Inactivation Technologies Research Articles

Pulsed electric field inactivation of microorganisms: from fundamental biophysics to synergistic treatments.

The growth of antibiotic resistant microorganisms and the increasing demand for nonthermal antimicrobial treatment in the food and beverage industry motivates research into alternative inactivation methods. Pulsed electric fields (PEFs) provide an athermal method for inactivating microorganisms by creating nanometer-sized membrane pores in microorganisms, inducing cell death when the PEF duration and intensity are sufficient such that the pores cannot reseal after the PEFs through a process referred to as irreversible electroporation. While PEF inactivation has been studied for several decades, recent studies have focused on extending the technique to various liquids in the food industry and optimizing microorganism inactivation while minimizing adverse effects to the treated sample. This minireview will assess the biophysical mechanisms and theory of PEF-induced cellular interactions and summarize recent advances in applying this technology for microorganism inactivation alone and synergistically in combination with other technologies, including temperature, pressure, natural ingredients, and pharmaceuticals.

Rapid determination of the electroporation threshold for bacteria inactivation using a lab-on-a-chip platform

Electroporation based locally enhanced electric field treatment (LEEFT) is an emerging bacteria inactivation technology for drinking water disinfection. Nevertheless, the lethal electroporation threshold (LET) for bacteria has not been studied, partly due to the tedious work required by traditional experimental methods. Here, a lab-on-a-chip device composed of platinum electrodes deposited on a glass substrate is developed for rapid determination of the LET. When voltage pulses are applied, an electric field with a linear strength gradient is generated on a channel between the electrodes. Bacterial cells exposed to the electric field stronger than the LET are inactivated, while others remain intact. After a cell staining process to differentiate dead and live bacterial cells, the LETs are obtained by analyzing the fluorescence microscopy images. Staphylococcus epidermidis has been utilized as a model bacterium in this study. The LETs range from 10 kV/cm to 35 kV/cm under different pulsed electric field conditions, decreasing with the increase of pulse width, effective treatment time, and pulsed electric field frequency. The effects of medium properties on the LET were also investigated. This lab-on-a-chip device and the experimental approach can also be used to determine the LETs for other microorganisms found in drinking water.

The impact of power ultrasound application on physicochemical, antioxidant, and microbiological properties of fresh orange and celery juice blend

The aim of this work was to study the effect of power ultrasound (PUS) application on physicochemical, antioxidant, and microbiological properties of a juice blend from oranges and celery stalks. Juice samples were sonicated in an ultrasonic bath (20 kHz, 5.57 ± 0.87 kW/m3, 20 ± 5 °C) at different sonication times (0, 15, 30, 45, 60, 75, and 90 min). Samples were stored in the dark for 24 h at 4 °C and subsequently analyzed. Significant (p 0.60; p < 0.001) with an increase in bioactive compound content and antioxidant activity. Also, the 53% increase in cloud value (1.70 ± 0.03) points to an increased stability of the PUS-treated juice blend. Therefore, ultrasonic treatment could represent a viable non-thermal technology for microbial inactivation in fresh orange and celery juice blend while increasing its stability and preserving its bioactive properties and quality attributes.

Haemostatic responsiveness and release of biological response modifiers following cryopreservation of platelets treated with amotosalen and ultraviolet A light.

Due to the risk of replication of contaminating pathogens, platelets have a limited storage time of 5 days, which can be prolonged to 7 days by the use of pathogen inactivation technologies. Cryopreservation (CP) may be an alternative to permit longer storage periods and increased availability. However, the preparation of platelets can result in secretion of biological response modifiers (BRM), which can cause adverse transfusion reactions in the recipient. We investigated the impact of CP on platelet function and release of BRM in untreated (conventional) and pathogen-inactivated (PI) platelet concentrates. Twelve buffy coat-derived platelet units were treated with amotosalen and ultraviolet A light to inactivate pathogens. Twelve untreated units were used as controls. The 24 units were cryopreserved and in vitro variables were analysed before and after CP. The in vitro variables investigated included platelet surface receptors and activation markers by flow cytometry, and coagulation time by viscoelastography. A panel of BRM, including cytokines, was investigated. CP of both conventional and PI platelets resulted in a significant increase of BRM with similar increases of most of the BRM after CP of conventional and PI platelet concentrates. The increase in some of the BRM correlated significantly with shortened coagulation time, increased P-selectin expression, reduced mitochondrial transmembrane potential, and reduced capacity to respond to stimulation with ADP and collagen. Cryopreservation of both conventional and PI platelets results in secretion of BRM. The increase in some of the BRM correlated with changes in platelet function variables and suggests that BRM release is affected, in part, in a similar way by CP as are changes in platelet function variables. PI with amotosalen and ultraviolet A light in combination with CP did not affect the release of immunomodulatory factors more than CP alone did.

Proceedings of the Food and Drug Administration public workshop on pathogen reduction technologies for blood safety 2018 (Commentary, p. 3026).

On November 29, 2018, experts in the field of infectious diseases, pathogen reduction technologies (PRTs) and other participants from blood centers, academia, and industry gathered at the Food and Drug Administration (FDA) White Oak Campus in Silver Spring, Maryland, for a 2‐day public workshop entitled “Pathogen Reduction Technologies for Blood Safety.” The workshop opened with welcome remarks from Dr. Nicole Verdun, Director, Office of Blood Research and Review (OBRR), Center for Biologics Evaluation and Research (CBER), FDA, followed by introductory remarks from Dr. Peter Marks, Director, CBER, FDA. The first day of the workshop focused on blood‐borne infectious agents and their impact on blood safety, experiences of the American Red Cross, and other blood establishments in implementing FDA‐approved pathogen inactivation (PI) technology for plasma and platelets (PLTs) in the United States and novel PRTs under consideration for whole blood (WB) and red blood cells (RBCs). The second day opened with welcome remarks from Dr. Chintamani Atreya, Associate Director for Research, OBRR, CBER, FDA. The focus was on emerging innovations relevant to PRTs and potential alternatives to PRTs. The workshop concluded with remarks on insights for future research and development in this area for blood and blood product safety from infectious agents. A brief introduction of each session by the session moderator followed by a summary of the speaker presentation as submitted by the moderator and speaker are reported here.

Inactivation of yellow fever virus in plasma after treatment with methylene blue and visible light and in platelet concentrates following treatment with ultraviolet C light.

Yellow fever virus (YFV) is endemic to tropical and subtropical areas in South America and Africa, and is currently a major public health threat in Brazil. Transfusion transmission of the yellow fever vaccine virus has been demonstrated, which is indicative of the potential for viral transfusion transmission. An approach to manage the potential YFV transfusion transmission risk is the use of pathogen inactivation (PI) technology systems, such as THERAFLEX MB-Plasma and THERAFLEX UV-Platelets (Macopharma). We aimed to investigate the efficacy of these PI technology systems to inactivate YFV in plasma or platelet concentrates (PCs). YFV spiked plasma units were treated using THERAFLEX MB-Plasma system (visible light doses: 20, 40, 60, and 120 [standard] J/cm2 ) in the presence of methylene blue (approx. 0.8 μmol/L) and spiked PCs were treated using THERAFLEX UV-Platelets system (ultraviolet C doses: 0.05, 0.10, 0.15, and 0.20 [standard] J/cm2 ). Samples were taken before the first and after each illumination dose and tested for residual virus using a modified plaque assay. YFV infectivity was reduced by an average of 4.77 log or greater in plasma treated with the THERAFLEX MB-Plasma system and by 4.8 log or greater in PCs treated with THERAFLEX UV-Platelets system. Our study suggests the THERAFLEX MB-Plasma and the THERAFLEX UV-Platelets systems can efficiently inactivate YFV in plasma or PCs to a similar degree as that for other arboviruses. Given the reduction levels observed in this study, these PI technology systems could be an effective option for managing YFV transfusion-transmission risk in plasma and PCs.

Plasmon-Enhanced Pan-Microbial Pathogen Inactivation in the Cavitation Regime: Selectivity Without Targeting

Pan-microbial inactivation technologies that do not require high temperatures, reactive chemical compounds, or UV radiation could address gaps in current infection control strategies and provide efficient sterilization of biologics in the biotechnological industry. Here, we demonstrate that femtosecond (fs) laser irradiation of resonant gold nanoparticles (NPs) under conditions that allow for E-field mediated cavitation and shockwave generation achieve an efficient plasmon-enhanced photonic microbial pathogen inactivation. We demonstrate that this NP-enhanced, physical inactivation approach is effective against a diverse group of pathogens, including both enveloped and non-enveloped viruses, and a variety of bacteria and mycoplasma. Photonic inactivation is wavelength-dependent and in the absence of plasmonic enhancement from NPs, negligible levels of microbial inactivation are observed in the near-infrared (NIR) at 800 nm. This changes upon addition of resonant plasmonic NPs, which provide a strong enhancement of inactivation of viral and bacterial contaminants. Importantly, the plasmon-enhanced 800 nm femtosecond (fs)-pulse induced inactivation was selective to pathogens. No measurable damage was observed for antibodies included as representative biologics under identical conditions.

Effect of airborne ultrasonic technology on microbial inactivation and quality of dried food ingredients

Conventional methods for decontamination of dried foods have various limitations including undesired effects on both food quality and nutritional parameters. Airborne acoustic technology is a potential technology for microbial inactivation which has limited impacts on food quality. The objective of this study was to investigate the potential of airborne acoustic technology for the decontamination of black pepper grains (Piper nigrum L.) and tapioca starch (Manihot esculenta) inoculated with Bacillus subtilis vegetative cells and spores. Black pepper and tapioca starch samples were subjected to airborne acoustic waves at 170 W for treatment times of 0 (control) to 120 min at room temperature. Significant reductions of 2.19 log CFU/g and 2.01 log CFU/g were achieved after 30 min of treatment for black pepper and tapioca starch respectively (p < 0.05). No significant reductions were observed in spore contaminated samples (p > 0.05). Total phenolic content of black pepper and rheological properties of tapioca starch were not significantly influenced by airborne acoustic treatment at treatment times required to achieve 2 log reductions (p > 0.05). This study demonstrated that airborne acoustic technology can be employed for microbial inactivation of dried food ingredients with minimal impacts on food quality.

Development of a microchip based on LIGA technology for bacterial inactivation

We designed and fabricated a new micro-needle-column discharge chip by Lithographie, Galvanoformung and Abformung technology to meet sterilization requirements. The dimensions of the chip are 11.6 mm × 11.7 mm × 3 mm. The micro-discharge structure is composed of a needle electrode and two cylinder electrodes. We employed a needle with a tip diameter of 20 [Formula: see text]m and a thickness of 20 [Formula: see text]m as the discharge cathode. We used a cylinder with a thickness of 1 mm and diameter of 0.4 mm as the anode. We carried out the gas discharge experiments in ambient air and at room temperature, without external air flow. In the experiments, a corona and glow discharge with applied voltages of −2.1 kV and −3.8 kV were realized. We carried out the sterilization experiments for six different discharge times (15, 30, 60, 90, 120 and 180 s) with an applied voltage of −3.8 kV. The inactivation efficiency of Escherichia coli and Staphylococcus aureus reached 100% for sterilization times of 120 s and 180 s. This indicates that the micro-needle-column discharge chip may be suitable as a portable sterilizer.

Influence of plasma characteristics on the efficacy of Cold Atmospheric Plasma (CAP) for inactivation of Listeria monocytogenes and Salmonella Typhimurium biofilms

Abstract The biofilm mode of growth protects bacterial cells from applied disinfection methods for abiotic (food) contact surfaces. Therefore, new inactivation technologies such as Cold Atmospheric Plasma (CAP) should be considered. However, the influence of different plasma characteristics on the CAP efficacy for biofilm inactivation requires further study. In this research, the influence of (i) the applied plasma configuration (Dielectric Barrier Discharge (DBD) and Surface Barrier Discharge (SBD)), (ii) the oxygen level of the gas flow (He + 0.0/0.5/1.0 (v/v) % O2), and (iii) the plasma intensity (13.88, 17.88, and 21.88 V input voltage) on the CAP efficacy for inactivation of L. monocytogenes and S. Typhimurium biofilms was investigated. Depending on the applied plasma characteristics, log10-reductions up to approximately 3.5 log(CFU/cm2) were obtained. Nevertheless, it could be concluded that the highest log-reductions were in general obtained while using the DBD electrode, 0.0 (v/v) % O2, and an input voltage of 21.88 V. Industrial relevance This study demonstrated the potential application of CAP for inactivation of pathogenic biofilms developed on abiotic (food) contact surfaces. The effect of different plasma characteristics on the CAP inactivation efficacy was investigated and determined optimal conditions resulted in promising reductions of the biofilm-associated cells. By incorporating this novel technology in a complete cleaning and disinfection process, the risk of (cross) contamination of food products might extensively be reduced.

Advanced oxidation technology for the development of a next-generation inactivated West Nile virus vaccine

West Nile virus (WNV) is the most frequent mosquito-borne disease reported in the continental United States and although an effective veterinary vaccine exists for horses, there is still no commercial vaccine approved for human use. We have previously tested a 3% hydrogen peroxide (H2O2)-based WNV inactivation approach termed, HydroVax, in Phase I clinical trials and the vaccine was found to be safe and modestly immunogenic. Here, we describe an advanced, next-generation oxidation approach (HydroVax-II) for the development of inactivated vaccines that utilizes reduced concentrations of H2O2 in combination with copper (cupric ions, Cu2+) complexed with the antiviral compound, methisazone (MZ). Further enhancement of this oxidative approach included the addition of a low percentage of formaldehyde, a cross-linking reagent with a different mechanism of action that, together with H2O2/Cu/MZ, provides a robust two-pronged approach to virus inactivation. Together, this new approach results in rapid virus inactivation while greatly improving the maintenance of WNV-specific neutralizing epitopes mapped across the three structural domains of the WNV envelope protein. In combination with more refined manufacturing techniques, this inactivation technology resulted in vaccine-mediated WNV-specific neutralizing antibody responses that were 130-fold higher than that observed using the first generation, H2O2-only vaccine approach and provided 100% protection against lethal WNV infection. This new approach to vaccine development represents an important area for future investigation with the potential not only for improving vaccines against WNV, but other clinically relevant viruses as well.

Irreversible oxidations of platelet proteins after riboflavin-UVB pathogen inactivation.

Pathogen inactivation technologies are known to alter in vitro phenotype and functional properties of platelets. Because pathogen inactivation generates reactive oxygen species, oxidative stress is considered as one of the plausible cause at the origin of the platelet storage lesion acceleration after treatment. To date proteomics has been used to document the protein variations to picture out the impact. Here, platelet concentrates were prepared from buffy-coats in Intersol additive solution, leukoreduced and pathogen inactivated using a riboflavin/UVB treatment. At day 2 of storage the platelet proteomes of control (untreated) and treated platelet concentrates were investigated against the site specific oxidation by liquid chromatography coupled to tandem mass spectrometry in a shotgun experiment. The shotgun approach detected 9350 peptides (and 2534 proteins) of which 1714 were oxidized. Eighteen peptides were found exclusively oxidized in treated platelets whereas 3 peptides were only found oxidized in control. The present data evidenced an interference with several proteins involved in platelet aggregation and platelet shape change (such as talin and vinculin).

In vitro evaluation of pathogen inactivated platelet quality: An 8 year experience of routine use in Galicia, Spain

BackgroundPlatelet concentrates (PCs) treated by the pathogen inactivation technology (PI) using amotosalen and UVA illumination (PI-PCs) can be manufactured in additive solutions (PAS-III and PAS-IIIM) or in 100% Plasma. Quality control (QC) is an integral part of the production. We capitalized on our ongoing QC program to capture 8 years-worth of data on parameters related to the quality of 116,214 PI-PCs produced under different manufacturing methods. Materials and methodsSelected in vitro parameters of metabolism, activation, and storage were analyzed for the different manufacturing periods to compare PI-PCs versus conventional PCs (C-PCs) resuspended in different PAS. Results and discussionAll BC-PCs met quality standards for pH and dose and residual leucocytes. As expected, storage time correlated with increased lactate, LDH, Annexin V, CD62, sCD40 L levels and decreased glucose and pH. With PAS-IIIM, higher levels of glucose were observed toward the end of shelf life (p < 0.0001) with lower platelet activation markers Annexin V (p = 0.038) and CD62 (p = 0.0006). Following PI implementation, a low expire rate of <0.5% was observed. While a 2.3% mean increase in the production of PCs occurred from 2011 to 2015, the distribution of red blood cell concentrates dropped by 4.4%. A mean incidence of 0.14% for transfusion-related adverse reaction was observed while PI-PCs were distributed, similar to the one observed with C-PCs. Overall, PI-PCs prepared in additive solutions consistently met quality standards. Those prepared in PAS-IIIM appeared to have better retention of in vitro characteristics compared to PAS-III though all demonstrated functionality and clinical effectiveness.

Pectin methylesterase inactivation by High Pressure Carbon Dioxide (HPCD)

In the present work the effect of High Pressure Carbon Dioxide (HPCD) on the activity and structure of a commercial pectin methylesterase (PME) was studied and the results were compared to the conventional thermal inactivation technology. The use of supercritical CO2 (pressure 6–18 MPa, temperature 40–55 °C and time up to 75 min) increased dramatically the PME inactivation rate. The experimental data fitted the first order model and the inactivation kinetic study of PME was completed with the calculations of the activation energy and volume of activation.HPCD treatment induced important changes in the tertiary structure of the enzyme, as revealed the intrinsic fluorescence, KI quenching and ANS binding analyses. They showed that HPCD treatment rearranged the structure of the enzyme. These changes induced by HPCD were significantly different from those induced by mild thermal treatment. Finally, the structural changes detected correlated with the enzyme activity losses observed.

Visible-light-driven photocatalytic inactivation of bacteriophage f2 by Cu-TiO2 nanofibers in the presence of humic acid

Pathogenic viruses in drinking water are great threats to public health. Visible-light-driven photocatalysis is a promising technology for virus inactivation. However, the existing photocatalytic antiviral research studies have mostly been carried out in single-component systems, neglecting the effect of natural organic matter, which exists widely in actual water bodies. In this paper, electrospun Cu-TiO2 nanofibers were prepared as photocatalysts, and their photocatalytic antiviral performance in the presence of humic acid (HA) was comprehensively studied for the first time. The properties of the reaction mixture were measured during the reaction. In addition, the safety, reliability and stability of photocatalytic disinfection in the mixed system were evaluated. The results showed that the virus removal efficiency decreased with the increase of the HA concentration. The type of reaction solution, such as PBS buffer solution or water, did not affect the removal efficiency noticeably. Under acidic conditions, the electrostatic forces between photocatalysts and viruses were strengthened, leading to higher virus removal efficiency. As the reaction time went on, the pH value in the solution increased first and then tended to be stable, the conductivity remained stable, and the dissolved oxygen increased first and then decreased. The safety test showed that the concentration of Cu ions released into the solution was lower than specified by the international standards. No photoreactivation was observed, and the addition of HA significantly reduced the reutilization efficiency of the photocatalysts.

Platelet transfusion associated risks and improvement of platelet transfusion safety with Amotosalen/UVA pathogen inactivation technology

Despite the introduction of multiple measures to minimize the risk of transfusion-transmitted infections, there is still a residual risk of platelet transfusions, especially but not only for bacterial sepsis. Recently, the importance of a reliable hemovigilance system has been underlined and an up to 10-fold difference in the reporting of bacterial transmissions between active and passive reporting has been demonstrated. Another reason for a misjudging of the blood safety may derive from the fact that some pathogens cause obvious damage to critically ill and/or immunocompromised patients only and asymptomatic infections in immuno-competent recipients and thus are not being reported. Pathogen inactivation for platelets, a proactive approach not only broadly inactivating pathogens, but also white blood cells, could minimize the risk of transfusion transmitted infections and graft versus host disease due to residual leukocytes. Only the INTERCEPT technology has received approvals from the regulatory agencies in France, Germany and Switzerland, United States and Canada. That technology utilizes photoactive methods to modify nucleic acids. Long-term routine clinical experience, also with children and neonates, shows the safety and efficacy of INTERCEPT platelet transfusions. The national hemovigilance data of Switzerland, France and Belgium as well as single-center routine use studies show an improved clinical outcome in the acute as well as in the prophylactic setting with a significant decrease in septic and other non-hemolytic transfusion reactions, as well as prevention of graft-versus-host disease.

Efficacy of UV-TiO2 photocatalysis technology for inactivation of Escherichia coli K12 on the surface of blueberries and a model agar matrix and the influence of surface characteristics

Surface disinfection of fresh blueberries is an important food safety challenge due to the delicate texture and short shelf life of these small fruits. A newly designed water-assisted photocatalytic reactor was developed for disinfection of fruits with a delicate texture and complex surface characteristics. Efficacy of UV-TiO2 photocatalysis was evaluated in comparison with UV alone for inactivation of Escherichia coli K12 (as a surrogate for Escherichia coli O157:H7) inoculated onto the surface of the blueberry skin, calyx, and an experimentally prepared agar matrix that was used as a model matrix. Influence of surface characteristics such as surface hydrophobicity and surface free energy on bacterial adhesion were also investigated. The initial bacterial population on all surfaces was approximately 7.0 log CFU/g. UV-TiO2 photocatalysis (4.5 mW/cm2) for 30 s achieved comparatively higher bacterial reductions of 5.3 log and 4.6 log CFU/g on blueberry skin and agar matrix surfaces, respectively, than 4.5 log and 3.4 log CFU/g reductions for UV alone (6.0 mW/cm2). Total phenolic and total anthocyanin contents of fruits were significantly increased after both UV-TiO2 and UV treatments, compared with water washed control fruits. UV-TiO2 photocatalysis technology is a non-chemical and residue-free method with reduced water usage for surface disinfection of fresh blueberries.

Pathogen inactivation/reduction technologies for platelet transfusion: Where do we stand?

Pathogen inactivation or reduction technologies for platelet components have been proposed to secure the microbial safety of this component, and particularly the microbial contamination. Transfusion-transmitted bacterial infections were substantially reduced by additive steps applied at different levels of the transfusion chain, but still killed one recipient each year or every other year in a country like France. Besides, pathogen inactivation and reduction stand for eliminating most viral and protozoa infections persisting in the separated blood component. In addition, because those processes attack nucleic acids, they also aim at substantially alleviating the risk of Transfusion Associates Graft versus Host Disease, as they attack lymphocytes still comprises within the component. Meanwhile, pathogen inactivation or reduction may inflict some damages to the platelet components, that are shown to be additive to the “natural” storage lesions linked to ageing. While it is in general assumed that such processes do not expose transfused patients to an over-risk of bleeding, and are safe, this does not mean that there are no detrimental consequences in the patients, even if not ascribed to as serious. Two such effects are now discussed or debated: the first one is the possible increase in the demand of platelet component, and the other one could be a possible risk of alloimmunisation especially when treated platelets are aged (over 5 days). Three processes have been made available by the industry, that differ in their chemical and physical (ultraviolet light illumination) characteristics. Two processes are largely used (one nationwide in two European countries) and the third one is still under clinical evaluation. This short review endeavored to critically present the main features of the processes and of their implementation.

Pulsed electric field (PEF) technology for microbial inactivation in low-alcohol red wine.

The decontamination of spoilage-related microbes in low-alcohol red wine was performed using a serial multiple electrode pulsed electric field (PEF) treatment system. The system consisted of seven electrodes connected in series, and it has been designed to produce square-wave high-voltage pulses of 1μs duration at various electric field strengths and frequencies for decontamination. The initial counts of aerobic bacteria, yeast and lactic acid bacteria (spoilage-associated microbes) in the wine were 5.56, 5.61 and 5.22logCFU/mL, respectively. The pattern of decontamination of the spoilage microorganisms followed first-order kinetics and the decontamination effect increased as the field strength and frequency increases. DHz and DPEF values were inversely related to the electric field strength of the PEF treatment. The yeast exhibited relatively low DPEF-value than the aerobic and lactic acid bacteria. The lowest ZPEF-value was observed for the lactic acid bacteria (24.6kV/cm) among the spoilage microbes.

High Intensity Ultrasound for Salmonella Enteritidis Inactivation in Culture and Liquid Whole Eggs.

High intensity ultrasound (HIU) continues to be researched as a nonthermal inactivation technology of appeal to food manufacturers. The advantages of HIU include maintenance of product quality, freshness, product homogenization, along with simultaneous inactivation of pathogens. Besides, it is simple, relatively inexpensive, and easily adaptable to most processing environments. This study investigated the effect of HIU for Salmonella Enteritidis inactivation in culture and liquid whole eggs (LWEs) to decrease egg-associated outbreaks. Overnight S. Enteritidis cultures and spiked LWE (both at 8 log CFU/mL) were treated with 20-kHz HIU for 0, 1, 5, 10, and 30min (n=6) in a temperature-controlled system, not to exceed 20 °C, and replicated thrice. At each time point, samples were enumerated on XLT4 agar and morphologically analyzed using scanning electron microscopy, with measurements of color and rheological properties. Our results revealed significant reduction of healthy S. Enteritidis cells up to 3.6log CFU/mL and 2.3log CFU/25mL after HIU treatment of merely 10min of overnight culture and 30min in LWE, respectively (P < 0.05). After 5 and 10-min HIU treatment, significant reduction of 1.4-log CFU/25mL healthy S. Enteritidis in LWE was obtained (P<0.05). Even at 1-min exposure time, HIU showed significant 1.9 log CFU/mL reduction of cultures (P<0.05); however, no log-reduction was observed in LWE after 1 min. Scanning electron micrographs showed increased cell structural damage using longer HIU exposure. For product color changes, lower redness and yellowness of LWE were observed visually and instrumentally after 5-min HIU treatment (P<0.05). The rheological properties of LWE measured at 0 to 200s-1 shear rate, showed that shear stress of HIU-treated LWEs decreased after 5-min HIU exposure, but increased after 30-min treatment. This study demonstrated that HIU shows promise for rapid Salmonella control in LWE and other liquid foods, as an alternative inactivation method for use in hurdle approaches.