Inactivation of Listeria innocua in skim milk by pulsed electric fields and nisin

Pulsed electric fields (PEF) is an emerging nonthermal processing technology used to inactivate microorganisms in liquid foods such as milk. PEF results in loss of cell membrane functionality that leads to inactivation of the microorganism. There are many processes that aid in the stability and safety of foods. The combination of different preservation factors, such as nisin and PEF, to control microorganisms should be explored. The objective of this research was to study the inactivation of Listeria innocua suspended in skim milk by PEF as well as the sensitization of PEF treated L. innocua to nisin. The selected electric field intensity was 30, 40 and 50 kV/cm and the number of pulses applied was 10.6, 21.3 and 32. The sensitization exhibited by PEF treated L. innocua to nisin was assessed for 10 or 100 IU nisin/ml. A progressive decrease in the population of L. innocua was observed for the selected field intensities, with the greatest reduction being 2 1/2 log cycles (U). The exposure of L. innocua to nisin after PEF had an additive effect on the inactivation of the microorganism as that exhibited by the PEF alone. As the electric field, number of pulses and nisin concentration increased, synergism was observed in the inactivation of L. innocua as a result of exposure to nisin after PEF. The reduction of L. innocua accomplished by exposure to 10 IU nisin/ml after 32 pulsed electric fields was 2, 2.7, and 3.4 U for an electric field intensity of 30, 40, and 50 kV/cm, respectively. Population of L. innocua subjected to 100 IU nisin/ml after PEF was 2.8-3.8 U for the selected electric field intensities and 32 pulses. The designed model for the inactivation of L. innocua as a result of the PEF followed by exposure to nisin proved to be accurate in the prediction of the inactivation of L. innocua in skim milk containing 1.2 or 37 IU nisin/ml. Inactivation of L. innocua in skim milk containing 37 IU nisin/ml resulted in a decrease in population of 3.7 U.     

Non-thermal pasteurization of fruit juices by combining high-intensity pulsed electric fields with natural antimicrobials

The effect of high-intensity pulsed electric fields (HIPEF) on the Salmonella Enteritidis and Escherichia coli O157:H7 populations inoculated in apple, pear, orange and strawberry juices as influenced by treatment time and pulse frequency was investigated. Combinations of HIPEF (35 kV/cm, 4 μs pulse length in bipolar mode without exceeding 40 °C) with citric acid or cinnamon bark oil against these pathogenic microorganisms in fruit juices were also evaluated. Treatment time was the more influential factor on the microbial reduction in all the fruit juices analyzed. S. Enteritidis and E. coli O157:H7 were reduced by more than 5.0 log₁₀ units in orange juice treated by only HIPEF; whereas strawberry, apple and pear juices were pasteurized when HIPEF was combined with citric acid at 0.5, 1.5, 1.5%, respectively, or cinnamon bark oil at 0.05, 0.1 and 0.1%, respectively. Synergistic and additive killing effects against S. Enteritidis and E. coli O157:H7 in fruit juices by combining treatments were observed.

Industrial relevance

The use of high-intensity pulsed electric fields treatment as a non-thermal pasteurization method in combination with organic acids or essential oils is an effective process for eliminating S. Enteritidis and E. coli O157:H7 populations in fruit juices upper 5.0 log₁₀ reductions. Therefore, combinations of those treatments may help to ensure the microbiological safety in juice products, and to reduce the risk of food-borne illness caused by the consumption of these kinds of foods.     

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.     

Combined effect of nisin and pulsed electric fields on the inactivation of Escherichia coli

The Doehlert design was applied in order to investigate the combined effect of nisin and high voltage pulsed electric fields (PEF) on the inactivation of Escherichia coli in simulated milk ultrafiltrate media. Nisin alone was totally inactivated by PEF, but in the presence of bacterial cells a protective effect was observed. However, the effectiveness of nisin was still decreased when bacterial cells were subjected to the combined treatment. In spite of this phenomenon, an almost additive response emerged as a consequence of the combined treatment. A 4-log cycle reduction may be accomplished with around 1,000 IU/ml (7.15 microM) of nisin and three pulses of 11.25 kV/cm or 500 IU/ml for five pulses of the same intensity. The observed efficacy arising from the combination of both treatments suggests the possibility of using PEF for improving the action spectrum of natural antimicrobials.     

Transmission electron microscopy of Listeria innocua treated by pulsed electric fields and nisin in skimmed milk

Pulsed electric fields (PEF) is a nonthermal food preservation process where organoleptic and nutritional properties of the food are maintained. PEF is known to inactivate microorganisms by causing dielectric breakdown of the cell membrane, thus altering the functionality of the membrane as a semipermeable barrier. The extent of damage of the cell membrane, whether visible in the form of a pore or as loss of membrane functionality leads to the inactivation of the microorganism. The objective of this study was to investigate under transmission electron microscopy (TEM) the morphological changes on Listerit innocua as a result of PEF treatment in skimmed milk containing nisin. L. innocua was subjected to PEF at selected electric field intensities of 30, 40, and 50 kV/cm. L. innocua was treated by PEF in both skimmed milk with and without 37 IU nisin/ml. L. innocua treated by PEF in skimmed milk exhibited an increase in the cell wall roughness. cytoplasmic clumping, leakage of cellular material, and rupture of the cell walls and cell membranes. L. innocua subjected to PEF in skimmed milk containing 37 IU nisin/ml exhibited an increased cell wall width. At the highest electric field intensity, 50 kV/cm, elongation of the cell length was observed. There were no morphological differences between cells treated by PEF in skimmed milk with or without nisin. The combination of PEF and nisin exhibit an additive effect in the morphological damage observed on L. innocua. Pore formation was observed on L. innocua for an electric field intensity of 40 kV/cm. The inactivation of L. innocua was a consequence of rupture of the cell membrane and loss of cell membrane functionality.     

High-intensity pulsed electric field variables affecting Staphylococcus aureus inoculated in milk

Staphylococcus aureus is an important milk-related pathogen that is inactivated by high-intensity pulsed electric fields (HIPEF). In this study, inactivation of Staph. aureus suspended in milk by HIPEF was studied using a response surface methodology, in which electric field intensity, pulse number, pulse width, pulse polarity, and the fat content of milk were the controlled variables. It was found that the fat content of milk did not significantly affect the microbial inactivation of Staph. aureus. A maximum value of 4.5 log reductions was obtained by applying 150 bipolar pulses of 8 μs each at 35 kV/cm. Bipolar pulses were more effective than those applied in the monopolar mode. An increase in electric field intensity, pulse number, or pulse width resulted in a drop in the survival fraction of Staph. aureus. Pulse widths close to 6.7 μs lead to greater microbial death with a minimum number of applied pulses. At a constant treatment time, a greater number of shorter pulses achieved better inactivation than those treatments performed at a lower number of longer pulses. The combined action of pulse number and electric field intensity followed a similar pattern, indicating that the same fraction of microbial death can be reached with different combinations of the variables. The behavior and relationship among the electrical variables suggest that the energy input of HIPEF processing might be optimized without decreasing the microbial death.     

Inactivation of Salmonella Typhimurium and Staphylococcus aureus by pulsed electric fields in liquid whole egg

In this study, the lethal effectiveness of pulsed electric fields (PEF) on the inactivation of Salmonella enterica subs. enterica ser. Typhimurium and Staphylococcus aureus in liquid whole egg (LWE) has been investigated. Maximum inactivation levels of 4 and 3 Log₁₀ cycles of the population of Salmonella Typhimurium and S. aureus were achieved with treatments of 45 kV/cm, 30 μs and 419 kJ/kg, and 40 kV/cm for 15 μs and 166 kJ/kg, respectively. The non-linear kinetics of inactivation observed for both microorganisms at all the investigated electric field strengths were described by mathematical equations based on the Weibull distribution. The developed equations enabled to compare the microbial resistance to PEF and to establish the most suitable treatment conditions to achieve a determined level of microbial inactivation. PEF treatments varying from 30 kV/cm, 67 µs and 393 kJ/kg to 45 kV/cm, 19 µs and 285 kJ/kg allow to reduce 3 Log₁₀ cycles the population of the microorganism of concern in PEF food processing of LWE, Salmonella Typhimurium.Industrial relevanceThe data presented in this investigation in terms of electric field strength, specific energy and treatment time result of relevance to evaluate the possibilities of PEF technology to pasteurize LWE with this technology. The models developed in this study can be applied to engineering design, and for the evaluation and optimization of the PEF technology as a new technique to obtain Salmonella free LWE.Based on our results it is not recommended to apply treatments of energy levels higher than 250 kJ/kg, since PEF lethality hardly increased but markedly augmented the energetic costs. For these energy values, PEF technology by itself is not sufficient (3 Log10 cycles in the best case scenario) to assure the safe security of LWE. Therefore, intelligent combinations of PEF with other preservation technologies have to be developed in order to use pulsed electric fields as an alternative to heat pasteurization of LWE.     

Enhancing the lethal effect of high-intensity pulsed electric field in milk by antimicrobial compounds as combined hurdles

High-intensity pulsed electric field (HIPEF) is a nonthermal treatment studied for its wide antimicrobial spectrum on liquid food, including milk and dairy products. Moreover, the antimicrobial effect of HIPEF may be enhanced by combining HIPEF with other treatments as hurdles. Nisin and lysozyme are natural antimicrobial compounds that could be used in combination with HIPEF. Therefore, the purpose of this study was to determine the effect of combining HIPEF with the addition of nisin and lysozyme to milk inoculated with Staphylococcus aureus with regard to different process variables. The individual addition of nisin and lysozyme did not produce any reduction in cell population within the proposed range of concentrations, whereas their combination resulted in a pH-dependent microbial death of Staph. aureus. The addition of nisin and lysozyme to milk combined with HIPEF treatment resulted in a synergistic effect. Applying a 1,200-μs HIPEF treatment time to milk at pH 6.8 containing 1 IU/mL of nisin and 300 IU/mL of lysozyme resulted in a reduction of more than 6.2 log units of Staph. aureus. Final counts resulting from the addition of nisin and lysozyme and applying HIPEF strongly depended on both the sequence of application and the milk pH. Thus, more research is needed to elucidate the mode of action of synergism as well as the role of different process variables, although the use of HIPEF in combination with antimicrobial compounds such as nisin and lysozyme is shown to be potentially useful in processing milk and dairy products.     

Combined effect of temperature and pulsed electric fields on soya milk lipoxygenase inactivation

Pulsed electric fields (PEF) were applied to freshly prepared soya milk using a laboratory scale continuous PEF system to study the feasibility of inactivating lipoxygenase (LOX). Square wave PEF using different combinations of pre-treatment temperature, electric field strength and treatment time were evaluated in this study. Inactivation curves for the enzyme were plotted for each parameter and inactivation kinetics were calculated and modelled. Results showed the highest level of inactivation (84.5%) was obtained using a combination of preheating to 50 °C, and a PEF treatment time of 100 μs at 40 kV/cm. Inactivation of LOX activity as a function of treatment time could be described using a first order kinetic model. Calculated D values following pre-heating to 50 °C were 172.9, 141.6 and 126.1 μs at 20, 30 and 40 kV/cm, respectively.     

Component release after exposure of Staphylococcus aureus cells to pulsed electric fields

The objective of this work was to get further insights on the mechanism of inactivation of bacterial cells by pulsed electric fields (PEF) through the study of the release of intracellular components after exposing Staphylococcus aureus cells in McIvlaine buffer (pH 7.0, 2 mS/cm) to PEF treatments of different intensity (18 and 25 kV/cm) and treatment times (from 20 to 400 μs). Release of most compounds, except proteins, was almost immediate after the treatment, but the relative amount released depended on the molecule studied. A good correlation between the release of the smallest components studied (particularly ions) and membrane permeabilization (as measured by NaCl sensitization and PI entry) was observed. On the other hand, results obtained suggested that S. aureus inactivation by PEF would be related to the exit of cytoplasmic proteins of a molecular weight higher than 6 kDa. Results obtained in this work indicated that increasing PEF treatment time would reduce the capability of S. aureus cells to repair the electropores formed and suggested that this might be due to the formation of pores of a larger size, which S. aureus cells would be unable to reseal in a situation of homeostasis loss.Industrial relevanceResults reported here can help to design more effective treatments for microbial inactivation using PEF on food, and therefore facilitate its industrial implementation.     

Microbial inactivation and shelf life comparison of 'cold' hurdle processing with pulsed electric fields and microfiltration, and conventional thermal pasteurisation in skim milk

Thermal pasteurisation (TP) is the established food technology for commercial processing of milk. However, degradation of valuable nutrients in milk and its sensory characteristics occurs during TP due to substantial heat exposure. Pulsed electric fields (PEF) and microfiltration (MF) both represent emerging food processing technologies allowing gentle milk preservation at lower temperatures and shorter treatment times for similar, or better, microbial inactivation and shelf stability when applied in a hurdle approach compared to TP. Incubated raw milk was used as an inoculum for the enrichment of skim milk with native microorganisms before PEF, MF, and TP processing. Inoculated milk was PEF-processed at electric field strengths between 16 and 42 kV/cm for treatment times from 612 to 2105 μs; accounting for energy densities between 407 and 815 kJ/L, while MF was applied with a transmembrane flux of 660 L/h m². Milk was TP-treated at 75 °C for 24 s. Comparing PEF, MF, and TP for the reduction of the native microbial load in milk led to a 4.6 log₁₀ CFU/mL reduction in count for TP, which was similar to 3.7 log₁₀ CFU/mL obtained by MF (P ≥ 0.05), and more effective than the 2.5 log₁₀ CFU/mL inactivation achieved by PEF inactivation (at 815 kJ/L (P < 0.05)). Combined processing with MF followed by PEF (MF/PEF) produced a 4.1 (at 407 and 632 kJ/L), 4.4 (at 668 kJ/L) and 4.8 (at 815 kJ/L) log₁₀ CFU/mL reduction in count of the milk microorganisms, which was comparable to that of TP (P ≥ 0.05). Reversed processing (PEF/MF) achieved comparable reductions of 4.9, 5.3 and 5.7 log₁₀ CFU/mL (at 407, 632 and 668 kJ/L, respectively (P ≥ 0.05)) and a higher inactivation of 7.1 log₁₀ (at 815 kJ/mL (P < 0.05)) in milk than for TP. Microbial shelf life of PEF/MF-treated (815 kJ/L) and TP-treated milk stored at 4 °C was analysed over 35 days for total aerobic; enterobacteria; yeasts and moulds; lactobacilli; psychrotroph; thermoduric psychrotroph, mesophilic, and thermophilic; and staphylococci counts. For both PEF/MF and TP-treated milk an overall shelf stability of 7 days was observed based on total aerobic counts (P ≥ 0.05). Milk hurdle processing with PEF/MF at its most effective treatment parameters produced greater microbial inactivation and overall similar shelf stability at lower processing temperatures compared to TP. With higher field strength, shorter treatment time, larger energy density, and rising temperature the efficacy of PEF/MF increased contrary to MF/PEF. Thus, PEF/MF represents a potential alternative for ‘cold’ pasteurisation of milk with improved quality.     

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.     

Inactivation of Listeria innocua in liquid whole egg by pulsed electric fields and nisin

Consumer demand for fresh-like products with little or no degradation of nutritional and organoleptic properties has led to the study of new technologies in food preservation. Pulsed electric fields (PEF) is a nonthermal preservation method used to inactivate microorganisms mainly in liquid foods. Microorganisms in the presence of PEF suffer cell membrane damage. Nisin is a natural antimicrobial known to disrupt cell membrane integrity. Thus the combination of PEF and nisin represents a hurdle for the survival of Listeria innocua in liquid whole egg (LWE). L. innocua suspended in LWE was subjected to two different treatments: PEF and PEF followed by exposure to nisin. The selected frequency and pulse duration for PEF was 3.5 Hz and 2 micros, respectively. Electric field intensities of 30, 40 and 50 kV/cm were used. The number of pulses applied to the LWE was 10.6, 21.3 and 32. The highest extent of microbial inactivation with PEF was 3.5 log cycles (U) for an electric field intensity of 50 kV/cm and 32 pulses. Treatment of LWE by PEF was conducted at low temperatures, 36 degrees C being the highest. Exposure of L. innocua to nisin following the PEF treatment exhibited an additive effect on the inactivation of the microorganism. Moreover, a synergistic effect was observed as the electric field intensity, number of pulses and nisin concentration increased. L. innocua exposed to 10 IU nisin/ml after PEF exhibited a decrease in population of 4.1 U for an electric field intensity of 50 kV/cm and 32 pulses. Exposure of L. innocua to 100 IU nisin/ml following PEF resulted in 5.5 U for an electric field intensity of 50 kV/cm and 32 pulses. The model developed for the inactivation of L. innocua by PEF and followed by exposure to nisin proved to be accurate (p = 0.05) when used to model the inactivation of the microorganism by PEF in LWE with 1.2 or 37 IU nisin/ml. The presence of 37 IU nisin/ml in LWE during the PEF treatment for an electric field intensity of 50 kV/cm and 32 pulses resulted in a decrease in the population of L. innocua of 4.4 U.     

Inactivation of Salmonella enterica Ser. Enteritidis in tomato juice by combining of high-intensity pulsed electric fields with natural antimicrobials

ABSTRACT:  The effect of high-intensity pulsed electric field (HIPEF) treatment (35kV/cm, 4 μs pulse length in bipolar mode without exceeding 38 °C) as influenced by treatment time (200, 600, and 1000 μs) and pulse frequency (100, 150, and 200 Hz) for inactivating Salmonella enterica ser. Enteritidis inoculated in tomato juice was evaluated. Similarly, the effect of combining HIPEF treatment with citric acid (0.5%, 1.0%, 1.5%, and 2.0%[wt/vol]) or cinnamon bark oil (0.05%, 0.10%, 0.2%, and 0.3%[vol/vol]) as natural antimicrobials against S. Enteritidis in tomato juice was also studied. Higher treatment time and lower pulse frequency produced the greater microbial inactivation. Maximum inactivation of S. Enteritidis (4.184 log₁₀ units) in tomato juice by HIPEF was achieved when 1000 μs and 100 Hz of treatment time and pulse frequency, respectively, were applied. However, a greater microbial inactivation was found when S. Enteritidis was previously exposed to citric acid or cinnamon bark oil for 1 h in tomato juice. Synergistic effects were observed in HIPEF and natural antimicrobials. Nevertheless, combinations of HIPEF treatment with 2.0% of citric acid or 0.1% of cinnamon bark oil were needed for inactivating S. Enteritidis by more than 5.0 log₁₀ units (5.08 and 6.04 log₁₀ reductions, respectively). Therefore, combinations of HIPEF with organic acids or essential oils seem to be a promising method to achieve the pasteurization in these kinds of products.     

Alkaline phosphatase and microbial inactivation by pulsed electric field in bovine milk

The effects of pulsed electric field (PEF) treatments at field intensities of 25–37 kV cm^− 1 and final PEF treatment temperatures of 15 °C and 60 °C on the inactivation of alkaline phosphatase (ALP), Total Plate Count (TPC), Pseudomonas and Enterobacteriaceae counts were determined in raw skim milk. At 15 °C, PEF treatments of 28 to 37 kV cm^− 1 resulted in 24–42% inactivation in ALP activity and < 1 log reduction in TPC and Pseudomonas count, while the Enterobacteriaceae count was reduced by at least 2.1 log units to below the detection limit of 1 CFU mL^− 1. PEF treatments of 25 to 35 kV cm^− 1 at 60 °C resulted in 29–67% inactivation in ALP activity and up to 2.4 log reduction in TPC, while the Pseudomonas and Enterobacteriaceae counts were reduced by at least 5.9 and 2.1 logs, respectively, to below the detection limit of 1 CFU mL^− 1. Kinetic studies suggested that the effect of field intensity on ALP inactivation at the final PEF treatment temperature of 60 °C was more than twice that at 15 °C. A combined effect was observed between the field intensity and temperature in the inactivation of both ALP enzyme and the natural microbial flora in raw skim milk.Industrial relevanceMilk has been pasteurised to ensure its safety and extend its shelf life. However, the need for retaining heat-sensitive nutrient and sensory properties of milk has resulted in interest in the application of alternative technologies. The results of the current study suggest that PEF as a non-thermal process can be employed for the treatment of raw milk in mild temperature to achieve adequate safety and shelf life while preserving the heat-sensitive enzymes, nutrients and bioactive compounds.     

Factors affecting the inactivation of the natural microbiota of milk processed by pulsed electric fields and cross-flow microfiltration

Prior to processing milk and cream were standardised and homogenised. Skim milk was cross-flow microfiltered (CFMF) prior to treatment with pulsed electric fields (PEF) or high temperature short time (HTST) pasteurization. The effect of temperature of the skim milk and product composition on the efficacy of PEF treatment was determined. The electrical conductivity of the product was related to fat and solids content and increased 5% for every g/kg increase of solids and decreased by nearly 0·7% for every g/kg increase of fat. From the three microbial groups analyzed (mesophilic, coliform, and psychrotroph) in milks differences (P<0·05) in the inactivation of mesophilic microorganisms were observed between the counts following PEF treatment, while HTST pasteurization resulted in higher reductions in all different counts than those obtained after PEF. Increasing the skim milk temperature prior to PEF treatment to about 34°C showed equivalent reductions in microbial counts to skim milk treated at 6°C in half the time. The reductions achieved by a combination of CFMF and PEF treatments were comparable to those achieved when CFMF was combined with HTST pasteurization. A higher reduction in coliform counts was observed in homogenised products subjected to PEF than in products that were only standardised for fat content.     

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.     

Color and viscosity of watermelon juice treated by high-intensity pulsed electric fields or heat

The effects of high-intensity pulsed electric field (HIPEF) processing (35 kV/cm for 1727 μs applying 4-μs pulses at 188 Hz in bipolar mode) on color, viscosity and related enzymes in watermelon juice were evaluated during 56 days of storage and compared to thermal treatments (90 °C for 60 s or 30 s). HIPEF-treated juice maintained brighter red color than thermally treated juices along the storage time. In addition, the application of HIPEF as well as heat at 90 °C for 60 s led to juices with higher viscosity than those untreated for 56 days of storage. On the other hand, peroxidase (POD) was inactivated more efficiently after HIPEF processing than after applying heat treatments. However, the thermally processed juice at 90 °C for 60 s kept the lowest residual POD activity values beyond day 7 of storage. Differences in lipoxygenase (LOX) activity among treatments were not appreciated at day 0. However, storage time had a strong reducing influence on the enzyme activity of heat-treated samples. A substantial loss of pectin methylesterase (PME) activity (more than 50%) was observed in all the treated juices, whereas a slight reduction in polygalacturonase (PG) activity was only achieved after the HIPEF treatment. The use of HIPEF technology could be an alternative to thermal treatments and could contribute to better maintain valuable attributes of watermelon juice.Industrial RelevanceHIPEF processing is a feasible alternative to thermal treatments to obtain watermelon juice, achieving optimal inactivation of deleterious microorganisms and quality-related enzymes. HIPEF-treated watermelon juices exhibit better physical properties such as color or viscosity than thermally treated juices throughout storage. Thus, HIPEF technology can help processors to obtain juices that keep their fresh characteristics, thus being better accepted by consumers.     

Effects of combined exposure of micrococcus luteus to nisin and pulsed electric fields

Death and injury following exposure of Micrococcus luteus to nisin and pulsed electric field (PEF) treatment were investigated in phosphate buffer (pH 6.8, σ=4.8 ms/cm at 20°C). Four types of experiment were carried out, a single treatment with nisin (100 IU/ml at 20°C for 2 h), a single PEF treatment, a PEF treatment followed by incubation with nisin (as before) and addition of nisin to the bacterial suspension prior to the PEF treatment. The application of nisin clearly enhanced the lethal effect of PEF treatment. The bactericidal effect of nisin reduced viable counts by 1.4 log₁₀ units. Treatment with PEF (50 pulses at 33 kV/cm) resulted in a reduction of 2.4 log₁₀ units. PEF treatment followed by nisin caused a reduction of 5.2 log₁₀ units in comparison with a 4.9 log₁₀ units reduction obtained with nisin followed by PEF. Injury of surviving cells was investigated using media with different concentrations of salt. Sublethally damaged cells of M. luteus could not be detected by this means, following PEF treatment.