Virulence of Listeria monocytogenes before and after high hydrostatic pressure treatment

Abstract

High hydrostatic pressure treatment is regarded as a possible alternative process for food preservation. One of the primary considerations for industrial applications is the ability of this technique to destroy pathogenic microorganisms. The inactivation of microorganism populations after high pressure treatment is well described, but their residual pathogenicity is less documented. This study compared the virulence of Listeria monocytogenes scott A before and after high pressure treatments. The pressurized samples were composed of 1 ml of Listeria monocytogenes culture diluted into 9 ml of buffer. Two buffers are used: phosphate buffer (pH 7) and citrate buffer (pH 5.6).

The virulence of cells is estimated by their association and invasion capacities to Hela cells. These characteristics of pathogenity were compared between untreated and pressurized cells (400 MPa, 10min, in citrate buffer) or (600 MPa, 10min, in phosphate buffer). These results showed that, after treatment in citrate buffer, the treated cells lost their association and invasion capacities. However, in a phosphate buffer, treated cells have lose their invasion properties, but retain their association capacities to Hela cells. These results showed that the inactivation of Listeria monocytogenes population involve a loss of their pathogenic properties. However, the conservation of association capacities of some inactivated cells show that these cells seem to continue to express adhesion molecules and, in certain conditions, can reestablish their multiplication properties.     

圆柱形超高静水压水听器设计与测试

(本文介绍了一种应用于20Hz~20kHz频段、最大工作深度5000米的超高静水压水听器,水听器敏感元件采用厚壁压电陶瓷圆管,高强度复合泡沫去耦,通过对敏感元件及水听器结构进行合理设计,保证水听器具有高的耐压强度。设计了一种圆柱形高静压耦合腔,根据耦合腔声学和力学条件,确定了腔体尺寸及厚度,在国内首次实现了最高50MPa压力下的水听器低频灵敏度测试。测试结果表明,水听器常压下接收灵敏度响应平坦,在常压到50MPa压力范围内,水听器低频接收灵敏度最大变化小于2dB,表明该型水听器具有良好的压力稳定性,可在深海资源勘测、深海通讯等方面获得广泛应用。)     

Effect of high hydrostatic pressure on enzyme activity in unpasteurized draft sake

ABSTRACT

Unpasteurized draft sake has a potentially high market value, due to its fresh flavor and fruity taste, compared with conventional thermal-pasteurized sake. However, the shelf life of draft sake is limited. To increase the shelf life of draft sake, it is necessary to suppress flavor and taste deterioration resulting from inactivation of enzymes produced by koji-mold. Draft sake was treated with high hydrostatic pressure (HHP) of 200 to 500?MPa at ?7 to 50°C to analyze the inactivation of α-amylases, glucose-forming enzymes, and acid carboxypeptidases. We found significant inactivation of enzymes produced by koji-mold in draft sake subjected to HHP treatment at both high and low temperature. However, HHP treatment at low temperature effectively inactivated enzymes while retaining the fresh flavor and fruity taste of draft sake.     

Effect of plating and incubation conditions on the detection of lactic acid bacteria after high hydrostatic pressure treatment

ABSTRACT

Lactic acid bacteria (Leuconostoc mesenteroides, Enterococcus faecalis, and Lactobacillus fermentum) were subjected to high hydrostatic pressures (HHPs) of 400 and 600?MPa at 25°C for 10?min in phosphate-buffered saline. Differential plating methods were applied to evaluate HHP-treated cell populations, assuming that healthy and injured cells during plate incubation survived maximum and minimal stress, respectively. The stress was altered by using several selective media in combination with aerobic or anaerobic incubation at 25°C or 30°C. E. faecalis was detectable after 600?MPa treatment while L. mesenteroides and L. fermentum were nondetectable. Specific combinations of incubation conditions were suggested to determine maximum and minimum viable counts of L. mesenteroides and E. faecalis. The difference between the maximum and minimum counts can be used to evaluate HHP-injured population with reduced risks to overestimate healthy and/or underestimate HHP-injured cells.     

The usage of high hydrostatic pressure (HHP) to control food-borne pathogens in hummus

ABSTRACT

With the increasing demand for fresher, higher quality, minimally processed and safer food, there is a strong necessity to develop non-thermal processing techniques. Also for hummus, which is popular all around the world. In this work, the effect of refrigerated storage on the survival of pathogens in hummus treated by high hydrostatic pressure (HHP) (500?MPa/10?min/room temperature) was evaluated. The cocktail of two Salmonella, four Listeria monocytogenes and two Escherichia coli strains was used in this study. All pathogen types were able to survive in hummus during 60 days of refrigerated storage. HHP-treated samples plated on day 0 successfully achieved a?>?5 log cfu/g reduction for all pathogen types. No residual survivors were present after 30 and 60 days in any of the HHP-treated samples. These results demonstrate that HHP may be a useful technique for the inactivation of pathogens and therefore helpful in designing non-thermal HHP conditions for pressurization of hummus.     

Effect of high hydrostatic pressure processing and squeezing pressure on some quality properties of pomegranate juice against thermal treatment

The aim of this study was to investigate the effect of high hydrostatic pressure (HHP) treatment (200, 300, 400?MPa; 5°C, 15°C and 25°C; 5 and 10 min) on some quality properties of pomegranate juice. Juice samples are obtained under industrial conditions at two different squeezing pressure levels (100 and 150?psi – 0.689 and 1.033?MPa, respectively). Results are compared against conventional thermal treatment (85°C/10 min) and raw sample. For all three processing temperature, HHP combinations at 400?MPa for 10 min were sufficient to decrease the microbial load around 4.0 log cycles for both squeeze levels. All HHP treatments showed no significant decrease at antioxidant activity, total phenolic content and monomeric anthocyanin pigment concentrations, while there was a significant decrease (p?≤?.05) in thermal-treated samples. Being the highest sugar alcohol in pomegranate juice, mannitol content must be considered for determining the authenticity, and mannitol content increased with squeezing pressure and thermal treatment.     

应用SRCD和FTIR分析超高压处理对蘑菇多酚氧化酶二级结构的影响

应用同步辐射紫外真空圆二色光谱(SRCD)、傅里叶变换红外光谱(FTIR)和荧光光谱研究了超高压(HHP)处理对蘑菇多酚氧化酶(PPO)二级结构和三级结构的影响。HHP处理使蘑菇PPO的α-螺旋含量明显减少,二级结构发生改变。通过SRCD光谱和FTIR光谱分析得出的未处理或HHP处理蘑菇PPO的二级结构含量均存在一定的差异,这种差异可能是由于测量温度、酶液浓度和分析方法等多种因素造成的。荧光光谱表明,HHP处理后,蘑菇PPO溶液荧光光谱的强度降低,最大发射峰发生了红移,表明HHP处理改变了蘑菇PPO分子的三级结构。     

Effect of high hydrostatic pressure and reducing sodium chloride and phosphate on physicochemical properties of beef gels

ABSTRACT

The effect of high hydrostatic pressure (HHP) treatment (100–200?MPa, 10?min, 20°C) combined with sodium chloride and sodium phosphate on the physicochemical properties of beef gels was investigated. The water content, cooking losses, color, protein composition by SDS-PAGE analysis and texture parameters of beef gels were determined. The beef gels treated with high pressure at 150?MPa showed a synergistic effect in the increased water content and the decreased cooking losses compared with the unpressurized gels. The L*, a* and b* color values of beef gels were slightly decreased under HHP treatment at 100–200?MPa. In the SDS-PAGE analysis, the staining intensity of the α-actinin protein band was decreased in pressurized samples. The cohesiveness, adhesiveness, gel strength and modulus of elasticity were improved after HHP treatment. Application of high pressure treatment (150–200?MPa) before heat treatment would be beneficial for the manufacturing of low salt and/or low phosphate meat products for a healthy diet.     

Effect of ultrasound and high hydrostatic pressure (US/HHP) on the degradation of dextran catalyzed by dextranase

In our current research work, the effect of combination of ultrasonic irradiation and high hydrostatic pressure (US/HHP) on the enzymatic activity and enzymatic hydrolysis kinetic parameters of dextran catalytic by dextranase were investigated. Furthermore, the effects of US/HHP on the structure of dextranase were also discussed with the aid of fluorescence spectroscopy and circular dichroism (CD) spectroscopy. The maximum hydrolysis of dextran was observed under US (40 W at 25 kHz for 15 min) combined with HHP (400 MPa for 25 min), in which the hydrolysis of dextran increased by 163.79% compared with the routine thermal incubation at 50 °C. Results also showed that, V_max and K_M values, as well as, k_cat of dextranase under US/HHP treatment were higher than that under US, HHP and thermal incubation at 50 °C, indicated that, the substrate is converted into the product at an increased rate when compared with the incubation at 50 °C. Compared to the enzymatic reaction under US, HHP, and routine thermal incubation, dextranase enzymatic reaction under US/HHP treatment showed decreases in Ea, ΔG and ΔH, however small increase in ΔS value was observed. In addition, fluorescence and CD spectra reflected that US/HHP treatment had increased the number of tryptophan on dextranase surface with increased α-helix by 19.80% and reduced random coil by 6.94% upon US/HHP-treated dextranase protein compared to the control, which were helpful for the improvement of its activity. These results indicated that, the combination of US and HHP treatments could be an effective method for improving the hydrolysis of dextran in many industrial applications including sugar manufacturing processes.     

Enrichment of free amino acid content and reduction of astringent taste compounds in soybean by high hydrostatic pressure

ABSTRACT

We investigated the effect of high hydrostatic pressure (HHP) on the distribution of free amino acids and isoflavones in soybean immediately after pressure treatment (100–600?MPa, 10–60?min). HHP-treatment at 200 and 300?MPa resulted in high accumulation of free amino acids. Additionally, Gly, Val, and Pro levels increased even after HHP of 400?MPa. The application of HHP-treatment to soybean decreased sources of an astringent taste such as genistein-, daizein-, and glycitein-type molecules under all pressure conditions over 200?MPa. High accumulation of free amino acids with specific pressure–time conditions would be caused by the acceleration of proteolysis and specific amino acid metabolism. We concluded that HHP alone with no subsequent storage enabled the enrichment of specific amino acids such as GABA, Glu, Lys, and Pro in soybean. Soybean with enrichment of these amino acids and improved taste should be an innovative component of functional foods.     

Impacts of sonication and high hydrostatic pressure on the structural and physicochemical properties of quinoa protein isolate dispersions at acidic,neutral and alkaline pHs

Herein, 1 wt% quinoa protein isolate (QPI) was exposed to sonication using a 20 kHz ultrasonicator equipped with a 6 mm horn (14.4 W, 10 mL, up to 15 min) or high hydrostatic pressure (HHP, up to 600 MPa, 15 min) treatments at pH 5, pH 7, and pH 9. The changes to physicochemical properties were probed by SDS-PAGE, FTIR, free sulfhydryl group (SH), surface hydrophobicity (H₀), particle size and solubility. As revealed by SDS-PAGE, substantial amounts of 11S globulin participated in the formations of aggregates via SS bond under HHP, particularly at pH 7 and pH 9. However, protein profiles of QPI were not significantly affected by the sonication. Free SH groups and surface hydrophobicity were increased after the sonication treatment indicating protein unfolding and exposure of the embedded SH and/or hydrophobic groups. An opposite trend was observed in HHP treated samples, implying aggregation and reassociation of structures under HHP. HHP and sonication treatments induced a decrease in ordered secondary structures (random coil and β-turn) accompanied with an increase in disordered secondary structures (α-helix and β-sheet) as probed by FTIR. Finally, the sonication treatment induced a significant improvement in the solubility (up to ∼3 folds at pH 7 and ∼2.6 folds at pH 9) and a reduction in particle sizes (up to ∼3 folds at pH 7 and ∼4.4 folds at pH 9). However, HHP treatment (600 MPa) only slightly increased the solubility (∼1.6 folds at pH 7 and ∼1.2 folds at pH 9) and decreased the particle size (∼1.3 folds at pH 7 and ∼1.2 folds at pH 9). This study provides a direct comparison of the impacts of sonication and HHP treatment on QPI, which will enable to choose the appropriate processing methods to achieve tailored properties of QPI.     

Evaluation of high hydrostatic pressure effects on bovine red blood cells and platelets

The objective of this study was to investigate the effects of high hydrostatic pressure (HHP) on the stability of red blood cells (RBCs) and platelets. Bovine blood cells (n=5) were treated with the pressure of 55, 110, 154 and 220 MPa at 25 °C for 5 min. Light microscopy, atomic force microscopy and flow cytometry studies revealed that RBCs were morphologically stable up until the 220 MPa pressure treatments, at which surface modifications were observed. The platelets were found to be less stable than RBCs. HHP application did not cause any significant change in the signal intensity, band area and frequency values of the infrared bands with the exception that a significant variation was observed in the area of the cholesterol band. No statistically significant variations were observed in the secondary structure elements due to HHP treatment according to the artificial neural network study based on the FTIR data.     

The effect of high hydrostatic pressure on the survival of Saccharomyces cerevisiae in model suspensions and beetroot juice

The inactivation of Saccharomyces cerevisiae NCFB 3191 using high hydrostatic pressure of 300 MPa at 20°C with a holding time of 0, 1, 5 and 10 min was investigated with model suspensions in phosphate-buffered saline and in beetroot juice. The reduction in S. cerevisiae NCFB 3191 in model suspensions was about 5 log after 10 min of pressurization, irrespective of the initial level of cell concentration in the samples (5.4–8.7 log cfu/mL). The baroprotective effect of beetroot juice on yeast cells during pressurization was observed; the reduction was lower and was only 3.5 log (the inoculum was 5.4 log cfu/mL). No sublethal injury among the surviving cells of the studied yeast strain was found.     

Quantitative evaluation of the microjet velocity and cavitation erosion on a copper plate produced by a spherical cavity focused transducer at the high hydrostatic pressure

Cavitation erosion at the high hydrostatic pressure causes the equipment to operate abnormally for the huge economic losses. Few methods can quantitatively evaluate the cavitation erosion intensity. In order to solve this problem, the cavitation erosion on a copper plate was carried out in a spherical cavity focused transducer system at the hydrostatic pressure of 3, 6, and 10 MPa. Meanwhile, the corresponding cavitation threshold, the initial bubble radius, and the microjet velocity in the ultrasonic field are theoretically analyzed to determine the dimension and velocity of microjet based on the following hypotheses: (1) the influence of the coalescence on the bubble collapse is ignored; (2) the dimension of the microjet is equal to the largest bubble size without the influence of gravity and buoyancy. Using the Westervelt equation for the nonlinear wave propagation and the Johnson-Cook material constitutive model for the high strain rate, a microjet impact model of the multi-bubble cavitation was constructed. In addition, through the analogy with the indentation test, an inversion model was proposed to calculate the microjet velocity and the cavitation erosion intensity. The microjet geometric model was constructed from the dimension and velocity of the microjet. The continuous microjet impact was proposed according to the equivalent impact momentum and solved by the finite element method. The relative errors of the pit depth are 4.02%, 3.34%, and 1.84% at the hydrostatic pressure of 3, 6, and 10 MPa, respectively, and the relative error in the evolution of pit morphology is 7.33% at 10 MPa, which verified the reliability of the proposed models. Experimental and simulation results show that the higher the hydrostatic pressure, the greater the pit depth, pit diameter, the pit-to-microjet diameter ratio, and the cavitation erosion intensity, but the smaller the pit diameter-to-depth ratio. The cavitation erosion intensity becomes significant with the ongoing ultrasonic exposure. In addition, a comparison of the cavitation pit morphology in the microjet pulsed and continuous impact modes shows that the continuous impact mode is effective without the elastic deformation caused by the residual stress. Using the cavitation pit morphology at the different hydrostatic pressures, the microjet velocity can be estimated successfully and accurately in a certain range, whose corresponding errors at the lower and upper limit are 5.98% and 0.11% at 3 MPa, 6.62% and 9.14% at 6 MPa, 6.54% and 5.42% at 10 MPa, respectively. Our proposed models are valid only when the cavitation pit diameter-to-depth ratio is close to 1. Altogether, the cavitation erosion induced by multi-bubble collapses in the focal region of a focused transducer could be evaluated both experimentally and numerically. Using the cavitation pit morphology and the inversion model, the microjet velocity in a certain range could be estimated successfully with satisfactory accuracy.     

High hydrostatic pressure inactivation of Lactobacillus plantarum cells in (O/W)-emulsions is independent from cell surface hydrophobicity and lipid phase parameters

Inactivation efficiency of high hydrostatic pressure (HHP) processing of food is strongly affected by food matrix composition. We investigated effects of fat on HHP inactivation of spoilage-associated Lactobacillus (L.) plantarum strains using defined oil-in-water (O/W)-emulsion model systems. Since fat-mediated effects on HHP inactivation could be dependent on interactions between lipid phase and microbial cells, three major factors possibly influencing such interactions were considered, that is, cell surface hydrophobicity, presence and type of surfactants, and oil droplet size. Pressure tolerance varied noticeably among L. plantarum strains and was independent of cell surface hydrophobicity. We showed that HHP inactivation of all strains tended to be more effective in presence of fat. The observation in both, surfactant-stabilized and surfactant-free (O/W)-emulsion, indicates that cell surface hydrophobicity is no intrinsic pressure resistance factor. In contrast to the presence of fat per se, surfactant type and oil droplet size did not affect inactivation efficiency.     

Overexpression of EAR1 and SSH4 that encode PPxY proteins in the multivesicular body provides stability to tryptophan permease Tat2, allowing yeast cells to grow under high hydrostatic pressure

Tryptophan uptake in yeast Saccharomyces cerevisiae is susceptible to high hydrostatic pressure and it limits the growth of tryptophan auxotrophic (Trp^?) strains under pressures of 15–25 MPa. The susceptibility of tryptophan uptake is accounted for by the pressure-induced degradation of tryptophan permease Tat2 occurring in a Rsp5 ubiquitin ligase-dependent manner. Ear1 and Ssh4 are multivesicular body proteins that physically interact with Rsp5. We found that overexpression of either of the EAR1 or SSH4 genes enabled the Trp^? cells to grow at 15–25 MPa. EAR1 and SSH4 appeared to provide stability to the Tat2 protein when overexpressed. The result suggests that Ear1 and Ssh4 negatively regulate Rsp5 on ubiquitination of Tat2. Currently, high hydrostatic pressure is widely used in bioscience and biotechnology for structurally perturbing macromolecules such as proteins and lipids or in food processing and sterilizing microbes. We suggest that hydrostatic pressure is an operative experimental parameter to screen yeast genes specifically for regulation of Tat2 through the function of Rsp5 ubiquitin ligase.     

On the nature and the practical relevance of the molecular modifications induced in wheat proteins by high pressure treatment

Abstract

Modifications induced by the combined application of pressure (up to 800 MPa) and temperature (up to 80° C) were studied on wet and dry semolina, and on fresh pasta as such and after cooking. Rheological and immunochemical properties of the treated products were investigated, along with parameters indicating the extent of protein denaturation. No modification occurs in semolina treated at less than 45% water, below 60°C/800MPa for 10min. Egg noodles prepared from pressure/heat treated semolina had increased protein digestibility and low recognition by anti-gliadin antibodies either before or after cooking. The same effects were observed in heat/pressure treated pasta, but were lost upon subsequent cooking, although we found decreased immunochemical reactivity after heat/pressure treatment of cooked pasta. No sensory difference was found between cooked and pressure-treated semolina products.     

Tetrahedral versus octahedral Mn site coordination in wurtzite and rocksalt Zn1−xMnxO investigated by means of XAS experiments under high pressure

We present the results of x-ray absorption measurements carried out in Zn_1−xMn_xO thin films under high pressure. The Mn environment remains essentially the same for nominal Mn concentrations given by x=0.05, 0.1, 0.15 and 0.25. Both the XANES (X-ray Absorption Near Edge Structure) and EXAFS (Extended X-ray Absorption Fine Structure) indicate that Mn occupies the Zn site, being surrounded by four oxygen atoms at 2.02±0.01 Å. The substitutional hypothesis is reinforced by comparing the differences between the ambient (wurtzite) and high pressure (rocksalt) spectra, which correspond to tetrahedral and octahedral Mn environments.     

Study of the bandgap renormalization in Ga-doped ZnO films by means of optical absorption under high pressure and photoelectron spectroscopy

In this paper we investigate the band gap renormalization in heavily Ga-doped ZnO thin films deposited by pulsed laser deposition on C$C$-plane sapphire and mica substrates. Thin films were studied by ultraviolet photoelectron spectroscopy and also by optical measurements under high pressure. The Fermi-level shift, as obtained from ultraviolet photoelectron experiments, exhibits a relatively small and positive shift (about 0.3 eV) with respect to the valence band for increasing electron concentrations up to 10²¹ cm⁻³. The optical gap exhibits a much larger increase (0.6 eV) for the same concentration range. Absorption measurements under pressure show that the pressure coefficient of the optical gap is correlated to the electron concentration in films, decreasing with increasing electron concentration. As a consequence, the contributions of band filling and band renormalization to the optical gap shift can be separated on the basis of the different pressure behavior of the physical parameters involved in each effect.