In this study conducted at Food Science Australia (FSA) and Berlin University of Technology (BUT), the effects of pulsed electric field (PEF) treatment, a novel non-thermal processing technology on bovine milk microflora and native enzymes and on the rheological and textural properties of rennet-induced milk gels was investigated. The PEF treatments were conducted at field intensities of 25-37 kV cm-1 (up to 50 kV cm-1)and temperature range of 30°C to 75ºC. Native milk enzymes selected for the study included alkaline phosphatase, lipase, xanthine oxidase and plasminand microbiological study included determining Total Plate Count (TPC) and Pseudomonas and Enterobacteriaceae counts in skim milk. At 30ºC PEF treatment at maximum field intensity inactivated AlP by 42% while at 60oC inactivation was higher (67%). Under these treatment conditions less than1 log reduction in TPC and Pseudomonas count and 2.1 logs reduction in the Enterobacteriaceae count was achieved at 30oC while at 60ºC TPC dropped by 2.4 logs and Pseudomonas and Enterobacteriaceae counts were reduced by 5.9 and 2.1 logs, respectively to below the detection limit of 1 CFU mL-1. Combining PEF treatment with heat increased the inactivation level of all enzymes which showed an increasing trend with increasing field intensity and temperature. Treatment time (4.8, 9.6, 19.2, 28.8 and 38.4 µs) was controlled by either changing the pulse frequencies (100-400 Hz) or product flow rate (30-240 mL min-1) at a constant field intensity of 31 kV cm-1 and it was found that changing the flow rate was a more effective way of enzyme inactivation than changing the frequency due to longer exposure time of enzymes to heat and field intensity. The size of casein micelles and fat globules was not affected by PEF treatment while severe heating of milk at 97oC for 10 min decreased both micelle and fat globule sizes marginally. The coagulation time of rennet-induced gels made from PEF-treated (35 to 50 kV cm-1) milks (whole and skim) increased as the treatment intensity increased, but remained shorter than gels made from pasteurised milk. The PEF treatment of milk at various field intensities and temperatures adversely affected the G′, G′′ and firmness of gels, but the effects were less pronounced than in gels made from pasteurised milks. This study concludes that for successful application in milk processing the PEF treatment needs to be combined with mild heat treatment. This approach could achieve safer milk with less damage to milk functionality. However, the quest for a suitable quality assurance indicator enzyme will need more extensive studies.

The PEF treatments were conducted at field intensities of 25-37 kV cm-1 (up to 50 kV cm-1)and temperature range of 30°C to 75ºC. Native milk enzymes selected for the study included alkaline phosphatase, lipase, xanthine oxidase and plasminand microbiological study included determining Total Plate Count (TPC) and Pseudomonas and Enterobacteriaceae counts in skim milk. At 30ºC PEF treatment at maximum field intensity inactivated AlP by 42% while at 60oC inactivation was higher (67%). Under these treatment conditions less than1 log reduction in TPC and Pseudomonas count and 2.1 logs reduction in the Enterobacteriaceae count was achieved at 30oC while at 60ºC TPC dropped by 2.4 logs and Pseudomonas and Enterobacteriaceae counts were reduced by 5.9 and 2.1 logs, respectively to below the detection limit of 1 CFU mL-1. Combining PEF treatment with heat increased the inactivation level of all enzymes which showed an increasing trend with increasing field intensity and temperature. Treatment time (4.8, 9.6, 19.2, 28.8 and 38.4 µs) was controlled by either changing the pulse frequencies (100-400 Hz) or product flow rate (30-240 mL min-1) at a constant field intensity of 31 kV cm-1 and it was found that changing the flow rate was a more effective way of enzyme inactivation than changing the frequency due to longer exposure time of enzymes to heat and field intensity. The size of casein micelles and fat globules was not affected by PEF treatment while severe heating of milk at 97oC for 10 min decreased both micelle and fat globule sizes marginally. The coagulation time of rennet-induced gels made from PEF-treated (35 to 50 kV cm-1) milks (whole and skim) increased as the treatment intensity increased, but remained shorter than gels made from pasteurised milk. The PEF treatment of milk at various field intensities and temperatures adversely affected the G′, G′′ and firmness of gels, but the effects were less pronounced than in gels made from pasteurised milks. This study concludes that for successful application in milk processing the PEF treatment needs to be combined with mild heat treatment. This approach could achieve safer milk with less damage to milk functionality. However, the quest for a suitable quality assurance indicator enzyme will need more extensive studies.

Pulsed electric field (PEF) processing has been demonstrated to be an effective non-thermal pasteurization method for food-treatment applications. With this method, high voltage, short-duration pulses are applied to a chamber through which liquid food is passed. If the voltage applied and the corresponding electric field develops a potential higher than a critical trans-membrane potential, the pores expand, and the membrane of the living cell is ruptured. Due to the lower amount of energy consumed during a PEF process, the temperature of the liquid is kept much lower than as opposed to conventional pasteurization. The PEF method thus kills bacteria and other microorganisms while preserving the nutrition and taste of the liquid foods. Although the parameter responsible for inactivation is the voltage applied, for any given voltage, the conductivity of the liquid defines a current through the liquid that causes the temperature to rise. Therefore, preventing excessive heating of the liquid requires the application of an efficient waveform. According to the literature, the most efficient waveform is a square wave since the entire energy applied would be used for the inactivation process. Although some power supplies are capable of generating such a waveform, the generation of an efficient waveform that satisfies all the requirements for producing a viable product for PEF applications is still a challenging problem. In this research, a cascadable pulse generator, based on a Marx generator design, was designed and implemented in order to generate a pulsed waveform for the treatment of liquid food. IGBT switches were used to charge capacitors in parallel and to discharge them in series as a means of generating a high voltage at the output. The design was implemented and tested for two stages, generating up to 6 kV and 1.6 kA square pulses with a controllable pulse width from 1 µs to 10 µs. Up to 3 switches were connected in parallel to enhance the current capability of the system. Also investigated are ways to improve the transient time by enhancing the IGBT driver circuit. The effect of design parameters such as pulse width, voltage, and current on the temperature rise in the liquid was also studied. A variety of liquid foods with different conductivities were tested in order to confirm the functionality of the system.

Pulsed electric field (PEF) processing has been demonstrated to be an effective non-thermal pasteurization method for food-treatment applications. With this method, high voltage, short-duration pulses are applied to a chamber through which liquid food is passed. If the voltage applied and the corresponding electric field develops a potential higher than a critical trans-membrane potential, the pores expand, and the membrane of the living cell is ruptured. Due to the lower amount of energy consumed during a PEF process, the temperature of the liquid is kept much lower than as opposed to conventional pasteurization. The PEF method thus kills bacteria and other microorganisms while preserving the nutrition and taste of the liquid foods. Although the parameter responsible for inactivation is the voltage applied, for any given voltage, the conductivity of the liquid defines a current through the liquid that causes the temperature to rise. Therefore, preventing excessive heating of the liquid requires the application of an efficient waveform. According to the literature, the most efficient waveform is a square wave since the entire energy applied would be used for the inactivation process. Although some power supplies are capable of generating such a waveform, the generation of an efficient waveform that satisfies all the requirements for producing a viable product for PEF applications is still a challenging problem. In this research, a cascadable pulse generator, based on a Marx generator design, was designed and implemented in order to generate a pulsed waveform for the treatment of liquid food. IGBT switches were used to charge capacitors in parallel and to discharge them in series as a means of generating a high voltage at the output. The design was implemented and tested for two stages, generating up to 6 kV and 1.6 kA square pulses with a controllable pulse width from 1 µs to 10 µs. Up to 3 switches were connected in parallel to enhance the current capability of the system. Also investigated are ways to improve the transient time by enhancing the IGBT driver circuit. The effect of design parameters such as pulse width, voltage, and current on the temperature rise in the liquid was also studied. A variety of liquid foods with different conductivities were tested in order to confirm the functionality of the system.

L’intégration croissante des procédés innovants comme les ultrasons, les champs magnétiques, et les champs électriques pulsés a pour but d’améliorer et de stabiliser le déroulement des procédés de fermentation. Le champ électrique pulsé (CEP) est un procédé athermique généralement utilisé pour l’inactivation des pathogènes (Barbosa-Cánovas et al., 2001) ainsi que pour l’extraction des composés intracellulaires d’intérêt (El Zakhem et al., 2006a; Vorobiev & Lebovka 2006). Dans ce travail de thèse, nous proposons d’évaluer l’activité microbienne des cellules soumises à un traitement électrique modéré. Un intérêt particulier est apporté à des aspects fondamentaux comme la croissance et le métabolisme des cellules. Sur le plan technologique, le but fondamentale est de mettre en place et optimiser des protocoles de stimulation de microorganismes pour intensifier les bioprocédés. La fermentation de microorganismes stimulés par CEP a montré des cinétiques plus rapides que les levures non traitées. La stimulation de l’activité fermentaire s’est révélée grâce au suivi de la masse du milieu, les solutés solubles, l’absorbance, les sucres... L’optimisation des protocoles de stimulation a permis de réveler deux comportements logarithmique et saturé. Il a été montré une dépendance importante de l’énergie spécifique sur certains aspects physiologiques notamment la taille et le nombre de colonies. / The continually increasing integration of innovative technologies such as ultrasound, magnetic fields, and pulsed electric fields aims to improve and stabilize the course of fermentation processes. The pulsed electric field (PEF) is an athermal process generally used for pathogen inactivation (Barbosa-Canovas et al., 2001) and for the extraction of intracellular compounds of interest (El Zakhem et al., 2006a; Vorobiev & Lebovka 2006). In this thesis, we propose to evaluate the microbial activity of cells subjected to a moderate electric treatment. Special consideration is given to key aspects such as growth and cell metabolism. Technologically, the fundamental purpose is to implement and optimize microorganisms’ stimulation protocols to intensify their bioprocesses. The positive impact of PEF pre-treatment on yeast cells was shown by their faster fermentation kinetics compared to the control. This was proven by monitoring the weight of the ferment, the soluble solutes, the UV absorbance, and sugar consumption profiles. Two behaviors of electrostimulation, “logarithmic” and “saturated”, were revealed by optimization of the stimulation protocols. Finally, a relationship between the growth rate, the size of the colonies and the applied specific energy is deduced.