Combination Of Pulsed Electric Fields Research Articles

Bacterial resistance after pulsed electric fields depending on the treatment medium pH

The objective was to evaluate and compare the pulsed electric field (PEF) resistance of four Gram-positive ( Bacillus subtilis, Listeria monocytogenes, Lactobacillus plantarum, Staphylococcus aureus) and four Gram-negative ( Escherichia coli, E. coli O157:H7, Salmonella serotype Senftenberg 775W, Yersinia enterocolitica) bacterial strains under the same treatment conditions. Microbial characteristics such as cell size, shape or type of the cell envelopes did not exert the expected influence on microbial PEF resistance. The most PEF resistant bacteria depended on the treatment medium pH. For instance, L. monocytogenes, which showed the highest PEF resistance at pH 7.0, was one of the most sensitive at pH 4.0. The most PEF resistant strains at pH 4.0 were the Gram-negatives E. coli O157:H7 and S. Senftenberg. A subsequent holding of PEF-treated cells in pH 4.0 for 2 h increased the degree of inactivation up to 4 extra Log 10 cycles depending on the bacterial strain investigated. Under these treatment conditions, the most PEF resistant bacterial strains were still the pathogens S. Senftenberg and E. coli O157:H7. Industrial relevance The design of appropriate food preservation processes by PEF requires the selection of an adequate target bacterial strain, which should correspond to the most PEF resistant microorganism contaminating food. This study indicates that the pH of the treatment medium plays an important role in determining this target bacterial strain. On the other hand, the combination of PEF and subsequent holding under acidic conditions has been proven to be an effective method in order to achieve a higher level of microbial inactivation.

PULSED ELECTRIC FIELD TREATMENT OF LIQUID WHOLE EGG INOCULATED WITH SALMONELLA ENTERITIDIS

ABSTRACT The effects of pulsed electric fields (PEF) on the inactivation of Salmonella Enteritidis inoculated into liquid whole egg (LWE) and on the physical properties and the shelf‐life of LWE were studied. PEF processing conditions were 1.2 mL/s flow rate, 200 pps frequency, 2.12 μs pulse duration, 25 kV/cm electric field strength, and 250 μs total treatment time. The PEF processing caused up to 1 log10 cfu/mL reduction in S. Enteritidis population in LWE. The PEF‐treated samples were subjected to heat at 55C for 3.5 min to inactivate the remaining bacteria without denaturing the LWE. The combination of PEF and heat treatments led to a 4.3 log10 cfu/mL reduction in S. Enteritidis population (P < 0.05) and caused no significant change in viscosity, electrical conductivity, color, pH, and Brix, relative to control samples (P > 0.05). The PEF+55C treated LWE samples presented significantly longer shelf‐life at 4C compared with the control and heat treated samples (P < 0.05).

A model for evaluating therapeutic response of combined cancer treatment modalities: applied to treatment of subcutaneously implanted brain tumors (N32 and N29) in Fischer rats with pulsed electric fields (PEF) and 60Co-gamma radiation (RT).

The aim of the present study is to develop a mathematical model for evaluating therapeutic response of combined treatment modalities. The study was performed in rats of the Fischer-344 strain with rat glioma N32 or N29 tumors implanted subcutaneously on the thigh of the hind leg. Pulsed electric fields, PEF, with 16 exponentially decaying pulses with a maximum electric field strength of 140 V/mm and t(1/e)= 1 ms were first applied to the tumors. Then within 5 min radiation therapy with (60)Co-gamma radiation, RT, was given in daily fractions of 5 Gy. The animals were arranged into one group of controls and 3 groups of different kind of treatments: PEF only, RT only or combination of PEF + RT. At about 4 weeks after inoculation, the tumors were given the treatment sessions during one week. In 2 experimental series with totally 52 rats with N32 tumors, of which 16 were controls, were given 4 sessions of PEF treatments and RT (totally 20 Gy). In a special experimental series with totally 56 rats with N32 tumors, of which 10 were controls, the different groups were given 1, 2, 3 or 4 treatment sessions respectively. Another strain of glioma tumor, N29 with 62 tumors of which 14 were controls was studied in 2 series given 4PEF + 4RT and 2PEF + 4RT respectively. Fitting the data obtained from consecutive measurements of tumor volume (TV) of each individual tumor to an exponential model TV = TV(0). exp[TGR.t] estimated the tumor growth rate (TGR % per day) after the first day of treatment (t = 0). The TGR of N32 tumors treated with the combination of 4PEF + 4RT are significantly decreased compared to the controls (p < 0.0001), compared to RT alone (p < 0.0001) and compared to PEF alone (p < 0.001). The combined treatment of N32 gives significant effect on the tumor growth rate after 2, 3 and 4 treatment session while RT alone seems to be most efficient after one treatment of 5 Gy and PEF alone is most efficient after 2 treatments at 2 consecutive days. The TGR of N29 tumors treated with the combination of 4PEF + 4RT are significantly decreased compared to the controls (p < 0.05) but the combination of 2PEF + 4RT was more effective (p < 0.005). The specific therapeutic effect STE is defined as the difference between the average tumor growth rates of controls and exposed tumors divided by the average tumor growth rate of the controls. With 4PEF treatments alone the average STE value was 0.32 for N32 tumors and 0 for N29; for 4RT alone the STE values were 0.29 and 0.42 respectively, and for combined treatments 4PEF + 4RT 0.67 and 0.17 respectively. For the N29 tumors treated with 2PEF + 4RT the STE value was 0.53. The therapeutic enhancement ratio, TER, value increase with the number of treatment sessions and the TER of the combined treatments is above 1 in two of the N32 series, which indicates a synergistic effect of 4PEF + 4RT. This work demonstrate the use of our model for analyzing the combination PEF + RT, but it can also be used for evaluation the therapeutic effects of combining RT with chemotherapy or immunogenetic therapy.

Effects of pulsed electric fields on inactivation and metabolic activity of Lactobacillus plantarum in model beer.

Inactivation and sublethal injury of Lactobacillus plantarum at different pulsed electric field (PEF) strengths and total energy inputs were investigated to differentiate reversible and irreversible impacts on cell functionality. Lactobacillus plantarum was treated with PEF in model beer (MB) to determine critical values of field strength and energy input for cell inactivation. Below critical values, metabolic activity and membrane integrity were initially reduced without loss of viability. Above critical values, however, irreversible cell damage occurred. Presence of nisin or hop extract, during PEF treatment, resulted in an additional reduction of cell viability by 1;5 log cycles. Also, addition of the hop extract resulted in an additional two log cycles of sublethal injury. Partial reversibility of membrane damage was observed using propidium iodide (PI) uptake and staining. Inoculated MB containing hops was stored after PEF to evaluate the efficacy of such treatment for beer preservation. Cells were inactivated only above critical values of 13 kV x cm(-1) and 64 kJ x kg(-1); below these values cell damage was reversible. Storage experiments revealed that surviving cells were killed after 15 h storage in MB containing hops. Both reversible and irreversible cell damage due to PEF treatment was detected, depending on specific treatment conditions. The combination of PEF and hop addition is a promising nonthermal method of preservation for beer.

Reduction in Levels of Escherichia coli O157:H7 in Apple Cider by Pulsed Electric Fields

Many studies have demonstrated that high voltage pulsed electric field (PEF) treatment has lethal effects on microorganisms including Escherichia coli O157:H7; however, the survival of this pathogen through the PEF treatment is not fully understood. Fresh apple cider samples inoculated with E. coli O157:H7 strain EC920026 were treated with 10, 20, and 30 instant charge reversal pulses at electric field strengths of 60, 70, and 80 kV/cm, at 20, 30, and 42°C. To accurately evaluate the lethality of apple cider processing steps, counts were determined on tryptic soy agar (TSA) and sorbitol MacConkey agar (SMA) to estimate the number of injured and uninjured E. coli O157:H7 cells after PEF treatment. Cell death increased significantly with increased temperatures and electric field strengths. A maximum of 5.35-log10 CFU/ml (P < 0.05) reduction in cell population was achieved in samples treated with 30 pulses and 80 kV/cm at 42°C. Cell injury measured by the difference between TSA and SMA counts was found to be insignificant (P > 0.05). Under extreme conditions, a 5.91-log10 CFU/ml reduction in cell population was accomplished when treating samples with 10 pulses and 90 kV/cm at 42°C. PEF treatment, when combined with the addition of cinnamon or nisin, triggered cell death, resulting in a reduction in E. coli O157:H7 count of 6 to 8 log10 CFU/ml. Overall, the combination of PEF and heat treatment was demonstrated to be an effective pasteurization technique by sufficiently reducing the number of viable E. coli O157:H7 cells in fresh apple cider to meet U.S. Federal Drug Administration recommendations.

Effect of electric field pulses on microstructure of muscle foods and roes

The effect of pulsed electric fields (PEF) and combination of PEF and high pressure were studied on microstructure of salmon, chicken and lumpfish roes. The results showed that PEF treatment with low field strength (less than 2 kV/cm and 20–40 pulses) had considerable effect on the microstructure, i.e. the muscle cells decreased in size and gaping occurred. PEF treatment had greater effect on salmon than chicken samples. However, roes seem to tolerate up to 18.6 kV/cm and seven pulses without visible effect. Reduction of microorganisms was still not sufficient at the same time. Combination of PEF and high pressure (200–300 MPa) had more detrimental effect on microstructure than PEF treatment alone.

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 Listeria 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.

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 μs, 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°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.