Pulsed multifrequency thermosonication induced sonoporation in Alicyclobacillus acidoterrestris spores and vegetative cells.

Alicyclobacillus acidoterrestris (AAT) was proposed as an index of pasteurization design for high-acid fruit products due to its spore resistance and repeated spoilage incidences in fruit juices. This study aimed to determine the effectiveness of pulsed multifrequency ultrasound to minimize AAT spores and vegetative cells in aqueous suspension. For this research, an investigation of the reactive oxygen species and antioxidant activity was performed to examine the effect of temperature and frequency on AAT spore inoculation. Total decreases in AAT bacteria were 5.99, 5.74 Log CFU/mL in vegetative cells for dual-frequency thermosonication (DFTS) and dual-frequency ultrasonication (DFUS), respectively, while 5.90 and 5.38 Log CFU/mL in spores for both DFUS and DFTS, respectively. The loss of the percentage of cells in ultrasound (US) and thermosonication (TS) treatments was inversely associated with the rate of O2-and H2O2 development. The fluorescence microscopy revealed a higher bactericidal efficacy of DFTS compared to the DFUS and control. Scanning Electron Microscopy (SEM) and Transmission electron microscopy (TEM) demonstrated ultra-structural modifications such as the interruption of cell walls by cavitation and pores in the membrane structure of the AAT bacteria induced by sonoporation. Several TS frequencies of 20/40/60, 20/40, and 20 kHz treated spores had a higher electrical conductivity than untreated ones, with an improvement of 7.94, 5.68, and 3.72 %, respectively. Fourier-transform infrared (FTIR) spectroscopy revealed major changes in the spectral region of membrane fatty acids and proteins of AAT. Simultaneously, AAT inactivation specific energy rate was significantly reduced using dual-frequency ultrasound, compared to mono-frequency thermosonication. The significant results of this work recommended pulsed DFUS as an alternative application to mono-frequency US in beverage industries.

Quality attributes optimization of orange juice subjected to multi-frequency thermosonication: Alicyclobacillus acidoterrestris spore inactivation and applied spectroscopy ROS characterization.

This is the first time to investigate the synergistic inactivation effect and mechanism of multifrequency ultrasound (MTUS) on A. acidoterrestris (AAT) vegetative cells and spores, nutrients and enzymes of orange juice. The optimized results of MTUS (using Box Behnken design- surface responsemethodology) and further comparison with different mode of ultrasound (mono-and multi-frequency) revealed that 20/40 kHz, 24 min and 64 °C were the best optimum results. The AAT spores and vegetative cells were inactivated by 2 and 4 logs, respectively, without deteriorating orange juice contents. In addition, AAT inactivation indicated an inversely proportional relationship with ROS production. FT-IR and UV-Vis spectroscopy characterization confirmed the existence of ROS in treated orange juice and LF-NMR analysis confirmed the inactivation of AAT spores. The findings illustrated the successfully used dual-frequency ultrasound technology for fruit beverages, promoting beneficial changes in physical properties without any significant effects on the quality of ascorbic acid.

Kinetic modeling of inactivation of natural microbiota and Escherichia coli on cherry tomato treated with fixed multi-frequency sonication.

The suitability of some non-linear kinetic models (Weibull {with or without tail}, Log-linear, Log-linear shoulder {with or without tail}, Biphasic linear, Logistic, Multi-target and Single-target models) were evaluated to determine the inactivation kinetics of inoculated E. coli, and natural microbiota (i.e. mesophilic aerobic bacteria, and mold and yeast) on cherry tomato treated with fixed multi-frequency ultrasound. Almost all the studied model fitted well (R2 ≥ 0.9) for the inactivation kinetics; however, the Weibull, Log-linear shoulder, and Biphasic linear model showed the highest statistical parameters (0.9 ≤ adj. R2 ≤ 0.99 and smallest RMSE and SSE values). All the three models could be used to compare the kinetic behavior of E. coli and the first two models for the kinetic behavior of mesophilic aerobic bacteria and mold and yeast during sonication treatment. Two distinctive inactivation curves were obtained for the mono-frequency and the multi-frequency (dual and tri-frequency) for all the microbial inactivation. The remarkable results obtained for dual and tri-frequency sonication shows to be an effective and promising alternative to the traditional microbial inactivation techniques and the common practice of using ultrasound with other sanitizing methods.

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.

Storage effects on the quality quartet of orange juice submitted to moderate thermosonication: Predictive modeling and odor fingerprinting approach.

The effects of moderate thermosonication (MTS) on the quality quartet: physico-chemical, microbial, nutritional and sensory qualities of orange juice (OJ) inoculated with Alicyclobacillus acidoterrestris (AAT) were studied during 24 days of storage at ambient and refrigerated temperatures. The bioactive compounds and antioxidant activity of OJ decreased with storage, while the pectin methyl esterase (PME) increased. Nonetheless, noticeable changes were observed from the 12th day of storage. There was no obvious (p > 0.05) variation in pH and total soluble solids. To determine the nutritional and microbial quality characteristics of OJ during storage, non-linear kinetic curves were successfully fitted with least square fitting polynomial and four-parameter log-logistic distribution models. The E-nose sensors succeeded in discriminating between the aroma of non-treated and treated OJ based on linear discriminant analysis (LDA). Furthermore, terpenes, alcohol and partially aromatic compounds were the main spoilage indicators of OJ during storage based on E-nose analysis and confirmed by HS-SPME-GC/MS analysis. Thus, MTS significantly extended the shelf life of the quality quartet of natural OJ at 4 °C. E-nose-GC/MS fusion offered odor fingerprints to AAT microorganisms that can be used as spoilage index without using traditional food analysis techniques. The proposed approach can be used as an alternative tool for rapid detection of spoilage microorganisms in OJ.

Acoustically-aided osmo-dehydration pretreatments under pulsed vacuum dryer for apple slices: drying kinetics, thermodynamics, and quality attributes.

The research work investigates the effect of different pretreatment conditions (osmotic dehydration [OD], ultrasound [US], and ultrasound-assisted osmotic dehydration [UOD]) on the drying kinetics modeling, thermodynamics, weight reduction, degradation kinetics of vitamin C, and color of apple slices under pulsed vacuum dryer (PVD). The findings showed that UOD pretreatment significantly improved drying time and increased weight reduction comparative to OD, US, and the control sample. Drying kinetics modeling revealed that the Hii model better described the drying kinetic behavior of the apple slices than the other nine mathematical models based on higher coefficient of determination (R2 ), root mean square error (RMSE), and reduced chi-square (χ2 ). Analysis of vitamin C content revealed a 46.05%, 31.28%, and 25.95% retention for UOD, US, and OD, respectively, after drying. Second-order kinetics could accurately predict the degradation kinetics of vitamin C compared to first-order kinetics. Vitamin C degradation kinetics showed lower k-value, higher D-value (time required for 90% degradation), and half-life indicating a higher retention of vitamin C content for UOD pretreatment compared to OD and US during drying. L*, b*, and chroma values of UOD were significantly (P < 0.05) higher compared to US, OD, and the control sample. PRACTICAL APPLICATION: The findings of this study revealed that ultrasonic-aided osmotic dehydration is a unique and novel pretreatment technique prior to drying, which significantly shortens drying time as a result of faster moisture/mass transfer, improves processing efficiency thereby reducing processing cost, improves quality parameters, and preserve phytochemicals. This makes the business operations of food processors competitive and as well provide value for customers.