Comparative evaluation of the effects of pulsed electric field and freezing on cell membrane permeabilisation and mass transfer during dehydration of red bell peppers

The extent of cell membrane permeabilisation due to high intensity electric field pulses (HELP) varying number of pulses (1–50) using electric field of 2 kV/cm, 400 μs pulse duration and freezing on mass transfer and vitamin C content during osmotic (50° Brix sucrose at 40 °C) and convective air (60 °C, 1 m/s for 5 h) dehydration of red bell peppers was studied. Total pore area due to HELP increased with number of pulses while freezing resulted in total pore area of almost 6 times as greater as the highest value from the HELP process. Higher water loss was observed for all HELP treated than for prefrozen samples while slow freezing provided samples with the highest solids uptake. The correlation coefficient (R²) of linear regression between water loss and solids gain estimated from either total solids or soluble solids measurement ranged from 0.954 to 0.998 suggesting the possibility of using the soluble solids method in evaluating mass transfer kinetics during osmotic dehydration process. Drying rate during convective air-drying was more enhanced by HELP than by freezing. Electrical conductivity of the osmotic solution increased with the degree of permeabilisation to a given medium value after which no further increase in the release of the intracellular ions was observed. Minimal vitamin C depletion was observed immediately after HELP treatment. The order of magnitude of vitamin C retention was untreated>frozen>HELP pretreated samples with 1 pulse>5 pulses>50 pulses>10 pulses>20 pulses after osmotic dehydration. The reduction in vitamin C content of HELP treated samples after convective drying ranged from approximately 11 to 24% while freezing resulted in approximately 24% decrease compared to the untreated samples.     

Effects of high hydrostatic pressure or high intensity electrical field pulse pre-treatment on dehydration characteristics of red paprika

The effects of various pre-treatments (hot water blanching, skin treatments, high pressure and high intensity electric field pulse treatment) on the dehydration characteristics of red paprika (Capsicum annuum L.) were evaluated and compared with untreated samples. Hot water blanching (100°C, 3 min) prior to dehydration (fluidised bed dryer at 60°C, 6 h and 1 m/s) resulted in the permeabilisation of 88% of the cell membranes in paprika, which in turn resulted in a higher mass and heat transfer. Skin treatments (such as lye peeling and acid treatment), as practised conventionally, increased dehydration rates but affected only the skin permeability. The application of high hydrostatic pressure (HHP, 400 MPa for 10 min at 25°C) or high intensity electric field pulses (HELP, 2.4 kV/cm, pulse width 300 μs, 10 pulses, pulse frequency 1 Hz) pre-treatments resulted in cell disintegration indexes of 0.58 and 0.61, respectively. Cell permeabilisation of these physical treatments resulted in higher drying rates, as well as higher mass and heat transfer coefficients, as compared to conventional pre-treatments.     

Anwendung elektrischer Hochspannungsimpulse als Vorbehandlungsverfahren zur Beeinflussung der Trocknungscharakteristika und Rehydratationseigenschaften von Kartoffelwürfeln

High intensity electric field pulses as pretreatment for affecting dehydration characteristics and rehydration properties of potato cubes. High intensity electric field pulse (HELP) treatment was evaluated as pretreatment to possibly affect dehydration and rehydration characteristics of potato cubes (1 × 1 × 1 cm). Effects of electric field strength E = 0.35-3.0 kV · cm^?1 and number of pulses n = 1-70 on the degree of permeabilization of potato cells was evaluated using a centrifugal method. Optimization of E and n regarding maximum permeabilization of cells was performed. Energy requirements were found to be low (6.4-16.2 kJ · kg^?1) resulting in an increase in temperature in the treated product of _ΔT = 1.8-4.5°C. The HELP treatment enabled a maximum degree of permeabilization and resulted in a cell liquid release from potato cubes after centrifugation (700 × g) to up to 27-29% (untreated control 0%) as related to total water content. Treatment with optimum parameters (E = 0.9-2.0 kV · cm^?1, n = 15-30) resulted in improved mass transfer within the product and drying time during fluidized bed drying (T_tr = 55°C, 70°C, air velocity 2 m · s^?1) of potato cubes could be reduced to up to 20-25%. Rehydration properties and quality characteristics of the products after cooking were not affected by the high intensity electric field pulse pretreatment.     

Milk thermization by pulsed electric fields (PEF) and electrically induced heat

Whole milk was processed using selected combinations of pulsed electric fields (PEF) and thermal treatments to inactivate Listeria innocua. Electric field intensities of 30 and 40 kV/cm were applied at selected number of pulses (1–30) and temperatures (20–72 °C) for less than 10 s. A maximum microbial reduction of 4.3 log cycles was achieved using 10, 17.5, 20 and 25 pulses, when processing milk at 30 kV/cm and initial temperatures of 43, 33, 23 and 13 °C, respectively. Around 4.3 log cycles of L. innocua was observed when treating milk at 40 kV/cm using 3, 10, 12.5, 15, and 20 pulses and 53, 33, 23, 15, and 3 °C, respectively. Milk treated with 40 kV/cm of electric field intensity, few pulses, and initial temperature close to 55 °C showed the best balance between L. innocua inactivation and energy-consumption. An energy expenditure of around 244 J/mL was achieved, which can be further reduced to 44 J/mL using a thermal regeneration system.     

Reduction of Protease Activity in Simulated Milk Ultrafiltrate by Continuous Flow High Intensity Pulsed Electric Field Treatments

A protease from Bacillus subtilis suspended in simulated milk ultrafiltrate (SMUF) was subjected to high intensity pulsed electric field (HIPEF) treatments of up to 6787 kJ/l, applying field strengths ranging from 19.7 to 35.5kV/cm up to 896 μs to evaluate the feasibility of this treatment on inactivating the enzyme. In addition, the influence of the pulse repetition rate (67, 89, and 111 Hz) and pulse width (4 and 7 μs) on the effectiveness of HIPEF treatments was tested. Protease activity was considerably reduced; a maximum inactivation of 62.7% was achieved after an 896‐μS treatment at 35.5 kV/cm and 111 Hz. Protease activity decreased exponentially with increase of input energy density, treatment time, field strength, and pulse repetition rate when exposed to HIPEF processing.     

Inactivation of microorganisms using pulsed electric fields: the influence of process parameters on Escherichia coli,Listeria innocua,Leuconostoc mesenteroides and Saccharomyces cerevisiae

This study examines the killing effect of pulsed electric fields (PEF) on four organisms suspended in a model medium. Escherichia coli, Listeria innocua, Leuconostoc mesenteroides and Saccharomyces cerevisiae differ in size, shape and cell wall construction. The electric field strength, pulse duration and number of pulses were varied in the ranges of 25–35 kV/cm, 2–4 μs and 20–40 pulses, respectively. The results showed that S. cerevisiae was the most sensitive organism with a 6-log reduction, followed by E. coli with a 5.4-log reduction, when they were exposed to 30 kV/cm, and 20 pulses with 4 μs duration. The most resistant organisms were L. innocua and L. mesenteroides with only a 3-log reduction, however, by increasing the parameters to 35 kV/cm and 40 pulses with 4 μs pulse duration; marked viability reductions of 8 and 7 log, respectively, were observed. Heat, which is generated during the process, has limited killing effect on the cells, hence the observed reduction can be ascribed to the PEF treatment. Although transmission electron microscopy of PEF treated cells did not confirm membrane damage, observations suggest that PEF treatments have profound direct or indirect effects on the intracellular organisation of microorganisms.     

Comparative study on shelf life of whole milk processed by high-intensity pulsed electric field or heat treatment

The effect of high-intensity pulsed electric fields (HI-PEF) processing (35.5 kV/cm for 1,000 or 300 μ with bipolar 7-μs pulses at 111 Hz; the temperature outside the chamber was always < 40° C) on microbial shelf life and quality-related parameters of whole milk were investigated and compared with traditional heat pasteurization (75° C for 15 s), and to raw milk during storage at 4° C. A HIPEF treatment of 1,000 μ ensured the microbiological stability of whole milk stored for 5 d under refrigeration. Initial acidity values, pH, and free fatty acid content were not affected by the treatments; and no proteolysis and lipolysis were observed during 1 wk of storage in milk treated by HIPEF for 1,000 μ. The whey proteins (serum albumin, β-lactoglobulin, and α-lactalbumin) in HIPEF-treated milk were retained at 75.5, 79.9, and 60%, respectively, similar to values for milk treated by traditional heat pasteurization.     

Plasmin Inactivation with Pulsed Electric Fields

Plasmin (fibrinolysin E.C.3.4.21.7), an indigenous enzyme in bovine milk, added to simulated milk ultrafiltrate (SMUF) at 100 μg/mL (pH 6.11 and ionic strength 0.056 M) was treated at 10°C and 15°C with pulsed electric fields (HVPEF) of 15, 30 and 45 kV/cm and number of pulses 10, 20, 30, 40 and 50. The plasmin activity measured using a commercial assay, was reduced 90% after 50 pulses at both 30 and 45 kV/cm and at a processing temperature of 15°C. Similar inactivation was obtained when plasmin (100 μg/mL) in SMUF was heated at 40°C for 15 min. Inactivation of the enzyme depended on the number of pulses applied during treatment, intensity of the applied field, and processing temperature.     

In vitro bioaccessibility of isoflavones from a soymilk-based beverage as affected by thermal and non-thermal processing

High Pressure Processing (HPP) and High Intensity Pulsed Electric Fields (HIPEF) are non-thermal processing technologies used for obtaining safe and high-quality foods and beverages. In the present work, the changes on both the concentration and the bioaccessibility of isoflavones from treated (thermally and non-thermally) and untreated soymilk-based beverages were evaluated. Thermal treatment (TT) was applied at 90 °C for 1 min, HPP: 400 MPa at 40 °C for 5 min and HIPEF: 35 kV cm⁻¹ with 4 μs bipolar pulses at 200 Hz for 1800 μs. Later, beverages were subjected to an in vitro gastrointestinal digestion for obtaining the bioaccessibility. Thermal and non-thermal processing increased the isoflavone concentration up to 25–26% in TT and HIPEF treated beverages, and up to 38.52% in HPP treated. After in vitro digestion, the concentration of isoflavones in non-thermally processed beverages was higher (70.55% for HIPEF and 98.77% for HPP) than that TT processed (18.52%). HIPEF processing and HPP increased the isoflavone bioaccessibility up to 35.40 and 47.32%, respectively, regarding the untreated product. These results demonstrate that both non-thermal processing technologies HIPEF and HPP are suitable for obtaining high quality and nutritious beverages by improving their isoflavone bioaccessibility.     

INACTIVATION OF ESCHERICHIA COLI IN SKIM MILK BY HIGH INTENSITY PULSED ELECTRIC FIELDS

The inactivation of microorganisms is the most important function in the processing of milk and dairy products. Traditionally, this purpose is realized by thermal treatment, but heat produces alterations to flavor and taste in addition to nutrient loss. The high intensity pulsed electric field (PEF) treatment should be a good alternative to heat because demonstrations have shown PEF can reduce the Escherichia coli survival fraction in aqueous solutions and model foods. In this study, PEF treatment was found to inactivate E. coli in skim milk (inoculum 10⁹ CFU/mL) at 15C. The microorganism inactivation satisfied Hülsheger's model following a first order kinetic for both the electric field intensity and number of pulses when skim milk inoculated with E. coli was treated in a static or continuous flow chamber. PEF treatment in a continuous system when the critical electric field (E_c) and minimum number of pulses (n_min) were 12.34 kV/cm and 2.7 at 30 kV/cm and 30 pulses (0.7–1.8 μs pulse width) inactivated more microorganisms than in a static system. It has also been proven that increasing the pulse duration increases the E. coli inactivation. The inactivation of E. coli using PEF is more limited in skim milk than in a buffer solution when exposed to similar treatment conditions of field intensity and number of pulses due to the complex composition of skim milk, its lower electrical resistivity and the presence of proteins.     

Predicting inactivation of Salmonella senftenberg by pulsed electric fields

Pulsed electric field inactivation of Salmonella senftenberg suspended in McIlvaine buffer of pH 7 and conductivity 2 mS/cm was investigated. In this study, square wave waveform pulses were used. After the same treatment time, inactivation of S. senftenberg depended neither on pulse width (1–15 μs) nor frequency of treatment (1–5 Hz). Survivor curves of S. senftenberg at different electric field strengths did not follow first-order kinetics. These survival curves were described by the log-logistic model proposed by Cole et al. [Cole, M. B., Davies, K. W., Munro, G., Holyoak, C. D., and Kilsby, D. C. (1993). A vitalistic model to describe the thermal inactivation of Listeria monocytogenes. Journal of Industrial Microbiology, 12, 232–239]. Comparison of measured and estimated values showed that this model accurately described the inactivation of S. senftenberg by high electric field pulses in the range of 12–28 kV/cm.     

Equivalent processing for pasteurization of a pineapple juice–coconut milk blend by selected nonthermal technologies

Identifying equivalent processing conditions is critical for the relevant comparison of food quality attributes. This study investigates equivalent processes for at least 5-log reduction of Escherichia coli and Listeria innocua in pineapple juice–coconut milk (PC) blends by high-pressure processing (HPP), pulsed electric fields (PEF), and ultrasound (US) either alone or combined with other preservation factors (pH, nisin, and/or heat). The two blends (pH 4 and 5) and coconut milk (pH 7) as a reference were subjected to HPP at 300–600 MPa, 20°C for 0.5–30 min; PEF at an electric field strength of 10–21 kV/cm, 40°C for 24 µs; and US at 120 µm amplitude, 25 or 45°C for 6 or 10 min. At least a 5-log reduction of E. coli was achieved at pH 4 by HPP at 400 MPa, 20°C for 1 min; PEF at 21 kV/cm, 235 Hz, 40°C for 24 µs; and US at 120 µm, 45°C for 6 min. As L. innocua showed greater resistance, a synergistic lethal effect was provided at pH 4 by HPP with 75 ppm nisin at 600 MPa, 20°C for 5 min; PEF with 50 ppm nisin at 18 kV/cm, 588 Hz, 40°C for 24 µs; and US at 45°C, 120 µm for 10 min. The total soluble solids (11.2–12.4°Bx), acidity (0.47%–0.51% citric acid), pH (3.91–4.16), and viscosity (3.55 × 10⁻³–4.0 × 10⁻³ Pa s) were not significantly affected under the identified equivalent conditions. HPP was superior to PEF and US, achieving higher ascorbic acid retention and lower color difference in PC blend compared to the untreated sample.     

Inactivation of Saccharomyces cerevisiae and Bacillus cereus by pulsed electric fields technology

The effect of pulsed electric field (PEF) treatment, applied in a continuous system, on Saccharomyces cerevisiae and Bacillus cereus cells and spores was investigated. S. cerevisiae inoculated into sterilised apple juice and B. cereus cells and spores inoculated into sterilised 0.15% NaCl were treated with electric field strengths of 10–28 kV/cm using an 8.3 pulse number and with pulse numbers of 4.2–10.4 at 20 kV/cm, respectively. The inactivation of S. cerevisiae depended on the electric field intensity and number of pulses. The yeast inactivation increased when the applied electric field intensity and pulse number were increased. Approximately four log cycles reduction was achieved in apple juice using 10.4 pulses at 20 kV/cm. B. cereus cells were less sensitive to PEF treatment. The reduction in microbial count of B. cereus cells was hardly more than one log cycle using 10.4 pulses at 20 kV/cm. The applied PEF treatment was ineffective on Bacillus cereus spores.     

Reduction of protease activity in milk by continuous flow high-intensity pulsed electric field treatments

High-intensity pulsed electric field (HIPEF) is a non-thermal food processing technology that is currently being investigated to inactivate microorganisms and certain enzymes, involving a limited increase of food temperature. Promising results have been obtained on the inactivation of microbial enzymes in milk when suspended in simulated milk ultrafiltrate. The aim of this study was to evaluate the effectiveness of continuous HIPEF equipment on inactivating a protease from Bacillus subtilis inoculated in milk. Samples were subjected to HIPEF treatments of up to 866 micros of squared wave pulses at field strengths from 19.7 to 35.5 kV/cm, using a treatment chamber that consisted of eight colinear chambers connected in series. Moreover, the effects of different parameters such as pulse width (4 and 7 micros), pulse repetition rates (67, 89, and 111 Hz), and milk composition (skim and whole milk) were tested. Protease activity decreased with increased treatment time or field strength and pulse repetition rate. Regarding pulse width, no differences were observed between 4 and 7 micros pulses when total treatment time was considered. On the other hand, it was observed that milk composition affected the results since higher inactivation levels were reached in skim than in whole milk. The maximum inactivation (81%) was attained in skim milk after an 866-micros treatment at 35.5 kV/cm and 111 Hz.     

Nature of the inactivation of Escherichia coli suspended in an orange juice and milk beverage

The killing effect of pulsed electric fields (PEF) on Escherichia coli (ATCC 8739) suspended in an orange juice and milk beverage was studied. Bipolar square pulses with a pulse width of 2.5 μs were applied. Electric field strength and treatment times ranged from 15 to 40 kV/cm, and from 0 to 700 μs, respectively. A maximum of 3.83 log reductions was achieved at 15 kV/cm and 700 μs. The experimental data were fitted to Bigelow and Hülsheger models and Weibull distribution function. Results indicated that Weibull function best described the experimental data (lowest mean square error). As there were no significant differences in the values of the shape factor (n) at the electric field strength of 25–40 kV/cm, the number of parameters in the Weibull model were reduced, leading to a simplified model with a fit similar to that obtained with the full model.     

Inactivation of Pseudomonas fluorescens in skim milk by combinations of pulsed electric fields and organic acids

Pseudomonas fluorescens suspended in skim milk was inactivated by application of pulsed electric fields (PEF) either alone or in combination with acetic or propionic acid. The initial concentration of microorganisms ranged from 10(5) to 10(6) CFU/ml. Addition of acetic acid and propionic acid to skim milk inactivated 0.24 and 0.48 log CFU/ml P. fluorescens, respectively. Sets of 10, 20, and 30 pulses were applied to the skim milk using exponentially decaying pulses with pulse lengths of 2 micros and pulse frequencies of 3 Hz. Treatment temperature was maintained between 16 and 20 degrees C. In the absence of organic acids, PEF treatment of skim milk at field intensities of 31 and 38 kV/cm reduced P. fluorescens populations by 1.0 to 1.8 and by 1.2 to 1.9 log CFU/ml, respectively. Additions of acetic and propionic acid to the skim milk in a pH range of 5.0 to 5.3 and PEF treatment at 31, 33, and 34 kV/cm, and 36, 37, and 38 kV/cm reduced the population of P. fluorescens by 1.4 and 1.8 log CFU/ml, respectively. No synergistic effect resulted from the combination of PEF with acetic or propionic acid.     

Effects of high intensity pulsed electric field and thermal treatments on a lipase from Pseudomonas fluorescens

Milk and dairy products may contain microorganisms capable of secreting lipases that cause sensory defects and technological problems in the dairy industry. In this study, the effects of thermal and high-intensity pulsed electric field (HIPEF) treatments on an extracellular lipase from Pseudomonas fluorescens, suspended in a simulated skim milk ultrafiltrate (SMUF) have been evaluated. Heat treatments applied were up to 30 min from 50 to 90 degrees C. HIPEF treatments were carried out using pilot plant facilities in a batch or continuous flow mode, where treatment chambers consisted of parallel and coaxial configuration, respectively. Samples were subjected to up to 80 pulses at electric field intensities ranging from 16.4 to 37.3 kV/cm. This resulted in a lipase that was quite resistant to heat and also to HIPEF. High (75 degrees C-15 s) and low pasteurization treatments (63 degrees C-30 min) led to inactivations of 5 and 20%, respectively. Using the batch-mode HIPEF equipment, a 62.1% maximum activity depletion was achieved after 80 pulses at 27.4 kV/cm. However, when HIPEF treatments were applied in the continuous flow mode, an inactivation rate of just 13% was achieved, after applying 80 pulses at 37.3 kV/cm and 3.5 Hz. The results of both heat and HIPEF treatments on enzyme inactivation were adjusted with good agreement to a first-order kinetic model (R2 > 62.3%).     

Osmotic dehydration of carrot tissue enhanced by pulsed electric field,salt and centrifugal force

The osmotic dehydration (OD) kinetics of carrot disc untreated and treated by pulsed electric field (PEF) was studied under centrifugation (2400 × g), stirring (250 rpm) and with a salt addition (NaCl/sucrose solutions 0%/65%, 5%/60% and 15%/50%). The PEF intensity was E = 0.60 kV/cm and the treatment duration was t_PEF = 0.05 s (500 rectangular monopolar pulses each of 100 μs). The water loss (WL), solids gain (SG) and water loss/solids gain ratios (WL/SG) were evaluated in the binary (sucrose + water) and ternary (sucrose + salt + water) solutions at the temperature of 20 °C during 4 h. The mass ratio of sample to solution was 1:3. The PEF treatment and salt addition enhanced the OD kinetics. WL and SG were increased under centrifugation (centrifugal OD) and under stirring (static OD). The centrifugal field enhanced the WL, however, decreased the SG comparing to the static OD. Therefore, the static OD has advantages for the higher SG (confectionary adds), while the centrifugal OD is better appropriated if the WL should be increased and the solids (sugar) uptake should be limited (dietetic products).The two-exponential kinetic model fitted well to experimental data for both static and centrifugal OD. The correlation coefficient was R² = 0.982–0.999 and the standard error was 5–10%.     

Indoxylsulfate in milk

Indoxylsulfate in 27 individual milk samples ranged from 25.4 to 111 μg/l (average 52.3 μg/l); pooled milk samples from 12 farms contained 81.1 μg/l (46.4–146 μg/l); the variation in indoxylsulfate concentration of dried skimmed milk over a period of one year amounted to 23%. This variability is likely attributable to regional and seasonal, and hence to feeding effects. The indoxylsulfate content of milk seems also to be dependent upon the degree of fermentation during processing of milk; yoghurt contained very low amounts of this component (6.4 μg/kg). On the other hand, heat treatment of the milk (HTST, UHT, sterilization) apparently does not affect its indoxylsulfate content. Indoxylsulfate concentrations in milk correlated positively with blood-serum indoxylsulfate content (r=0.752,n=20) and with the urea content of milk (r=0.61,n=12 pooled milks). Further research is suggested on the use of indoxylsulfate determinations as an aid to determine sweet whey added to dried skimmed milk, also as an analytical tool to differentiate bovine and sheep milks.     

Impact of the electric field intensity and treatment time on whey protein aggregate formation

Whey proteins are being integrated as high-value food product ingredients due to their versatile and tunable techno-functionality. To meet high food quality and clean label expectations by consumers, electric field (EF) technologies have been proposed to open new frontiers in this field. Despite a variety of studies, it remains ambiguous which EF parameters are crucial to achieving targeted whey protein modifications. Reconstituted liquid whey protein concentrate (WPC_L) and filtered, non-heat-treated liquid whey (WP_{L_filt}) at low protein dry weight concentrations (0.4% wt/wt) were exposed to microsecond pulsed electric field (μsPEF) treatments at EF intensities between 1.25 and 12.5 kV/cm, pulse repetition frequencies between 0.38 and 85 Hz, and pulse lengths set to 10 or 100 μs. Protein aggregations were quantified spectroscopically. We report here that aggregates formed at lower temperatures for μsPEF compared with purely thermal treatments in identical treatment geometries at similar time-temperature profiles. We suggest that the observed increase in absorbance is linked to protein migration, the isoelectric point, local deprotonation phenomena of thiol groups, and cation precipitation. The μsPEF treatment time, which is dependent on the pulse repetition frequency, pulse length, and time of process, is the main driver of the increase in absorbance. High EF intensities balanced with shorter pulse repetition frequencies to ensure similar energy inputs resulted in no aggregate formation. For WP_{L_filt}, 12.5 kV/cm, 10 μs, 0.38 Hz (620 ± 96 kJ/kg; ± standard deviation) did not result in an increase in absorbance, whereas 1.25 kV/cm, 10 μs, 50 Hz (634 ± 57 kJ/kg) with similar time-temperature profiles increased the absorbance at a wavelength of 380 nm by a factor of 8.2 ± 1.7 compared with untreated WP_{L_filt}. In conclusion, the treatment time seems to dominate over high EF intensities at similar energy inputs for aggregate formation and increase in absorbance.