Inactivation of foodborne pathogens by the synergistic combinations of food processing technologies and food‐grade compounds

There is a need to develop food processing technologies with enhanced antimicrobial capacity against foodborne pathogens. While considering the challenges of adequate inactivation of pathogenic microorganisms in different food matrices, the emerging technologies are also expected to be sustainable and have a minimum impact on food quality and nutrients. Synergistic combinations of food processing technologies and food‐grade compounds have a great potential to address these needs. During these combined treatments, food processes directly or indirectly interact with added chemicals, intensifying the overall antimicrobial effect. This review provides an overview of the combinations of different thermal or nonthermal processes with a variety of food‐grade compounds that show synergistic antimicrobial effect against pathogenic microorganisms in foods and model systems. Further, we summarize the underlying mechanisms for representative combined treatments that are responsible for the enhanced microbial inactivation. Finally, regulatory issues and challenges for further development and technical transfer of these new approaches at the industrial level are also discussed.     

Combination of Pulsed Electric Fields with Other Preservation Techniques

Pulsed electric field (PEF) is a promising nonthermal treatment for liquid foods with antimicrobial activity at ambient temperature. According to the hurdle concept, the combination of PEF with other preservation methods may enhance cell death. The joint effect of PEF with the addition of antimicrobial compounds of natural origin, such as nisin, has received special attention, although other emerging nonthermal techniques, such as the use of high-pressure carbon dioxide, have also been considered. Moreover, the bactericidal action resulting from the simultaneous application of PEF and conventional thermal treatments suggests the possibility of lowering the intensity of heating while maintaining microbial acceptance. This review analyses the literature on antimicrobial strategies that combine PEF with nonthermal and traditional preservation methods. The variables affecting cell death produced by those combined treatments and their mechanisms of synergy are considered. Commercialization of PEF in combination with other technologies may be possible due to its improved antimicrobial effect.     

Nonthermal Food Processing Alternatives and Their Effects on Taste and Flavor Compounds of Beverages

Food drinks are normally processed to increase their shelf-life and facilitate distribution before consumption. Thermal pasteurization is quite efficient in preventing microbial spoilage of many types of beverages, but the applied heat may also cause undesirable biochemical and nutritious changes that may affect sensory attributes of the final product. Alternative methods of pasteurization that do not include direct heat have been investigated in order to obtain products safe for consumption, but with sensory attributes maintained as unchanged as possible. Food scientists interested in nonthermal food preservation technologies have claimed that such methods of preserving foods are equally efficient in microbial inactivation as compared with conventional thermal means of food processing. Researchers in the nonthermal food preservation area also affirm that alternative preservation technologies will not affect, as much as thermal processes, nutritional and sensory attributes of processed foods. This article reviews research in nonthermal food preservation, focusing on effects of processing of food drinks such as fruit juices and dairy products. Analytical techniques used to identify volatile flavor-aroma compounds will be reviewed and comparative effects for both thermal and nonthermal preservation technologies will be discussed.     

Nonthermal preservation of foods using combined processing techniques

In the last 2 decades, consumer demand for fresher, higher quality, and safer food has promoted research on nonthermal methods of food preservation for the inactivation of microorganisms and enzymes as an alternative to thermal processes. However, the high resistance of certain enzymes and microorganisms to nonthermal processes, especially bacterial spores, limit their application. To expand the use of nonthermal processes in the food industry, combinations of these technologies with traditional or emerging food preservation techniques are being studied. The use of nonthermal processes in combination with other preservation technologies presents a number of potential benefits to food preservation. The purpose of this article is to review some successful combinations of different nonthermal technologies, such as high hydrostatic pressure, ultrasound, pulsed electric fields, and irradiation, with traditional or emerging food preservation technologies.     

Quality-Related Enzymes in Plant-Based Products: Effects of Novel Food-Processing Technologies Part 3: Ultrasonic Processing

High-power ultrasound is a versatile technology which can potentially be used in many food processing applications including food preservation. This is part 2 of a series of review articles dealing with the effectiveness of nonthermal food processing technologies in food preservation focusing on their effect on enzymes. Typically, ultrasound treatment alone does not efficiently cause microbial or enzyme inactivation sufficient for food preservation. However, combined with mild heat with or without elevated pressure (P ≤ 500 kPa), ultrasound can effectively inactivate enzymes and microorganisms. Synergistic effects between ultrasound and mild heat have been reported for the inactivation of both enzymes and microorganisms. The application of ultrasound has been shown to enhance the rate of inactivation of quality degrading enzymes including pectin methylesterase (PME), polygalacturonase (PG), peroxidase (POD), polyphenol oxidase (PPO), and lipoxygenase (LOX) at mild temperature by up to 400 times. Moreover, ultrasound enables the inactivation of relatively heat-resistant enzymes such as tomato PG1 and thermostable orange PME at mild temperature conditions. The extent to which ultrasound enhances the inactivation rate depends on the type of enzyme, the medium in which the enzyme is suspended, and the processing condition including frequency, ultrasonic intensity, temperature, and pressure. The physical and chemical effects of cavitation are considered to be responsible for the ultrasound-induced inactivation of enzymes, although the dominant mechanism depends on the structure of the enzyme.     

High pressure processing combined with selected hurdles: Enhancement in the inactivation of vegetative microorganisms

High pressure processing (HPP) as a nonthermal processing (NTP) technology can ensure microbial safety to some extent without compromising food quality. However, for vegetative microorganisms, the existence of pressure-resistant subpopulations, the revival of sublethal injury (SLI) state cells, and the resuscitation of viable but nonculturable (VBNC) state cells may constitute potential food safety risks and pose challenges for the further development of HPP application. HPP combined with selected hurdles, such as moderately elevated or low temperature, low pH, natural antimicrobials (bacteriocin, lactate, reuterin, endolysin, lactoferrin, lactoperoxidase system, chitosan, essential oils), or other NTP (CO₂, UV-TiO₂ photocatalysis, ultrasound, pulsed electric field, ultrafiltration), have been highlighted as feasible alternatives to enhance microbial inactivation (synergistic or additive effect). These combinations can effectively eliminate the pressure-resistant subpopulation, reduce the population of SLI or VBNC state cells and inhibit their revival or resuscitation. This review provides an updated overview of the microbial inactivation by the combination of HPP and selected hurdles and restructures the possible inactivation mechanisms.     

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.     

Nonthermal Plasma–Liquid Interactions in Food Processing: A Review

Nonthermal processing methods are often preferred over conventional food processing methods to ensure nutritional quality. Nonthermal plasma (NTP) is a new field of nonthermal processing technology and seeing increased interest for application in food preservation. In food applications of NTP, liquid interactions are the most prevalent. The NTP reactivity and product storability are altered during this interaction. The water activated by NTP (plasma‐activated water [PAW]) has gained considerable attention during recent years as a potential disinfectant in fruits and vegetable washing. However, detailed understanding of the interactions of NTP reactive species with food nutritional components in the presence of water and their stability in food is required to be explored to establish the potential of this emerging technology. Hence, the main objective of this review is to give a complete overview of existing NTP–liquid interactions. Further, their microbial inactivation mechanisms and the effects on food quality are discussed in detail. Most of the research findings have suggested the successful application of NTP and PAW for microbial inactivation and food preservation. Still, there are some research gaps identified and a complete analysis of the stability of plasma reactive species in food is still missing. By addressing these issues, along with the available research output in this field, it is possible that NTP can be successfully used as a food decontamination method in the near future.     

Principles and recent applications of novel non-thermal processing technologies for the fish industry—a review

Thermal treatment is a traditional method for food processing, which can kill microorganisms but also lead to physicochemical and sensory quality damage, especially to temperature-sensitive foods. Nowadays consumers’ increasing interest in microbial safety products with premium appearance, flavor, great nutritional value and extended shelf-life has promoted the development of emerging non-thermal food processing technologies as alternative or substitution to traditional thermal methods. Fish is an important and world-favored food but has a short shelf-life due to its extremely perishable characteristic, and the microbial spoilage and oxidative process happen rapidly just from the moment of capture, making it dependent heavily on post-harvest preservation. The applications of novel non-thermal food processing technologies, including high pressure processing (HPP), ultrasound (US), pulsed electric fields (PEF), pulsed light (PL), cold plasma (CP) and ozone can extend the shelf-life by microbial inactivation and also keep good sensory quality attributes of fish, which is of high interest for the fish industry. This review presents the principles, developments of emerging non-thermal food processing technologies, and also their applications in fish industry, with the main focus on microbial inactivation and sensory quality. The promising results showed great potential to keep microbial safety while maintaining organoleptic attributes of fish products. What’s more, the strengths and weaknesses of these technologies are also discussed. The combination of different food processing technologies or with advanced packaging methods can improve antimicrobial efficacy while not significantly affect other quality properties under optimized treatment.     

Nonthermal Processes for Shelf‐Life Extension of Seafoods: A Revisit

For the past two decades, consumer demand for minimally processed seafoods with good sensory acceptability and nutritive properties has been increasing. Nonthermal food processing and preservation technologies have drawn the attention of food scientists and manufacturers because nutritional and sensory properties of such treated foods are minimally affected. More importantly, shelf‐life is extended as nonthermal treatments are capable of inhibiting or killing both spoilage and pathogenic organisms. They are also considered to be more energy‐efficient and to yield better quality when compared with conventional thermal processes. This review provides insight into the nonthermal processing technologies currently used for shelf‐life extension of seafoods. Both pretreatments such as acidic electrolyte water and ozonification and processing technologies, including high hydrostatic pressurization, ionizing radiation, cold plasma, ultraviolet light, and pulsed electric fields, as well as packaging technology, particularly modified atmosphere packaging, have been implemented to lower the microbial load in seafood. Thus, those technologies may be the ideal approach for the seafood industry, in which prime quality is maintained and safety is assured for consumers.     

Thiol-reactive natural antimicrobials and high pressure treatment synergistically enhance bacterial inactivation

Bacterial inactivation by high pressure (HP) treatment can be strongly enhanced in the presence of antimicrobial compounds, but the mechanism of this synergy is poorly understood. In the present study, screening of thirteen natural antimicrobial compounds (NACs) led to the identification of six NACs that exhibited synergy with HP treatment against eight Gram-negative and two Gram-positive bacteria. The strongest synergy was found with α,β-unsaturated aldehydes (t-cinnamaldehyde, t-2-hexenal, dimethylfumarate), isothiocyanates (allyl isothiocyanate, sulforaphane) and other electrophilic compounds (reuterin). The antibacterial activity of these compounds and their synergistic interaction with HP was linked to thiol reactivity based on (1) their reaction with cysteine in vitro, (2) their reduced minimal inhibitory concentration in an Escherichia coli gshA mutant which is defective in glutathione synthesis, and (3) the loss of synergy with HP in the presence of excess cysteine. These novel insights in the mechanisms leading to synergy will support the development of more effective hurdle technology applications of HP treatment and natural antimicrobials.Industrial relevanceHP treatment is an emerging mild processing technology that offers a better balance between microbiological safety and stability and quality retention compared to traditional heat-based pasteurization methods. The efficacy of microbial inactivation by HP treatment is improved in the presence of natural antimicrobials, but the basis of this synergistic interaction is poorly understood. In the present work, a specific antibacterial mode of action is shown to result in synergy, and this insight will facilitate the development of more effective combined treatments.     

Validation of process technologies for enhancing the safety of low-moisture foods: A review

The outbreaks linked to foodborne illnesses in low-moisture foods are frequently reported due to the occurrence of pathogenic microorganisms such as Salmonella Spp. Bacillus cereus, Clostridium spp., Cronobacter sakazakii, Escherichia coli, and Staphylococcus aureus. The ability of the pathogens to withstand the dry conditions and to develop resistance to heat is regarded as the major concern for the food industry dealing with low-moisture foods. In this regard, the present review is aimed to discuss the importance and the use of novel thermal and nonthermal technologies such as radiofrequency, steam pasteurization, plasma, and gaseous technologies for decontamination of foodborne pathogens in low-moisture foods and their microbial inactivation mechanisms. The review also summarizes the various sources of contamination and the factors influencing the survival and thermal resistance of pathogenic microorganisms in low-moisture foods. The literature survey indicated that the nonthermal techniques such as CO₂, high-pressure processing, and so on, may not offer effective microbial inactivation in low-moisture foods due to their insufficient moisture content. On the other hand, gases can penetrate deep inside the commodities and pores due to their higher diffusion properties and are regarded to have an advantage over thermal and other nonthermal processes. Further research is required to evaluate newer intervention strategies and combination treatments to enhance the microbial inactivation in low-moisture foods without significantly altering their organoleptic and nutritional quality.     

Reaction Kinetics at High Pressure and Temperature: Effects on Milk Flavor Volatiles and on Chemical Compounds with Nutritional and Safety Importance in Several Foods

Consumers demand, in addition to excellent eating quality, high standards of safety and nutrition in ready-to-eat food. This requires a continuous improvement in conventional processing technologies and the development of new alternatives. Prevailing technologies such as thermal processing can cause extensive and undesirable chemical changes in food composition while minimal processing strategies cannot eliminate all microbial pathogens. This review focuses on pressure-assisted thermal processing, a new alternative for shelf-stable foods. Its implementation requires an analysis of reaction kinetics at high pressure and elevated temperature. Acceleration of the inactivation of bacterial spores by the synergistic effect of pressure and temperature is expected to allow processing at lower temperature, shorter process time, or a combination of both. Therefore, thermal degradation of quality is expected to be lower than that of conventional thermal processes. However, few studies have focused on the effect of the conditions required for the inactivation of bacterial spores on the kinetics of chemical reactions degrading food quality, particularly at the high temperatures required for the processing of low-acid foods.     

Alternatives to conventional thermal treatments in fruit-juice processing. Part 1: Techniques and applications

This paper provides an overview of alternatives to conventional thermal treatments and a review of the literature on fruit-juice processing for three key operations in fruit-juice production such as microbial inactivation, enzyme inactivation, and juice yield enhancement, these being radiation treatments (UV light, high-intensity light pulses, γ-irradiation), electrical treatments (pulsed electric fields, radiofrequency electric fields, ohmic heating), microwave heating, ultrasound, high hydrostatic pressure, inert gas treatments (supercritical carbon dioxide, ozonation), and flash-vacuum expansion. The nonthermal technologies discussed in this review have the potential to meet industry and consumer expectations. However, the lack of standardization in operating conditions hampers comparisons among different studies, and consequently ambiguity arises within the literature. For the juice industry to advance, more detailed studies are needed on the scaling-up, process design, and optimization, as well as on the effect of such technologies on juice quality of juices in order to maximize their potential as alternative nonthermal technologies in fruit-juice processing.     

Toward a Philosophy and Theory of Volumetric Nonthermal Processing

Nonthermal processes for food preservation have been under intensive investigation for about the past quarter century, with varying degrees of success. We focus this discussion on two volumetrically acting nonthermal processes, high pressure processing (HPP) and pulsed electric fields (PEF), with emphasis on scientific understanding of each, and the research questions that need to be addressed for each to be more successful in the future. We discuss the character or “philosophy” of food preservation, with a question about the nature of the kill step(s), and the sensing challenges that need to be addressed. For HPP, key questions and needs center around whether its nonthermal effectiveness can be increased by increased pressures or pulsing, the theoretical treatment of rates of reaction as influenced by pressure, the assumption of uniform pressure distribution, and the need for (and difficulties involved in) in‐situ measurement. For PEF, the questions include the rationale for pulsing, difficulties involved in continuous flow treatment chambers, the difference between electroporation theory and experimental observations, and the difficulties involved in in‐situ measurement and monitoring of electric field distribution.     

Cold atmospheric pressure plasma and low energy electron beam as alternative nonthermal decontamination technologies for dry food surfaces: A review

BackgroundDry food products are often highly contaminated, and dry stress-resistant microorganisms, such as certain types of Salmonella and bacterial spores, can be still viable and multiply if the product is incorporated into high moisture food products or rehydrated. Traditional technologies for the decontamination of these products have certain limitations and drawbacks, such as alterations of product quality, environmental impacts, carcinogenic potential and/or lower consumer acceptance. Cold atmospheric pressure plasma (CAPP) and low energy electron beam (LEEB) are two promising innovative technologies for microbial inactivation on dry food surfaces, which have shown potential to solve these certain limitations.Scope and approachThis review critically summarizes recent studies on the decontamination of dry food surfaces by CAPP and LEEB. Furthermore, proposed inactivation mechanisms, product-process interactions, current limitations and upscaling potential, as well as future trends and research needs for both emerging technologies, are discussed.Key findings and conclusionsCAPP and LEEB are nonthermal technologies with a high potential for the gentle decontamination of dry food surfaces. Both technologies have similarities in their inactivation mechanisms. Due to the limited penetration depth of both technologies, product-process interactions can be minimized by maintaining product quality. A first demonstrator with Technology Readiness Level (TRL) 7 for LEEB has already been introduced into the food industry for the decontamination of herbs and spices. Compared with LEEB, CAPP is at the advanced development stage with TRL 5, for which further work is essential to design systems that are scalable to industrial requirements.     

Relationship between optical properties of beverages and microbial inactivation by intense pulsed light

This study determined the inactivation effects of the intense pulsed light (IPL) on Pseudomonas aeruginosa in various liquid foods and elucidated the relationship between microbial reduction and the optical properties of twelve liquid foods. In mineral water and isotonic beverage, 7.0 log reductions of P. aeruginosa were obtained with the IPL treatment at total fluence of 0.97 J/cm². At 12.17–24.35 J/cm², 7.0 log reductions were shown in five liquid samples (apple juice, carbonated beverages and plum juice), while only 0.5- to 2.0-log reductions were shown in the rest samples even after 29.21 J/cm² total fluence of the IPL treatment. High value of transmittance and low value of extinction coefficient have a decisive effect on the microbial inactivation because it allows the intensity of the IPL to be preserved as it penetrates into the sample. The best-fit regression kinetic which can explain the relationship between extinction coefficient and bactericidal effect was an exponential function.Industrial relevanceIntense pulsed light (IPL) is one of the nonthermal processing technologies for ensuring safe foods with satisfactory qualities. Through this study, transparent liquids showed a high microbial reduction level after IPL treatment in a short time. So it can be concluded that IPL has a potential as an excellent alternative or complement to conventional thermal processing of transparent liquids. Also, inactivation kinetic equation deducted from this study can be used to predict the microbial reduction level of specific liquid before IPL treatment by using its extinction coefficient.     

Inactivation Kinetics of Lactobacillus plantarum by High Pressure Carbon Dioxide

Inactivation kinetics of Lactobacillus plantarum by high pressure CO₂ was investigated to understand the mechanism of microbial inactivation. The inactivation rates increased with pressure, temperature and exposure time, and with decreasing pH of media. Microbial inactivation was governed essentially by penetration of CO₂ into cells and its effectiveness could be improved by the enhanced transfer rate. Microbial reduction of 8 log cycles was observed within 120 min under CO₂ pressure of 70 kg/cm² at 30°C. We hypothesized that the cell death resulted from the lowered intracellular pH and damage to the cell membrane due to penetration of CO₂. Pressurized CO₂ treatment is a potential nonthermal technology for food preservation.     

Microbial inactivation of E. coli cells by a combined PEF–HPCD treatment in a continuous flow system

A laboratory scale continuous flow unit was set up and used to study the effect of pulsed electric fields (PEF) pre-treatments on microbial inactivation by high pressure carbon dioxide (HPCD) processing with the aim of investigating the synergistic effect of the combined treatment. McIlvaine buffer solution inoculated with Escherichia coli cells ATCC26 was pre-treated with PEF (25 °C) at different field strength (E = 6–12 kV/cm) and energy input (W_T = 10–40 J/mL) and then processed with HPCD (25 °C) at pressures of 8.0, 14.0 and 20.0 MPa and holding times of 4, 7 and 11 min.Results showed that treating the microbial suspension only with PEF, the inactivation level slightly increased with increasing the field strength and energy input with no significant effect of the pressure applied. The maximum inactivation level obtained was 2.25 Log-cycles at 12 kV/cm and 40 J/mL. When the bacterial cells were treated only with HPCD, the inactivation level was almost independent on the pressure of CO₂, and gradually increased with increasing the holding time up to a maximum value of 2.41 Log-cycles. The combination of PEF and HPCD treatment resulted in a marked increase of the microbial inactivation with increasing the field strength, energy input, holding time and operative pressure. A clear synergistic effect was evident when holding time was longer than 4 min, regardless the intensity of the PEF treatment applied.Industrial relevanceConsumers demand for fresh and natural products forces food manufacturers to investigate milder preservation processes and stimulate the current trend to use hurdle technologies. Pulsed electric field (PEF) and high pressure carbon dioxide (HPCD) are emerging non-thermal technologies which have antimicrobial capabilities when applied alone or in combination with other physicochemical hurdles. The present work demonstrated, for the first time, the feasibility of combined PEF-HCPD process based on the coupling of a PEF pretreatment stage to HPCD treatment in a continuous flow unit. The results support the view that the combined process is able to induce substantial microbial inactivation at mild treatment conditions and at room temperature suggesting the idea that this process could be applied to foods with thermosensitive components.