Moderate hydrostatic pressure–temperature combinations for inactivation of Bacillus subtilis spores

We report the effect of using moderate hydrostatic pressure, 40–140?MPa, at moderate temperature (38–58°C) to inactivate Bacillus subtilis spores in McIlvaine's citric phosphate buffer at pH 6. We have investigated several parameters: pressure applied, holding time, pressure cycling, and temperature. The kinetics of spore inactivation is reported. The results show that spore inactivation is exponentially proportional to the time the sample is exposed to pressure. Spore germination and inactivation occur at the hydrostatic pressures/temperature combinations we explored. Cycling the pressure while keeping the total time at high pressure constant does not significantly increase spore inactivation. We show that temperature increases spore inactivation at two different rates; a slow rate below 33°C, and at a more rapid rate at higher temperatures. Increasing pressure leads to an increase in spore inactivation below 95?MPa; however, further increases in pressure give a similar rate kill. The time dependence of the effect of pressure is consistent with the first-order model (R²?>?0.9). The thermal resistance values (Z_T) of B. subtilis spores are 30°C, 37°C, and 40°C at 60, 80, 100?MPa. The increase in Z_T value at higher pressures indicates lower temperature sensitivity. The pressure resistance values (Z_P) are 125, 125 and 143?MPa at 38°C, 48°C, and 58°C. These Z_P values are lower than those reported for B. subtilis spores in the literature, which indicates higher sensitivity at pressures less than about 140?MPa. We show that at temperatures <60°C, B. subtilis spores are inactivated at pressures below 100?MPa. This finding could have implications for the design of the sterilization equipment.     

Listeria monocytogenes cells injured by high hydrostatic pressure and their recovery in nutrient-rich or -free medium during cold storage

ABSTRACT

Cells of Listeria monocytogenes suspended in phosphate-buffered saline (PBS) were treated by high hydrostatic pressure (HHP; 500?MPa, 25°C, 10?min), diluted by ten folds using trypticase soy broth (TSB) or PBS, and stored at cold temperatures of 0–15°C. Viable cell count in TSB increased logarithmically close to the initial count at each storage temperature, while that in PBS increased temporarily and subsequently decreased to almost nondetectable level except the case at 15°C, where it showed logarithmic increase thereafter. Based on proliferation experiments where their healthy cells were inoculated to TSB or to PBS containing their heat-killed dead cells, it was suggested that increase in the viable count of HHP-treated cells in TSB and PBS could be ascribed to the recovery of colony forming ability and/or proliferation depending on the cold storage temperature.     

Modelling of inactivation kinetics of Escherichia coli and Listeria monocytogenes on grass carp treated by combining ultrasound with plasma functionalized buffer

Linear (first-order) and non-linear (Weibull, biphasic, and log-logistic) models were evaluated for predicting the inactivation kinetics of Escherichia coli and Listeria monocytogenes on grass carp treated by a novel technique (UPFB) combining ultrasound (US) with plasma functionalized buffer (PFB). Results showed that UPFB was more effective for inactivating bacteria when compared with individual applications of US or PFB with reductions of 3.92 and 3.70 log CFU/g for Escherichia coli and Listeria monocytogenes, respectively. Compared with the linear model, the three non-linear models presented comparable performances and were more suitable for describing the inactivation kinetics with superior adj-R² (0.962–0.999), accuracies (0.970–1.006) and bias factors (0.995–1.031), and by assessing the strengths of evidence, weights of evidence and evidence ratios for the models, the biphasic model was identified as the best fit model. The current study provided new insights into the effective evaluation of decontamination methods.     

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.     

Nanovesicle encapsulation of antimicrobial peptide P34: physicochemical characterization and mode of action on <Emphasis Type="Italic">Listeria monocytogenes</Emphasis>

Antimicrobial peptide P34, a substance showing antibacterial activity against pathogenic and food spoilage bacteria, was encapsulated in liposomes prepared from partially purified soybean phosphatidylcholine, and their physicochemical characteristics were evaluated. The antimicrobial activity was estimated by agar diffusion assay using Listeria monocytogenes ATCC 7644 as indicator strain. A concentration of 3,200 AU/mL of P34 was encapsulated in nanovesicles and stocked at 4 °C. No significant difference (p > 0.05) in the biological activity of free and encapsulated P34 was observed through 24 days. Size and PDI of liposomes, investigated by light scattering analysis, were on average 150 nm and 0.22 respectively. Zeta potential was −27.42 mV. There was no significant change (p > 0.05) in the physicochemical properties of liposomes during the time of evaluation. The liposomes presented closed spherical morphology as visualized by transmission electron microscopy (TEM). The mode of action of liposome-encapsulated P34 under L. monocytogenes cells was investigated by TEM. Liposomes appeared to adhere but not fuse with the bacterial cell wall, suggesting that the antimicrobial is released from nanovesicles to act against the microorganism. The effect of free and encapsulated P34 was tested against L. monocytogenes, showing that free bacteriocin inhibited the pathogen more quickly than the encapsulated P34. Liposomes prepared with low-cost lipid showed high encapsulation efficiency for a new antimicrobial peptide and were stable during storage. The mode of action against the pathogen L. monocytogenes was characterized.     

High pressure inactivation of relevant target microorganisms in poultry meat products and the evaluation of pressure-induced protein denaturation of marinated poultry under different high pressure treatments

In this study, the possibility of extending shelf life of marinated poultry meat products by high pressure processing was evaluated. Relevant spoilage and pathogenic strains were selected and used as target microorganisms (MOs) for challenge experiments. Meat and brine were inoculated with MOs and treated at 450 MPa, 4 °C for 3 min. The results of inactivation show a decreasing pressure tolerance in the series Lactobacillus > Arcobacter > Carnobacterium > Bacillus cereus > Brochothrix thermosphacta > Listeria monocytogenes. Leuconostoc gelidum exhibited the highest pressure tolerance in meat. A protective effect of poultry meat was found for L. sakei and L. gelidum. In parallel, the influence of different marinade formulations (pH, carbonates, citrates) on protein structure changes during a pressure treatment was investigated. Addition of sodium carbonate shows a protection against denaturation of myofibrillar proteins and provides a maximum water-holding capacity. Caustic marinades allowed a higher retention of product characteristics than low-pH marinades.     

Effect of high pressure treatment on liquid whole egg

In our tests, we artificially infected liquid whole egg samples with Salmonella enteritidis, Listeria monocytogenes, and Staphylococcus aureus bacteria, and then treated the samples in “Food Lab900” high hydrostatic pressure (HHP) instrument for 3–17 min at 200–400 MPa. Subsequently, the change of the viable cell count of the specific bacteria has been tested. In addition to the samples infected with various bacteria, non-infected samples were also treated in each test and the change in viable cell count, colour and viscosity of the samples upon the effect of the treatment. In summary, it can be concluded that in each test of our investigations, the viable cell count of S. enteritidis critical for egg products is reduced significantly, while the reduction of the total viable cell count was around two magnitudes. Additionally, based on our results, microbial destruction, reduction of enthalpy (denaturation of egg white) caused by the treatment at HPP, and colour change are primarily affected by the pressure level, while the changes in rheological properties are also significantly affected by the duration of high pressure treatment (p<0.05).     

Microorganisms baroinactivation of germinated mung bean (green gram) seeds

It is possible to describe the pressure degradation of microorganisms as being analogous to thermal inactivation. Equation for baroinactivation is derived from thermal death time (TDT) model D _p = D _pref 10^{(pref-p)/z _p} , where D _pref is decimal reduction time at the reference pressure (min), p _ref is the reference pressure (MPa), z _p is the pressure increase (MPa) required to reduce the D _p value by a factor of 10. This method was used for the calculation of baroinactivation model parameters of the total number of microorganisms of pressurized germinated mung bean (green gram) seeds (Vigna radiata (L.) Wilczek). Microbial contamination of germinated mung bean (green gram) seeds can be effectively decreased by treatment at a high pressure of 400 MPa, time of pressurization 5 min.     

High pressure inactivation of Clostridium botulinum type E endospores in model emulsion systems

Clostridium botulinum type E is a cold-tolerant, neurotoxigenic, endospore-forming organism, primarily associated with aquatic environments. High pressure thermal (HPT) processing presents a promising tool to enhance food safety and stability. The effect of fat on HPT inactivation of C. botulinum type E spores was investigated using an emulsion model system. The distribution of spores in oil-in-water (O/W) emulsions and their HPT (300–750?MPa, 45–75?°C, 10?min) inactivation was determined as a function of emulsion fat content (30–70% (v/v) soybean oil in buffer). Approximately 26% and 74% of the spores were located at the oil–buffer interface and the continuous phase, respectively. Spore inactivation in emulsion systems decreased with increasing oil contents, which suggests that the fat content of food plays an important role in the protection of C. botulinum type E endospores against HPT treatments. These results can be helpful for future safety considerations.     

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 high pressure on growth and bacteriocin production of Pediococcus acidilactici HA-6111-2

This study was aimed to investigate the effect of high pressure processing (HPP, 200–600?MPa) on the (i) survival of Listeria innocua and Pediococcus acidilactici HA-6111-2; (ii) production of bacteriocin bacHA-6111-2 and (iii) activity of bacteriocin against untreated and pressure-treated L. innocua cells. Inactivation of P. acidilactici was observed for pressures of >300?MPa. However, at this pressure level, L. innocua was more sensitive. Bacteriocin crude extract was pressure stable, with a decrease for pressures of ≥400 MPa. Pressures of ≤200?MPa did not affect bacteriocin production when compared with non-pressure-treated cells, whereas higher pressures caused a 2- to 4-fold decrease on the maximum level of bacteriocin production. Growth curves of P. acidilactici were fitted with the modified Gompertz model. The lag phase period depended on the magnitude of the pressure applied: there was a delay in the exponential phase as pressure increased and, as a consequence, in the beginning of bacteriocin production. Since P. acidilactici HA-6111-2 and its bacteriocin have shown resistance to pressures up to 300–400 MPa, they could be used in combination with HPP in order to improve food safety.     

Decrease in optical density as a results of germination of Alicyclobacillus acidoterrestris spores under high hydrostatic pressure

Alicyclobacillus acidoterrestris is a spore-forming bacterium, causing spoilage of juices. The spores of these bacteria have the ability to survive in the typical conditions used for thermal pasteurization. Therefore, the use of other techniques such as high hydrostatic pressure is considered for their inactivation. The effect of hydrostatic pressure of 200–500 MPa, at temperatures 4–50 °C for 15 min, on the dynamics of germination of A. acidoterrestris spores in apple juice and pH 4 buffer was studied. To estimate the share of germinated spores, the method of determining the optical density at a wavelength of 660 nm (OD₆₆₀) was used. Parameters of hydrostatic pressure treatment used in this work affected the dynamics of germination of A. acidoterrestris spores in apple juice, and the temperature had the greatest effect. The results indicate that nutrients present in apple juice can promote the germination of A. acidoterrestris spores.     

The effect of high hydrostatic pressure on the physiological and biochemical properties of pepper (Capsicum annuum L.) seedlings

High hydrostatic pressure is a non-thermal food processing technology, which also has several successful applications in different areas besides food processing. In this study, Capsicum annuum L. (pepper) seeds are subjected to 50, 100, 200 and 300?MPa pressure for 5?min at 25°C and the seedlings of HHP processed seeds are used to compare percentage of seed germination and biochemical properties such as chlorophyll a, b and a/b, proline content, total protein, carotenoid, malondialdehyde, glucose, fructose and phenolic compounds concentrations. As a result of the study, it was observed that there are remarkable changes in terms of biochemical properties especially for seedlings, whose seeds were pressurized at 200 and 300?MPa. More detailed studies are needed to put forward the mechanism behind the changes in biochemical properties.     

大气压冷等离子体射流灭活子宫颈癌Hela细胞

研究了大气压冷等离子体射流对子宫颈癌Hela细胞的灭活机制. 在倒置显微镜下观察不同等离子体处理条件下的细胞形态, 并通过中性红吸收测试定量测定各个条件下的细胞存活率. 将功率维持在18 W, 在900 mL/min 氩等离子体中添入氧气的百分含量分别为1%, 2%, 4% 和8%的条件下处理Hela细胞, 探讨活性气体氧气在惰性气体氩气中的百分含量对Hela癌细胞灭活效率的影响, 发现添加2%氧气时, 氩/氧等离子体灭活效果最佳, 处理180 s后细胞存活率可降至7%. 当继续添加氧超过2%时, 灭活效果逐渐减弱, 直至8%时, 其效果反而不如单纯氩等离子体. 通过测量等离子体发射光谱, 结果表明活性氧自由基在癌细胞灭活过程中可能起关键作用. 关键词： 大气压冷等离子体射流 Hela癌细胞 存活率 发射光谱     

Structural modification of swai-fish (Pangasius hypophthalmus)-based emulsions containing non-meat protein additives by ultra-high pressure and thermal treatments

Swai-fish emulsions containing fermented soybeans (thua nao and rice-koji miso) were pressurized at 600?MPa for 20 min or heated at 72°C for 30 min. The fish batters were blended with soy protein isolate (SPI) or whey protein concentrate (WPC) to stabilize the emulsions. The processed fish emulsions were then subjected to physical, chemical and microbiological examinations. The results of gel strength and water-holding potential showed that SPI addition yielded higher impact on these properties than WPC addition, which was also confirmed by the interactions between SPI and native fish proteins depicted by electrophoregrams. The frequency profiles suggested that the heated gels had a greater storage and loss moduli than pressurized gels, while pressurized WPC set-gel displayed larger loss tangent (the predominance of viscous moiety) than those pressurized SPI set-gel. High bacteria and spore counts of B. subtilis (residual of the thua nao) were observed in both pressurized and heated fish-based emulsions.     

Effect of pH and high pressure at sub-zero temperature on the viability of some bacteria

The objective of this study was to investigate the viability of Escherichia coli and Staphylococcus aureus in apple juice after treatment with high pressure at sub-zero temperature and during subsequent storage at 5 and 20 °C. The viability of E. coli and S. aureus cells suspended in the apple juice with a pH of 3.8 did not decrease considerably after pressure treatment at 193 MPa and?20 °C. However, viability losses occurred during storage of samples after pressure treatment. Living cells of both strains were not detected in pressurized samples of apple juice stored for 10 days at 20°C. The lethal effect was lower when the samples after pressure treatment were incubated at refrigerated temperature; the number of E. coli and S. aureus decreased by 6 log cycles when the juice was stored for 10 days at 5 °C.     

Combined effects of high pressure and sodium hydrogen carbonate treatment on beef: improvement of texture and color

We investigated the effect of combined high pressure and sodium hydrogen carbonate (NaHCO₃) treatment on the physical properties and color of silverside Australian beef. Meat samples were pressurized at 100–500 MPa and the water content, weight reduction, rupture stress, and meat color were determined. The water content of meat treated with NaHCO₃ and high pressure (300 MPa) reached a maximum of 70.1%. Weight reduction tended to decrease with high pressure treatment at 300 MPa. Meats treated with NaHCO₃ and high pressure at 400 MPa showed a>50% decrease in hardness. Whitening of the meat was reduced by the combined high pressure and NaHCO₃ treatment. Therefore, the combined high pressure and NaHCO₃ treatment is effective for improvement of beef quality.     

Effect of high hydrostatic pressure on the profile of proteins extracted from Betula pendula pollens

High hydrostatic pressure (HHP) has high success potential in pollen protein extraction, but its effect on pollen protein profiles has not been studied yet. The aim of this study is to put forward whether HHP processing causes a change in the protein profiles extracted from pollens or not. In this study, proteins extracted from Betula pendula pollens were studied at 100, 200 and 300?MPa at room temperature for 5?min. In addition, the efficiency of three different extraction solvents, namely phosphate buffer saline (PBS) buffer pH 7.5, trichloroacetic acid–acetone and Tris–HCl buffer pH 8.8, was also observed, and the results were compared with the conventional pollen protein extraction procedure. As a result, it is concluded that 200?MPa for 5?min has extracted similar amounts of protein compared with the conventional extraction method which lasted for 24?h, which lasted for 24 h. On the other hand, the application time for 200 MPa for 5 min is extremely shorter when it is compared to the conventional extraction method.     

Prior exposure to an attenuated <Emphasis Type="Italic">Listeria</Emphasis> vaccine does not reduce immunogenicity: pre-clinical assessment of the efficacy of a <Emphasis Type="Italic">Listeria</Emphasis>vaccine in the induction of immune responses against HIV

Background  

We have evaluated an attenuated Listeria monocytogenes (Lm) candidate vaccine vector in nonhuman primates using a delivery regimen relying solely on oral vaccination. We sought to determine the impact of prior Lm vector exposure on the development of new immune responses against HIV antigens.     

Influence of high power ultrasound on natural microflora,pathogen and lactic acid bacteria in a raw meat emulsion

Raw meat emulsions may have natural, spoilage and pathogenic microorganisms due to the origin and characteristics of this food matrix. All of these microorganisms must be minimized during industrial processing to make food consumption safe and meet quality regulations. Therefore, in this research, the effect of probe ultrasound on the inactivation of three kinds of microorganisms in a raw meat emulsion is evaluated. The microorganisms are: natural microflora NAM, Listeria monocytogenes LIS, and Lactobacillus delbrueckii LAC. A high-intensity probe ultrasound system was used, during 1.0, 2.5, 5.0, 7.5 and 10 min, with pulsed waves of 0.0, 10, 20 and 30 seg, and 200, 250, 300, 350 and 400 W of power. The interrelation between time, wave pulse cycle, and power factors was assessed. The results showed a positive linear independence effect in the treatments without wave pulse for each microorganism, and a quadratic interaction with the time and the ultrasound power for the inactivation of the three kinds of microorganisms. Besides, the desirability function for the inactivation reached up to 60% of the microbial population with the probe ultrasound treatment, with 10 min, a 7.56 s wave pulse and 400 W of power. Thus, these results could be useful to decide the incorporation of mild and emerging technologies in a meat industry line process.