Alexander Mathys

Review: Potential of High Hydrostatic Pressure and Pulsed Electric Fields for Energy Efficient and Environmentally Friendly Food Processing

The application of emerging, novel processing techniques such as high hydrostatic pressure or pulsed electric fields can be utilized to replace, enhance or modify conventional techniques of food production. In addition to quality improvements and consumer benefits by gentle microbial inactivation and improvement of mass transfer processes, their potential to improve energy efficiency and sustainability of food production will be discussed within this review.

Mechanisms of endospore inactivation under high pressure

It is well known that spore germination and inactivation can be achieved within a broad temperature and pressure range. The existing literature, however, reports contradictory results concerning the effectiveness of different pressure-temperature combinations and the underlying inactivation mechanism(s). Much of the published kinetic data are prone to error as a result of unstable process conditions or an incomplete investigation of the entire inactivation pathway. Here, we review this field of research, and also discuss an inactivation mechanism of at least two steps and propose an inactivation model based on current data. Further, spore resistance properties and matrix interactions are linked to spore inactivation effectiveness.

The impact of high pressure and temperature on bacterial spores: inactivation mechanisms of Bacillus subtilis above 500 MPa.

UNLABELLED High-pressure thermal sterilization (HPTS) is an emerging technology to produce shelf stable low acid foods. Pressures below 300 MPa can induce spore germination by triggering germination receptors. Pressures above 500 MPa could directly induce a Ca+2-dipicolinic acid (DPA) release, which triggers the cortex-lytic enzymes (CLEs). It has been argued that the activated CLEs could be inactivated under HPTS conditions. To test this claim, a wild-type strain and 2 strains of Bacillus subtilis spores lacking germinant receptors and one of 2 CLEs were treated simultaneously from 550 to 700 MPa and 37 to 80 C (slow compression) and at 60 to 80 C up to 1 GPa (fast compression). Besides, an additional heat treatment to determine the amount of germinated cells, we added TbCl3 to detect the amount of DPA released from the spore core via fluorescent measurement. After pressure treatment for 120 min at 550 MPa and 37 °C, no inactivation was observed for the wild-type strain. The amount of released DPA correlated to the amount of germinated spores, but always higher compared to the belonging cell count after pressure treatment. The release of DPA and the increase of heat-sensitive spores confirm that the inactivation mechanism during HPTS passes through the physiological states: (1) dormancy, (2) activation, and (3) inactivation. As the intensity of treatment increased, inactivation of all spore strains also strongly increased (up to -5.7 log10), and we found only a slight increase in the inactivation of one of the CLE (sleB). Furthermore, above a certain threshold pressure, temperature became the dominant influence on germination rate. PRACTICAL APPLICATION   The continuous increase of high-pressure (HP) research over the last several decades has already generated an impressive number of commercially available HP pasteurized products. Furthermore, research helped to provoke the certification of a pressure-assisted thermal sterilization process by the U.S. FDA in February 2009. However, this promising sterilization technology has not yet been applied in industrial settings. An improved understanding of spore inactivation mechanisms and the ability to calculate desired inactivation levels will help to make this technology available for pilot studies and commercialization at an industrial scale. Moreover, if the synergy between pressure and elevated temperature on the inactivation rate could be identified, clarification of the underlying inactivation mechanism during HP thermal sterilization could help to further optimize the process of this emerging technology.

High pressure thermal sterilization – development and application of temperature controlled spore inactivation studies

High pressure thermal sterilization is an emerging technology that can produce uniform, minimally processed foods of high quality, better than heat treatment alone. At present, it has not yet been successfully introduced into the food industry, possibly due to the less known inactivation mechanism of high resistant bacterial spores. This study developed and used a new analytical tool to improve the understanding of spore mechanisms at high pressures and temperatures. The generated data were exemplarily incorporated into analyses of industrial sterilization processes. An improved understanding of the mechanisms of spore inactivation will aid in the food safety assessment of high pressure thermal sterilization in particular, and also assist in the commercialization of this novel process, facilitating adoption by industry.

The release of dipicolinic acid--the rate-limiting step of Bacillus endospore inactivation during the high pressure thermal sterilization process.

High pressure combined with elevated temperatures can produce low acid, commercially sterile and shelf-stable foods. Depending on the temperature and pressure levels applied, bacterial endospores pass through different pathways, which can lead to a pressure-induced germination or inactivation. Regardless of the pathway, Bacillus endospores first release pyridine-2,6-dicarboxylic acid (DPA), which contributes to the low amount of free water in the spore core and is consequently responsible for the spores high resistance against wet and dry heat. This is therefore the rate-limiting step in the high pressure sterilization process. To evaluate the impact of a broad pressure, temperature and time domain on the DPA release, Bacillus subtilis spores were pressure treated between 0.1 and 900 MPa at between 30 and 80 °C under isothermal isobaric conditions during dwell time. DPA quantification was assessed using HPLC, and samples were taken both immediately and 2 h after the pressure treatment. To obtain a release kinetic for some pressure-temperature conditions, samples were collected between 1s and 60 min after decompression. A multiresponse kinetic model was then used to derive a model covering all kinetic data. The isorate lines modeled for the DPA release in the chosen pressure-temperature landscape enabled the determination of three distinct zones. (I) For pressures <600 MPa and temperatures >50 °C, a 90% DPA release was achievable in less than 5 min and no difference in the amount of DPA was found immediately 2 h after pressurization. This may indicate irreversible damage to the inner spore membrane or membrane proteins. (II) Above 600 MPa the synergism between pressure and temperature diminished, and the treatment temperature alone dominated DPA release. (III) Pressures <600 MPa and temperatures <50 °C resulted in a retarded release of DPA, with strong increased differences in the amount of DPA released after 2 h, which implies a pressure-induced physiological like germination with cortex degradation, which continues after pressure release. Furthermore, at 600 MPa and 40 °C, a linear relationship was found for the DPA release rate constants ln(k(DPA)) between 1 and 30 min.

(Ultra) High Pressure Homogenization for Continuous High Pressure Sterilization of Pumpable Foods – A Review

Bacterial spores have a strong resistance to both chemical and physical hurdles and create a risk for the food industry, which has been tackled by applying high thermal intensity treatments to sterilize food. These strong thermal treatments lead to a reduction of the organoleptic and nutritional properties of food and alternatives are actively searched for. Innovative hurdles offer an alternative to inactivate bacterial spores. In particular, recent technological developments have enabled a new generation of high pressure homogenizer working at pressures up to 400 MPa and thus, opening new opportunities for high pressure sterilization of foods. In this short review, we summarize the work conducted on (ultra) high pressure homogenization (U)HPH to inactivate endospores in model and food systems. Specific attention is given to process parameters (pressure, inlet, and valve temperatures). This review gathers the current state of the art and underlines the potential of UHPH sterilization of pumpable foods while highlighting the needs for future work.

Shift of pH-Value During Thermal Treatments in Buffer Solutions and Selected Foods

The pH value is one of the most important process parameters during thermal treatments, regardless if the medium is a simple buffer solution or a complex food matrix. When the temperature increases after an initial measurement of the pH at ambient temperature (25°C), a significant pH shift could occur, which could produce incomparable results in different buffer solutions or lead to side reactions during food preservation. Consequently, a measurement cell was constructed to record online the pH-value and temperature up to 130°C. By applying the Nernst equation, it was possible to exclude the temperature-dependent influence of the pH glass electrode on the total pH value. The pH shift was measured over a wide temperature range (ΔT 20–130°C) in the most commonly used buffer solutions and some selected food matrices. The ΔpH of certain buffer solutions, namely TRIS and ACES, showed a significant pH decrease of −2.01 ± 0.08 (ΔT 20–130°C) and −1.27 ± 0.1 (ΔT 20–130°C), respectively, whereas the pH of PBS buffer solution was nearly independent of temperature. The ΔpH decrease recorded in milk (−0.89 ± 0.6, ΔT 20–130°C) as well as commercial and self-made baby food (−0.56 ± 0.05, ΔT 20–130°C) is of special interest for the food industry to get a deeper insight in occurring reactions during thermal preservation processes.

Ultra high pressure homogenization (UHPH) inactivation of Bacillus amyloliquefaciens spores in phosphate buffered saline (PBS) and milk

Ultra high pressure homogenization (UHPH) opens up new areas for dynamic high pressure assisted thermal sterilization of liquids. Bacillus amyloliquefaciens spores are resistant to high isostatic pressure and temperature and were suggested as potential surrogate for high pressure thermal sterilization validation. B. amyloliquefaciens spores suspended in PBS buffer (0.01 M, pH 7.0), low fat milk (1.5%, pH 6.7), and whole milk (3.5%, pH 6.7) at initial concentration of ~106 CFU/mL were subjected to UHPH treatments at 200, 300, and 350 MPa with an inlet temperature at ~80°C. Thermal inactivation kinetics of B. amyloliquefaciens spores in PBS and milk were assessed with thin wall glass capillaries and modeled using first-order and Weibull models. The residence time during UHPH treatments was estimated to determine the contribution of temperature to spore inactivation by UHPH. No sublethal injury was detected after UHPH treatments using sodium chloride as selective component in the nutrient agar medium. The inactivation profiles of spores in PBS buffer and milk were compared and fat provided no clear protective effect for spores against treatments. Treatment at 200 MPa with valve temperatures lower than 125°C caused no reduction of spores. A reduction of 3.5 log10CFU/mL of B. amyloliquefaciens spores was achieved by treatment at 350 MPa with a valve temperature higher than 150°C. The modeled thermal inactivation and observed inactivation during UHPH treatments suggest that temperature could be the main lethal effect driving inactivation.

High-Pressure-Induced Effects on Bacterial Spores, Vegetative Microorganisms, and Enzymes

High pressure (HP) processing is an emerging technology used in the food industry. At present, the process pressure ranges from 350 to 800 MPa in ultra HP homogenizers or isostatic HP units, whereas the majority of isostatic HP application is used for food pasteurization at chilled and ambient temperature. In the last decade HP technology was extended to a broad range of products, and the number of industrial HP systems has steadily increased; at present more than 156 industrial pasteurized products (with an annual production of 300,000 t) are in use worldwide.

Temperature control for high pressure processes up to 1400 MPa

Pressure- assisted sterilisation is an emerging technology. Hydrostatic high pressure can reduce the thermal load of the product and this allows quality retention in food products. To guarantee the safety of the sterilisation process it is necessary to investigate inactivation kinetics especially of bacterial spores. A significant roll during the inactivation of microorganisms under high pressure has the thermodynamic effect of the adiabatic heating. To analyse the individual effect of pressure and temperature on microorganism inactivation an exact temperature control of the sample to reach ideal adiabatic conditions and isothermal dwell times is necessary. Hence a heating/cooling block for a high pressure unit (Stansted Mini-Food-lab; high pressure capillary with 300 μL sample volume) was constructed. Without temperature control the sample would be cooled down during pressure built up, because of the non-adiabatic heating of the steel made vessel. The heating/cooling block allows an ideal adiabatic heat up and cooling of the pressure vessel during compression and decompression. The high pressure unit has a pressure build-up rate up to 250 MPa s-1 and a maximum pressure of 1400 MPa. Sebacate acid was chosen as pressure transmitting medium because it had no phase shift over the investigate pressure and temperature range. To eliminate the temperature difference between sample and vessel during compression and decompression phase, the mathematical model of the adiabatic heating/cooling of water and sebacate acid was implemented into a computational routine, written in Test Point. The calculated temperature is the setpoint of the PID controller for the heating/cooling block. This software allows an online measurement of the pressure and temperature in the vessel and the temperature at the outer wall of the vessel. The accurate temperature control, including the model of the adiabatic heating opens up the possibility to realise an ideal adiabatic heating and cooling as well as isotherm dwell time of the sample under high pressure. This research offers a useful tool to investigate the additive effect of high pressure and thermal treatment on the inactivation of microorganisms.