Effect of ultrasound combined with ultraviolet treatment on microbial inactivation and quality properties of mango juice

This work explored the effect of ultraviolet-assisted ultrasound (US-UV) as an emerging non-thermal sterilization technology on mango juice in aspects of microbial growth and quality changes. The juice in the ice bath was subjected to US-UV treatment at different US powers (0–600 W) and times (0–40 min), and no pathogen bacteria could be detected after treatment, while the physicochemical features (particle size, suspension stability, color, content of total polyphenols, carotenoids, sugar, reducing sugar and protein) and antioxidant ability of treated juice was preserved or improved to some extent. Based on these results, we further validated its positive effects on the nutritional value (content of ascorbic acid and soluble dietary fiber, antioxidant ability) and quality parameters (titratable acid, sugar acidity, total soluble solids, rheological behavior, metal elements) of mango juice treated at the optimal US parameter (10 min, 600 W); Not only the inactivation of polyphenol oxidation enzyme, peroxidase and pectin methylesterase was achieved but also the treated juice has a significant different volatile profile compared with the fresh juice, which might offer the better color, texture, and smell. Importantly, through the HPLC-MSD-Trap-XCT (phenols) and UPLC-Q Exactive Orbitrap-MS (carotenoids) study, the US-UV treatment will not cause difference on compounds composition, but it was responsible for changes in content of individual compounds, especially the all-trans-β-carotene, became the main component of carotenoids in processed mango juice (increased from 43.72% to 75.15%, relative content), and the oxygenated carotenoids (xanthophylls) are highly sensitive to the US (reduced from 50.96% to 4.85%) while the carotenes show a strong resistance to the US (increased 49.04% to 95.15%). Thus, the overall safety and quality of mango juice were enhanced while the sensory characteristics remained stable, suggesting that this non-thermal combination sterilization processing may successfully be implemented in the commercial processing of mango juice.     

Synergism between ultrasonic pretreatment and white rot fungal enzymes on biodegradation of wheat chaff

Lignocellulosic biomass samples (wheat chaff) were pretreated by ultrasound (US) (40 kHz/0.5 W cm⁻²/10 min and 400 kHz/0.5 W cm⁻²/10 min applied sequentially) prior to digestion by enzyme extracts obtained from fermentation of the biomass with white rot fungi (Phanerochaete chrysosporium or Trametes sp.). The accessibility of the cellulosic components in wheat chaff was increased, as demonstrated by the increased concentration of sugars produced by exposure to the ultrasound treatment prior to enzyme addition. Pretreatment with ultrasound increased the concentration of lignin degradation products (guaiacol and syringol) obtained from wheat chaff after enzyme addition. In vitro digestibility of wheat chaff was also enhanced by the ultrasonics pretreatment in combination with treatment with enzyme extracts. Degradation was enhanced with the use of a mixture of the enzyme extracts compared to that for a single enzyme extract.     

The effect of high-power ultrasound on the rheological properties of strawberry pulp

This study investigated the effects of high-power ultrasound (HPU, 0–45 °C, 242–968 W/cm², 2–16 min) on the rheological properties of strawberry pulp. Following the HPU treatment, the strawberry pulp exhibited an increase in apparent viscosity, storage modulus (G′), and loss modulus (G″). The water-soluble pectin (WSP), pectin methylesterase (PME) activity, and free calcium ions (Ca²⁺) of the strawberry pulp after HPU treatment were investigated to determine a possible reason for this phenomenon. HPU caused a significant decrease in the degree of esterification (DE), molecular weight (Mw), and particle size of strawberry WSP, but no significant changes were evident in the galacturonic acid (GalA) content and the zeta (ζ)-potential (P > 0.05), resulting in decrease in the apparent viscosity. Moreover, the largest reduction of PME activity was 22.6% after HPU treatment at 605 W/cm² and 45 °C for 16 min, indicating that the PME was resistant to the HPU treatments. The free Ca²⁺ content in the strawberry pulp was significantly decreased after exposure to HPU (P < 0.05). The maximal reduction of 52.01% in the free Ca²⁺ was achieved at 605 W/cm² and 45 °C for 16 min. The overall results indicated that the high residual activity (RA) of PME after HPU might induce the low esterification of WSP, while HPU promoted the interaction of free Ca²⁺ and low-methylated pectin, to form the network structure of Ca²⁺-low-methylated pectin, resulting in an increase in viscosity in the complex strawberry system.     

A chemometric approach to evaluate the effects of probe-type ultrasonication on the enzyme inactivation and quality attributes of fresh amla juice

The enzymatic browning induced in amla juice due to the high activity of polyphenol oxidase (PPO) and peroxidase (POD) is one of the critical issues faced by the industry. The present study assessed the suitability of non-thermal, high-intensity ultrasound (US) on the inactivation of PPO and POD in fresh Indian Gooseberry juice. Ultrasonic waves, using a 6 mm titanium alloy probe were irradiated in the juice at a maximum power of 455 W and frequency of 20 kHz. The subsequent effects on biochemical attributes were studied using response surface methodology. Inactivation rates of 90.72 % and 73.18 %, respectively, for PPO and POD enzymes, were observed at the highest US intensity and exposure time. Numerical optimisation using the three-factor, three-level Box-Behnken design suggested that an optimum process at 70 % (energy density: 1610 Wcm⁻²) pulsed at 5 s on and 5 s off for 7 min 30 s resulted in PPO and POD inactivation of the order of 76.42 % and 64.57 % respectively. At these experimental conditions, the optimized levels of biochemical attributes i.e., ascorbic acid (738.50 mg/100 mL), total phenols (17.10 mg/mL), DPPH antioxidant activity (58.47 %), tannins (7.11 µg/mL), colour change (ΔE = 9.04) and flavonoids (6.14 mg/mL) were achieved. The overall statistical models were significant for all the responses except for reducing sugars. Furthermore, the approximation equations for individual responses indicated that the goodness of fit was adequate (R² > 0.90). The results suggested that ultrasound is a suitable processing technique for amla juice stabilisation compared to thermal treatments that result in the loss of quality.     

Design consideration and modelling studies of ultrasound and ultraviolet combined approach for shelf-life enhancement of pine apple juice

Although both ultraviolet (UV) radiation and ultrasound (US) treatment have their capabilities in microbial inactivation, applying any one method alone may require a high dose for complete inactivation, which may affect the sensory and nutritional properties of pineapple juice. Hence, this study was intended to analyse and optimise the effect of combined US and UV treatments on microbial inactivation without affecting the selected quality parameters of pineapple juice. US treatment (33 kHz) was done at three different time intervals, viz. 10 min, 20 min and 30 min., after which, juice samples were subjected to UV treatment for 10 min at three UV dosage levels, viz. 1 J/cm², 1.3 J/cm², and 1.6 J/cm². The samples were evaluated for total colour difference, pH, total soluble solids (TSS), titrable acidity (TA), and ascorbic acid content; total bacterial count and total yeast count; and the standardization of process parameters was done using Response Surface Methodology and Artificial Neural Network. The results showed that the individual, as well as combined treatments, did not significantly impact the physicochemical properties while retaining the quality characteristics. It was observed that combined treatment resulted in 5 log cycle reduction in bacterial and yeast populations while the individual treatment failed. From the optimization studies, it was found that combined US and UV treatments with 22.95 min and1.577 J/cm² ensured a microbiologically safe product while retaining organoleptic quality close to that of fresh juice.     

Ultrasound enhanced production of mycelia and exopolysaccharide by Agaricus bitorquis (Quél.) Sacc. Chaidam

Agaricus bitorquis (Quél.) Sacc. Chaidam (ABSC), is a kind of rare edible macrofungi with a variety of biological ingredients, especially its polysaccharides. However, the low yield limits the popularity and promotion of rare edible macrofungi as well as its macrofungi polysaccharides. Hence, developing a positive and effective cultivation method is of great importance. Herein, an efficient ultrasonic (US) stimulation strategy was developed to improve mycelial growth and exopolysaccharides (EPS) biosynthesis from ABSC in submerged cultivation without light. A time design was employed to illustrate the effect of various process parameters including duration, starting point and times of US irradiation on ABSC productivity. 5 min US treatment for once upon ABSC after fermentation for 48 h could significantly improve EPS production and mycelia growth by above 26% and 15.03%, respectively. Furthermore, six times of 5 min US treatment could make the amount of EPS reach 218.78 ± 17.09 mg/g, which was 2.52-fold higher than that of the control. Moreover, the enhanced effect induced by US was further expounded by fermentation kinetics. Besides, the US treatment could increase mycelia permeability, change structure and reduce mycelial diameter to promote mass transfer, resulting in the improvement of EPS production and mycelia accumulation. The results demonstrated that the present proposed US intensification approach could be useful to boost up the fermentation of ABSC, which possibly applied to yield increase and fermentation product acquisition of macrofungi.     

Combined use of ultrasound-assisted washing with in-package atmospheric cold plasma processing as a novel non-thermal hurdle technology for ready-to-eat blueberry disinfection

Ultrasound (US) has limited disinfection efficacy, and it has been recommended to combine it with chemical disinfectants during fresh produce washing. After washing and before packaging, the disinfection effect of US-assisted washing can be weakened; thus, in-package disinfection is important. As a nutritious fruit, there are no packaged blueberries can be directly eaten. Therefore, in this study, blueberry was selected as the model, and the two most commonly used disinfectants (free chlorine [FC] at 10 ppm and peracetic acid [PAA] at 80 ppm) were combined with low-frequency US (25 kHz) during washing, followed by in-package disinfection using dielectric barrier discharge cold plasma (CP). The disinfection efficacy of US-FC and US-PAA against Escherichia coli O157:H7 and Salmonella Typhimurium was significantly higher than that of US, PAA, or FC alone. The highest disinfection efficacy of CP was observed at the pulse frequency range of 400–800 Hz. For US-FC (1 min) + CP (1 min), an additional 0.86, 0.71, 0.42, and 0.29 log CFU/g of reduction for E. coli O157:H7, S. Typhimurium, aerobic mesophilic counts, and mold and yeast was achieved, respectively, compared with US-FC (2 min) alone. For US-PAA (1 min) + CP (1 min) an additional 0.71, 0.59, 0.32, and 0.21 log CFU/g of reduction was achieved for the above organisms, respectively, compared with US-PAA (2 min) alone. Quality loss (in total color difference, firmness, and anthocyanin content) was not observed after treatment with US-FC + CP, US-PAA + CP, US-FC, or US-PAA. After treatment with US-FC + CP or US-PAA + CP, the reactive oxygen species (ROS) content was significantly lower than that in the other groups, and antioxidant enzyme activity was significantly higher than that in the other groups, suggesting that in-package CP can activate the blueberry antioxidant system to scavenge ROS, thereby lowering the risk of quality loss. US-CP combination not only improves the disinfection efficacy but also lowers quality loss caused by ROS, without prolonging the processing time.     

Combining high intensity ultrasound and experimental design to improve carotenoid extraction efficiency from Buriti (Mauritia flexuosa)

Buriti (Mauritia flexuosa L.) is a significant source of carotenoids, but these compounds have been extracted using laborious and low-effective methods. The present work evaluated the high-intensity ultrasound combined with a chemometric approach to developing an optimal extraction method of carotenoids from buriti pulp. The multivariate optimization was carried out through two steps. First, a simplex-lattice mixture design was used to optimize the extractor solution finding higher extraction yield (903 ± 21 µg g⁻¹) with the acetone:ethanol (75/25) mixture. After, sample mass (80 mg) and sonication time (30 min) were optimized applying central composite design (CCD) which provided a 14% improvement in the extraction method yield. So, the total carotenoid content (TCC) with optimal extraction conditions was 1026 ± 13 µg g⁻¹ which is almost twice the yield of methods known in the literature for buriti. The RP-HPLC-DAD analysis revealed that the carotenoids are gently extracted and β-carotene is the major compound in the extracts. To confirm the accuracy, buriti samples spiked with β-carotene standard and the developed method showed recovery >84% and precision <6.5%. Furthermore, the optimized ultrasound-assisted extraction (UAE) method was applied to other samples (tomato, guava, carrot, mango, acerola, papaya, and pumpkin) and presented a yield to 5.5-fold higher when compared to the reported methods indicating high robustness. Based on results, the UAE method developed has demonstrated feasibility and reliability for the study of carotenoids in buriti pulp as well as in other plant matrices with high biological relevance.     

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.     

Sonication enhances polyphenolic compounds,sugars, carotenoids and mineral elements of apple juice

A study was initiated with the objective of evaluating the effects of sonication treatment on quality characteristics of apple juice such as polyphenolic compounds (chlorogenic acid, caffeic acid, catechin, epicatechin and phloridzin), sugars (fructose, glucose and sucrose), mineral elements (Na, K, Ca, P, Mg, Cu and Zn), total carotenoids, total anthocyanins, viscosity and electrical conductivity. The fresh apple juice samples were sonicated for 0, 30 and 60 min at 20 °C (frequency 25 kHz and amplitude 70%), respectively. As results, the contents of polyphenolic compounds and sugars significantly increased (P < 0.05) but the increases were more pronounced in juice samples sonicated for 30 min whereas, total carotenoids, mineral elements (Na, K and Ca) and viscosity significantly increased (P < 0.05) in samples treated for 60 min sonication. Losses of some mineral elements (P, Mg and Cu) also occurred. Total anthocyanins, Zn and electrical conductivity did not undergo any change in the sonicated samples. Findings of the present study suggest that sonication technique may be applied to improve phytonutrients present naturally in apple juice.     

Contribution of ultrasound and slightly acid electrolytic water combination on inactivating Rhizopus stolonifer in sweet potato

Effects of ultrasound (US, 300, 400, and 500 W) and slightly acidic electrolyzed water (SAEW, 10, 30, and 50 mg/L) combination on inactivating Rhizopus stolonifer in sweet potato tuberous roots (TRs) were investigated. US at 300, 400, and 500 W simultaneous SAEW with available chlorine concentration of 50 mg/L at 40 and 55 °C for 10 min significantly inhibited colony diameters (from 90.00 to 6.00–71.62 mm) and spores germination (p < 0.05). US + SAEW treatment could destroy cell membrane integrity and lead to the leakage of nucleic acids and proteins (p < 0.05). Scanning and transmission electron microscopy results showed that US + SAEW treatment could damage ultrastructure of R. stolonifer, resulted in severe cell-wall pitting, completely disrupted into debris, apparent separation of plasma wall, massive vacuoles space, and indistinct intracellular organelles. US500 + SAEW50 treatment at 40 and 55 °C increased cell membrane permeability, and decreased mitochondrial membrane potential of R. stolonifer. In addition, US500 + SAEW50 at 40 °C and US300 + SAEW50 at 55 °C controlled R. stolonifer growth in sweet potato TRs during 20 days of storage, suggesting effective inhibition on the infection of R. stolonifer. Therefore, US + SAEW treatment could be a new efficient alternative method for storing and preserving sweet potato TRs.     

The disinfection effect of a novel continuous-flow water sterilizing system coupling dual-frequency ultrasound with sodium hypochlorite in pilot scale

In this paper, a self-designed novel continuous-flow water disinfection system coupling dual-frequency ultrasound (US) with chemical disinfectant sodium hypochlorite (NaClO) was tested in a pilot scale using a simulated effluent containing Bacillus subtilis (B. subtilis), one of the indicators of water treatment efficiency. A suspension having a B. subtilis concentration of approximately 10⁴ CFU/mL was introduced into the system to (1) investigate disinfection efficiency of US pretreatment with NaClO (US + NaClO) and simultaneous US and NaClO (US/NaClO) disinfection under different single frequencies; (2) further examine the disinfection efficiency of these two processes with dual-frequency US; and (3) identify dosage reduction of chlorine in this disinfection system. The results demonstrated that lower dual-frequency (17 kHz + 33 kHz) US pretreatment with NaClO disinfection and simultaneous higher dual-frequency (70 kHz + 100 kHz) US and NaClO were beneficial to bacterial inactivation in terms of sterilizing efficiency. It has also been observed that US pretreatment with lower combination of 17 + 33 kHz frequencies showed better enhancement in which log reduction reached to 3.82 after 10 min chlorine reaction (chlorine alone was 0.22 log reduction), nearly 1 log reduction higher than single frequencies at the same constant power. Consequently, at equivalent power dissipation levels, US of lower frequencies combination pretreatment with NaClO disinfection performed such a promising process that one-thirds (from 12 mg/L NaClO reduced to 8 mg/L NaClO) of the required NaClO dosage was reduced for the ideal disinfection efficiency of 4 log reduction, namely 100% disinfection. And the utilization efficiency of NaClO was increased from 37.67% to 85.25% in 30 min of treatment time using an optimized combination of pretreatment and chlorination.     

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.     

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.     

Ultrasound assisted modulation of yeast growth and inactivation kinetics

The yeast Saccharomyces cerevisiae is well known for its application in the food industry for the purpose of developing fermented food. The ultrasound (US) technology offer a wide range of applications for the food industry, including the enhancement of fermentation rates and inactivation of microbial cells. However, a better understanding and standardization of this technology is still required to ensure the scaling-up process. This study investigated the effect of the US technology on the growth of S. cerevisiae using frequencies of 20, 25, 45 and 130 kHz, treatment periods from 2 to 30 min. Furthermore, yeast kinetics subjected to US treatments were evaluated using modelling tools and scanning electron microscopy (SEM) analysis to explore the impact of sonication on yeast cells. Yeast growth was monitored after different US treatments plotting optical density (OD) at 660 nm for 24 h at 30 ⁰C. Growth curves were fitted using models of modified Gompertz and Scale-Free which showed good parameters of the fit. In particular, US frequencies of 45 and 130 kHz did not have a disruptive effect in lag phase and growth rate of the yeast populations, unlike the frequency of 20 kHz. Moreover, inactivation curves of yeast cells obtained after exposure to 20 and 25 kHz also observed the best fit using the Weibull model. US frequency of 20 kHz achieved significant reductions of 1.3 log cfu/mL in yeast concentration and also induced important cell damage on the external structures of S. cerevisiae. In conclusion, the present study demonstrated the significant effect of applying different US frequencies on the yeast growth for potential application in the food industry.     

Effects of thermal treatment and sonication on quality attributes of Chokanan mango (Mangifera indica L.) juice

Ultrasonic treatment is an emerging food processing technology that has growing interest among health-conscious consumers. Freshly squeezed Chokanan mango juice was thermally treated (at 90 °C for 30 and 60 s) and sonicated (for 15, 30 and 60 min at 25 °C, 40 kHz frequency, 130 W) to compare the effect on microbial inactivation, physicochemical properties, antioxidant activities and other quality parameters. After sonication and thermal treatment, no significant changes occurred in pH, total soluble solids and titratable acidity. Sonication for 15 and 30 min showed significant improvement in selected quality parameters except color and ascorbic acid content, when compared to freshly squeezed juice (control). A significant increase in extractability of carotenoids (4–9%) and polyphenols (30–35%) was observed for juice subjected to ultrasonic treatment for 15 and 30 min, when compared to the control. In addition, enhancement of radical scavenging activity and reducing power was observed in all sonicated juice samples regardless of treatment time. Thermal and ultrasonic treatment exhibited significant reduction in microbial count of the juice. The results obtained support the use of sonication to improve the quality of Chokanan mango juice along with safety standard as an alternative to thermal treatment.     

Impact of different ultrasound-assisted processes for preparation of collagen hydrolysates from Asian bullfrog skin on characteristics and antioxidative properties

This study focused on impact of ultrasound-assisted process (UAP) at pre-treatment (UP) and simultaneous treatment (US) during papain hydrolysis for preparing collagen hydrolysate (CH) from Asian bullfrog skin. Ultrasonication times were varied (10, 20, 30 min), and CH prepared using papain hydrolysis without UAP was used as control. Different UAPs provided CH with various hydroxyproline contents, α-amino group contents, surface hydrophobicities, and antioxidative activities. UP at 20 min (UP-20) and US at 30 min (US-30) provided highly antioxidative CHs, which were selected for further studies on their Oxygen reactive absorbance capacity (ORAC) and molecular characteristics. CHs from UP-20 and US-30 had higher ORAC than that of control group (p ≤ 0.05). Slight difference in amino acid composition was detected between samples. Based on these results, molecular characteristic styles, molecular weight profile, antioxidative peptide content, and secondary structure of each sample were obtained. These results indicate that UP and US used varied enzymatic hydrolysis patterns and modified molecular conformation of CH, resulting in enhanced antioxidative activity. Therefore, different UAPs as UP and US could be effectively used in preparation of CH using papain hydrolysis from Asian bullfrog skin, which could improve production process efficiency by enhancing their bioactivity, particularly antioxidative activity.     

Comparison of ultrasound-assisted extraction with conventional extraction methods of oil and polyphenols from grape (Vitis vinifera L.) seeds

Ultrasound-assisted extraction (US) carried out at 20 KHz, 150 W for 30 min gave grape seed oil yield (14% w/w) similar to Soxhlet extraction (S) for 6 h. No significant differences for the major fatty acids was observed in oils extracted by S and US at 150 W. Instead, K₂₃₂ and K₂₆₈ of US- oils resulted lower than S-oil. From grape seeds differently defatted (S and US), polyphenols and their fractions were extracted by maceration for 12 h and by ultrasound-assisted extraction for 15 min. Sonication time was optimized after kinetics study on polyphenols extraction. Grape seed extracts obtained from seeds defatted by ultrasound (US) and then extracted by maceration resulted the highest in polyphenol concentration (105.20 mg GAE/g flour) and antioxidant activity (109 Eq αToc/g flour).     

Effect of ultrasound and high hydrostatic pressure (US/HHP) on the degradation of dextran catalyzed by dextranase

In our current research work, the effect of combination of ultrasonic irradiation and high hydrostatic pressure (US/HHP) on the enzymatic activity and enzymatic hydrolysis kinetic parameters of dextran catalytic by dextranase were investigated. Furthermore, the effects of US/HHP on the structure of dextranase were also discussed with the aid of fluorescence spectroscopy and circular dichroism (CD) spectroscopy. The maximum hydrolysis of dextran was observed under US (40 W at 25 kHz for 15 min) combined with HHP (400 MPa for 25 min), in which the hydrolysis of dextran increased by 163.79% compared with the routine thermal incubation at 50 °C. Results also showed that, V_max and K_M values, as well as, k_cat of dextranase under US/HHP treatment were higher than that under US, HHP and thermal incubation at 50 °C, indicated that, the substrate is converted into the product at an increased rate when compared with the incubation at 50 °C. Compared to the enzymatic reaction under US, HHP, and routine thermal incubation, dextranase enzymatic reaction under US/HHP treatment showed decreases in Ea, ΔG and ΔH, however small increase in ΔS value was observed. In addition, fluorescence and CD spectra reflected that US/HHP treatment had increased the number of tryptophan on dextranase surface with increased α-helix by 19.80% and reduced random coil by 6.94% upon US/HHP-treated dextranase protein compared to the control, which were helpful for the improvement of its activity. These results indicated that, the combination of US and HHP treatments could be an effective method for improving the hydrolysis of dextran in many industrial applications including sugar manufacturing processes.     

Effects of phonophoresis with Arnica montana onto acute inflammatory process in rat skeletal muscles: An experimental study

This study aimed at verifying the effects of phonophoresis associated with Arnica montana on the acute phase of an inflammatory muscle lesion. Forty Wistar male rats (300 ± 50 g), of which the Tibialis Anterior muscle was surgically lesioned, were divided into four groups (n = 10 each): control group received no treatment; the ultrasound group (US) was treated in pulsed mode with 1-MHz frequency, 0.5 W/cm² intensity (spatial and temporal average – SATA), duty cycle of 1:2 (2 ms on, 4 ms off, 50%), time of application 3 min per session, one session per day, for 3 days; the phonophoresis or ultrasound plus arnica (US+A) group was treated with arnica with the same US parameters plus arnica gel; and the arnica group (A) was submitted to massage with arnica gel, also for 3 min, once a day, for 3 days. Treatment started 24 h after the surgical lesion. On the 4th day after lesion creation, animals were sacrificed and sections of the lesioned, inflamed muscle were removed for quantitative (mononuclear and polymorphonuclear cell count) and qualitative histological analysis. Collected data from the 4 groups were statistically analyzed and the significance level set at p < 0.05. Results show higher mononuclear cell density in all three treated groups with no significant difference between them, but values were significantly different (p < 0.0001) when compared to control group’s. As to polymorphonuclear cell density, significant differences were found between control group (p = 0.0134) and US, US+A and A groups; the arnica group presented lesser density of polymorphonuclear cells when compared (p = 0.0134) to the other groups. No significant difference was found between US and US+A groups. While the massage with arnica gel proved to be an effective anti-inflammatory on acute muscle lesion in topic use, these results point to ineffectiveness of Arnica montana phonophoresis, US having seemingly checked or minimized its anti-inflammatory effect.