone and SH + 8 ppm ozone did not negatively affect L*, a*, b*, polyphenolic content, weight loss, and sensory properties; however, the levels of two individual phenolic compounds (3,4-dihydroxybenzoic acid and vanillin), responsible for phenylpropanoid synthesis, were significantly increased after treatment with 2% LA + 8 ppm ozone. These findings provided insights into the use of LA combined with gaseous ozone for application in disinfecting fresh produce.     

Developing ultrasound-assisted hot-air and infrared drying technology for sweet potatoes

The influence of ultrasound (US) pretreatments combined with infrared (IRD) and hot-air (HAD) drying on drying kinetics, mathematical modeling, bioactive compounds (antioxidant activities, Vitamin C, phenolics, and flavonoid contents), qualitative properties (β-carotene, total carotenoids, color indexes, textural profile), enzyme inactivation, and exergetic analysis of sweet potatoes. The US pretreatment at 40 kHz combined with IRD and HAD (70 °C) significantly lessened the drying time and water contents. Besides, it did not affect the sweet potato's bioactive components and other quality-related attributes. The samples’ activation energy (E_a) ranged from 17.60 to 29.86 kJ/mol for both dryers, with R² (0.999–0.9809). Control samples had the highest specific energy consumption (SEC) due to the extended drying period, whereas ultrasound (40 kHz) treated samples had the lowest SEC during HAD and IRD at 80 °C. The thermodynamic parameters indicated that increasing the drying temperature lowers the enthalpy and Gibbs free energy, while entropy resulted in negative values. HAD had better textural qualities (hardness and resilience). The US pretreatments followed by HAD or IRD may lead to an energy-efficient method with acceptable quality maintenance.     

A novel approach to quantitative ultrasonic naked gene delivery and its non-invasive assessment

The purpose of this study was to investigate practical, safe, easy-to-use, non-cytotoxic, and reliable parameters to apply to an ultrasound (US) naked gene therapy system. The ultrasound pressure at the point of cell exposure was measured using a calibrated hydrophone and the intensity calculated. An acoustic power meter calibrated using a hydrophone was used to measure the power of the transducer. Four cell types were exposed to US with different exposure times and intensities. Fluorescent microscopy, spectrophotometry, scanning electron microscope, laser scanning confocal microscopy, flow cytometry and histogram analysis were used to evaluate the results of the study. The plasmid of green fluorescent protein (GFP) served as the reporter gene. The energy accumulation E in US gene delivery for 90% cell survival was defined as the optimal parameters (E = 3.56 ± 0.06), and at 80% cell survival was defined as the damage threshold (E = 59.67 ± 3.54). US safely delivered GFP into S180 cells (35.1 kHz) at these optimal parameters without obvious damage or cytotoxity in vitro. Exposed cell function was proved normal in vivo. The transfection rate was 35.83 ± 2.53% (n = 6) in viable cells, corresponding to 90.17 ± 1.47% (n = 6) cell viability. The intensity of GFP expression showed a higher fluorescent peak in the group of adeno-associated virus GFP vector (AVV-GFP) than in the control group (P < 0.001). The effect of US gene delivery and cell viability correlated as a fifth order polynomial with US intensity and exposure time. With optimal parameters, US can safely deliver naked a gene into a cell without damage to cell function. Both optimal uptake and expression of gene depend on the energy E at 90% cell survival. E can be applied as a control factor for bioeffects when combined with other parameters. Stable caviation results in optimal parameters for gene delivery and the transient caviation may cause cell damage, which will bring about a sharp rise of permeabilization. The results may be applied to the development of a novel clinical gene therapeutic system.