A Gompertz Model Approach to Microbial Inactivation Kinetics by High-Pressure Processing Incorporating the Initial Counts,Microbial Quantification Limit,and Come-Up Time Effects

During come-up time (CUT), the time to reach a desired processing pressure, isobaric-isothermal conditions cannot be assumed in the estimation of kinetic parameters for the design of commercial high-pressure processing (HPP) treatments. Since CUT effects on microbial population, enzyme activity, and chemical concentration are often ignored, kinetic models incorporating the non-isobaric and non-isothermal conditions prevailing during CUT were the objective of this work. The analysis of peer-reviewed data on the HPP inactivation of bacteria (counts observations n = 919, 60 survival curves) and bacterial spores (n = 273, 12 curves) showed that a Gompertz model (GMPZ) approach is an effective alternative. The GMPZ parameter A was fixed as the difference between the initial population (log₁₀ N _o ) and the lower quantification limit of microbial counts (log₁₀ N _lim), while exponential equations were used to describe pressure effects on the lag time (λ) and the maximum inactivation rate (μ_max). In low-acid media (pH > 4.5), λ decreased exponentially with pressure, allowing the identification of a theoretical pressure level (P _λ) sufficient to initiate microbial inactivation during CUT. The parameter μ_max exponentially increased with pressure for all evaluated datasets. Dynamic pressure effects during CUT were simplified by assuming isobaric conditions during CUT (t _CUT), allowing to obtain GMPZ parameter estimates using only nonlinear regression (R ² ～ 0.938, σ ² = 0.030–0.604). The proposed approach is a simpler, promising tool for a more informative analysis of the kinetics of microbial inactivation by HPP and should be further validated with additional experimental data.     

Effect of high‐pressure treatments on microbial loads and physicochemical characteristics during refrigerated storage of raw milk Serra da Estrela cheese samples

This work studied the effect of high pressure processing (HPP) at 400, 500 and 600 MPa during 10, 5 and 3 min, respectively, on samples ewe cheese manufactured from raw milk, during storage (100 days) at 5 °C. Total aerobic mesophilic and lactic acid bacteria were slightly affected, decreasing by about 1.0 and 0.82 log CFU g^?1, respectively, immediately after HPP treatment at 600 MPa for 3 min, while Enterobacteriaceae, yeasts and moulds, and Listeria innocua were reduced to below the quantification limits. Lactic acid bacteria decreased further during storage, showing increasing inactivation as the pressure level increased. Physicochemical parameters (water activity, moisture content, pH and titratable acidity) were generally not affected by HPP, while lipid oxidation increased throughout storage, with HPP samples showing lower values (50–66%) at 100 days of storage. The results indicated that HPP has potential to improve cheese microbial safety and shelf‐life, with a lower lipid oxidation level than nonpressurised cheese.     

Synergistic effects of high pressure processing and slightly acidic electrolysed water on the inactivation of Bacillus cereus spores

Bacillus spores are concerns for their resistance to heat, high pressure processing (HPP), and disinfectants. We examined the effects of HPP and slightly acidic electrolysed water (SAEW) on inactivation of B. cereus spores. Spores' suspensions were prepared with 2‐(N‐morpholino) ethanesulfonic acid (MES) buffer or SAEW with available chlorine content (ACC) of 24, 35, 44 or 55 mg L^?1, and then subjected to HPP. The individual effects of HPP or SAEW on spores were negligible (<1.0 log CFU mL^?1). With factorial design and anova analysis, HPP + SAEW treatment was shown to have significantly positive effects on spores’ inactivation. The optimal conditions were 300 MPa HPP + SAEW with 44 mg L^?1 ACC or 200 MPa HPP + SAEW with 44 mg L^?1 ACC + 500 MPa HPP, producing reductions of 3.27 and 3.99 log CFU mL^?1, respectively. HPP + SAEW have potentials to serve as two effective hurdle techniques for inactivating B. cereus spores.     

Effect of high-pressure processing on Listeria spp. and on the textural and microstructural properties of cold smoked salmon

Investigation of the effect of high-pressure processing (HPP) at very short time on the inactivation of Listeria innocua was conducted as well as the effect on texture and microstructure. Lipid oxidation, colour and background bacterial flora were studied as well. HPP at 700-900 MPa for 10 s increased the inactivation of L. innocua in cold smoked salmon from 4500 cfu/g to nondetectable level (<0.3 cfu/g). L. innocua was more sensitive to HPP than the background flora tested. The product presented good microbiological quality and there was no indication of lipid oxidation. The effect of HPP on the redness of the product was not observed, however immediate effect on the lightness was noticed and the salmon becomes lighter in colour as a function of both time and pressure. The effects on the microstructure increased with both time and pressure and were most significant at 900 MPa and 60 s. The effect on microstructure coincides with the reduction of the bacteria. The knowledge from this study provides information for the industry on the development of HPP at 400-900 MPa with short pressure time of less than 60 s.     

Effects of Water Activity in Model Systems on High-Pressure Inactivation of Escherichia coli

The objectives of the study were to measure the effect of water activity (a _w) and to quantitatively evaluate the effect of the selected humectants under high-pressure processing (HPP) in combination with processing parameters such as treatment time, temperature, and pressure on the inactivation of Escherichia coli K12 in solid and liquid model systems. Glycerol was used in liquid and solid models to vary a _w at 0.90, 0.95, and 0.99 levels. The model systems samples and transmitting media were preconditioned to initial temperatures of 4 and 20 °C to compensate for adiabatic heating upon compression to ensure that HPP treatments at 400 and 600 MPa were performed at final temperatures not higher than 40 °C. Decrease of a _w from 0.99 to 0.90 in glycerol-based models caused considerably less inactivation of E. coli K12 at tested pressures and temperatures. Effect of different humectants at a _w 0.95 and 0.99 on the inactivation of E. coli K12 was studied comparing glycerol, fructose, sodium chloride, and sorbitol. Among four types of solutes tested in the study, sodium chloride appeared the least protective, with glycerol and fructose being approximately equal, and sorbitol showed the most protective effects on inactivation of E. coli K12. The obtained data of E. coli K12 inactivation by HPP at varied a _w levels in different solutes demonstrated similar effects of a _w on microbial inactivation by thermal treatments. The results must be taken into account when HP preservation process and foods are developed. Mention of trade names and commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the National Center for Food Safety and Technology.     

Equivalent processing for pasteurization of a pineapple juice–coconut milk blend by selected nonthermal technologies

Identifying equivalent processing conditions is critical for the relevant comparison of food quality attributes. This study investigates equivalent processes for at least 5-log reduction of Escherichia coli and Listeria innocua in pineapple juice–coconut milk (PC) blends by high-pressure processing (HPP), pulsed electric fields (PEF), and ultrasound (US) either alone or combined with other preservation factors (pH, nisin, and/or heat). The two blends (pH 4 and 5) and coconut milk (pH 7) as a reference were subjected to HPP at 300–600 MPa, 20°C for 0.5–30 min; PEF at an electric field strength of 10–21 kV/cm, 40°C for 24 µs; and US at 120 µm amplitude, 25 or 45°C for 6 or 10 min. At least a 5-log reduction of E. coli was achieved at pH 4 by HPP at 400 MPa, 20°C for 1 min; PEF at 21 kV/cm, 235 Hz, 40°C for 24 µs; and US at 120 µm, 45°C for 6 min. As L. innocua showed greater resistance, a synergistic lethal effect was provided at pH 4 by HPP with 75 ppm nisin at 600 MPa, 20°C for 5 min; PEF with 50 ppm nisin at 18 kV/cm, 588 Hz, 40°C for 24 µs; and US at 45°C, 120 µm for 10 min. The total soluble solids (11.2–12.4°Bx), acidity (0.47%–0.51% citric acid), pH (3.91–4.16), and viscosity (3.55 × 10⁻³–4.0 × 10⁻³ Pa s) were not significantly affected under the identified equivalent conditions. HPP was superior to PEF and US, achieving higher ascorbic acid retention and lower color difference in PC blend compared to the untreated sample.     

Original article: Modelling the effects of food ingredients and pH on high‐pressure processing inactivation of Bacillus cereus spores: a laboratorial study

The combination of high pressure and heat on Bacillus cereus spores in food matrix was investigated with the purpose of achieving a predictive model of microbial inactivation. The high‐pressure processing (HPP) conditions were fixed at 540 MPa and 71 °C for 16.8 min, which were determined as the optimum conditions considering six‐log‐cycle reductions of B. cereus spores. The effects of soybean protein, sucrose, soybean oil and pH on the inactivation of B. cereus spores by HPP were evaluated, and a quadratic predictive model for the effects of food ingredients and pH on the reductions of B. cereus spores by HPP was built using response surface methodology. The experimental results showed that soybean protein, sucrose and pH significantly affected the reductions of B. cereus spores. The predictive model is significant, because the level of significance was P < 0.0001 and the calculated F‐value is much greater than the tabulated F‐value. The adequacy of the predictive model equation was verified effectively using the experimental test data that were not used in the development of the model.     

Supercritical CO2 and N2O pasteurisation of peach and kiwi juice

The microbial inactivation and qualitative parameters (pH, sugar content, titratable acidity, absorbance at 420 nm and turbidity) of peach and kiwi juices treated at 35 °C with supercritical carbon dioxide (SC‐CO₂) and nitrous oxide (SC‐N₂O) were determined as a function of pressure and treatment time. Total inactivation of both naturally occurring microorganisms and Saccharomyces cerevisiae strain (10⁵ cfu mL^?1) was obtained after 15 min of SC‐CO₂/N₂O treatment, 10 MPa and 35 °C, for both juices. No significant changes in chemical‐physical or in sensorial characteristics between untreated and treated juice were detected. The results obtained demonstrate the feasibility and the potential of SC‐CO₂/N₂O treatment as an alternative low temperature pasteurisation process for peach and kiwi juices.     

Factors affecting the resistance of listeria monocytogenes to high hydrostatic pressure

Several factors relevant to food preservation had large effects on the resistance of L. monocytogenes to high hydrostatic pressure. Cells in the stationary phase of growth were much more resistant to pressures above 200MPa than those in the exponential phase. Following treatment for 10 min at 400 MPa viable numbers of stationary phase cells were reduced by only 1.3 log₁₀ units compared with more than 7 log₁₀ units for exponentially growing cells. Stationary phase cells were sensitised to pressure by butylated hydroxyanisole, potassium sorbate and by acid conditions. The most effective sensitiser was BHA which, at 1.55 mM, caused a 10⁴‐fold enhancement of killing by treatment at 300 MPa for 10 min. Propyl hydroxybenzoate, butylated hydroxytoluene and sodium ascorbate were without effect. Xylitol (15%) protected log phase cells against inactivation by pressure: after treatment at 350 MPa for 10 min viable numbers were reduced by only 0.24 log₁₀ units compared with 2.4 log₁₀ units in the absence of xylitol. The physicochemical environment can substantially increase or decrease resistance to pressure and these effects will require quantification if the microbiological effects of pressure are to be predicted.     

Effect of High-Pressure Processing on Vibrio parahaemolyticus Strains in Pure Culture and Pacific Oysters

Different strains of Vibrio parahaemolyticus (Vp) in broth cultures and Vp‐inoculated live Pacific oysters (Crassostrea gigas) were subjected to high‐pressure processing (HPP) at 241, 276, 310, and 345 MPa. Results showed Vp numbers were reduced by HPP in both pure culture and whole oysters. Vp inactivation was dependent on time and pressure. Optimum conditions for reducing Vp in pure culture and oysters to nondetectable levels were achieved at 345 MPa for 30 and 90 s, respectively. Resistance variations were detected between Vp in pure culture and in oysters. HPP proved to be an efficient means of reducing Vp in oysters.     

High‐pressure processing‐induced conformational changes during heating affect water holding capacity of myosin gel

This study aimed to investigate the effects of high‐pressure processing (HPP) (0.1‐400 MPa for 9 min) on the water holding capacity (WHC) of heat‐induced rabbit myosin gel and structural changes during thermal treatment (25–75 °C). HPP at 100 MPa significantly increased the WHC (P < 0.05) and formed more regular and homogeneous three‐dimensional network. Myosin tails at 100 MPa unfolded completely during the thermal treatment, which was beneficial to form a high WHC gel network. However, myosin pressurised at 200 MPa and above formed a weak gel. Their heads were already aggregated before heating, preventing from subsequent thermal denaturation and aggregation. With the temperature increasing, unfolding of myosin tails was not sufficient for a filamentous network formation. These results suggested that HPP could modify the myosin structure and affect the gel formation during heating. The 100 MPa was the optimum pressure level for the WHC of rabbit myosin gel.     

Effect of high hydrostatic pressure and high-pressure homogenisation on <Emphasis Type="Italic">Lactobacillus plantarum</Emphasis> inactivation kinetics and quality parameters of mandarin juice

The inactivation kinetics of Lactobacillus plantarum in a mandarin juice treated by thermal treatment (45–90 °C), high-pressure homogenisation (HPH) (30–120 MPa at 15 and 30 °C) and high-pressure processing (HPP) (150–450 MPa at 15, 30 and 45 °C) were fitted to different Weibullian equations. A synergic effect between pressure and temperature was observed in HPH and HPP treatments achieving 2.38 log cycles after 120 MPa at 30 °C for 10 s (final T of 45 °C) and 6.12 log cycles after 400 MPa at 45 °C for 1 min (final T of 60 °C), respectively. A combined treatment of 100 MPa at 15 °C for 10 s and 300 MPa at 15–30 °C for 1 min in HPH and HPP, respectively, was needed to the first logarithm microbial population decline. Weibull model accurately predicted microorganism inactivation kinetics after HPH and HPP processing when displaying single shoulder or tail in the survivor curves, whereas when a more complex trend was observed after thermal treatment, the double-Weibull equation was found more appropriate to explain such behaviour. Equivalent treatments that achieved the same degree of microbial inactivation (77 °C–10 s in thermal processing, 120 MPa–10 s at 30 °C in HPH processing and 375 MPa–1 min at 30 °C in HPP) were selected to study the effects on quality parameters. The application of dynamic pressure led to a decrease in sedimentable pulp, transmittance and juice redness, thus stabilising the opaqueness and cloudiness of mandarin juice. Pectin methyl esterase (PME) was found to be highly baroresistant to static and dynamic pressure. Carotenoid content remained unaffected by any treatment. This study shows the potential of high-pressure homogenisation as an alternative for fruit-juice pasteurisation.     

Evaluation of plasma,high‐pressure and ultrasound processing on the stability of fructooligosaccharides

Fructooligosaccharides (FOS) are among the main carbohydrates with prebiotic activity, and they are the most applied functional carbohydrate ingredient in the food industry. FOS are known to hydrolyse when subjected to thermal processing, thus partially losing its functional properties. In this study, we evaluate whether three nonthermal technologies are suitable for processing FOS regarding its stability after processing. FOS were subjected to ultrasound, high‐pressure processing (HPP) and atmospheric cold plasma (ACP). The FOS solution, 70 g L^?1, was set at a concentration recommended for human intake. The treatments were carried out at operating conditions usually used for microbial inactivation in foods (HPP at 450 MPa for 5 min; US at 600–1200 W L^?1 for 5 min; ACP at 70 kV for 15–60 s). NMR and HPLC analysis of the FOS components showed that ACP, ultrasound and HPP have not induced any significant change on FOS concentration (<2.0%) nor on the degree of polymerisation of the FOS (<3.3%). Contrarily to what is reported for thermal treatments, these nonthermal technologies were considered suitable for FOS processing.     

Modeling the inactivation of Salmonella typhimurium by dense phase carbon dioxide in carrot juice

The inactivation of Salmonella typhimurium inoculated into acidified carrot juice subjected to dense phase carbon dioxide (DPCD) was investigated. The pressures in the study were 10, 20 and 30 MPa, the temperatures were 32, 37 and 42 °C, and the treatment time was 5–90 min. The inactivation effect of DPCD was enhanced by increasing pressure and temperature. The sigmoid inactivation curves were characterized with the lag phase, exponential inactivation phase, and resistant phase. The inactivation curves were fitted to the modified Gompertz equation and the modified Logistic equation, the modified Gompertz equation was superior since its lowest residual sum of squares (RSS) was lower although there was no significant difference of goodness-of-fit between both models as indicated by F-test. The λ (the duration of the lag phase) and t_4-D (the time necessary to achieve 4-log cycles reduction) decreased with increasing pressure or temperature. The k_dm (the maximum specific value of the inactivation rate, min⁻¹) increased with increasing temperatures, and decreased with increasing pressures. The activation energy (Ea) and the activation volume (Va) necessary for inactivating S. typhimurium by DPCD were 19.06–29.39 kJ mol⁻¹ and 18.89–58.27 cm³ mol⁻¹.     

Residual polyphenol oxidase and peroxidase activities in high pressure processed bok choy (Brassica rapa subsp. chinensis) juice did not accelerate nutrient degradation during storage

Polyphenol oxidase (PPO) and peroxidase (POD) cannot be fully inactivated by commercial high pressure processing (HPP) operations, and their residual activities may accelerate nutrient degradation during storage. This study hence aimed to establish the effect of residual enzyme activity on nutrient preservation in bok choy (Brassica rapa subsp. chinensis) juice. Bok choy juice was treated at 600 MPa for up to 20 min and enzyme inactivation, nutrient retention immediately after treatment and nutrient preservation during storage were determined. High residual PPO (85.1 ± 2.6%) and POD (68.5 ± 1.0%) activities remained after 20 min of treatment. Increasing the pressure holding time to enhance enzyme inactivation did not compromise total antioxidant capacity, vitamin C, carotenoids, isothiocyanates and vitamin K levels. Neither did it significantly reduce the vitamin C degradation rate during refrigerated storage. Maximising enzyme inactivation may thus not be necessary for nutrient preservation during the storage of HPP-treated bok choy juice.Industrial relevance textWith HPP, an increase in pressure or holding time is required to achieve higher levels of enzyme inactivation. Without the need to maximize PPO and POD inactivation, juice processors may employ the minimum pressure and holding time required for microbial inactivation. As vegetative bacteria are typically less resistant to HPP inactivation than these enzymes, this could translate to reduced energy costs and increased throughput.     

Modeling the protective effect of aw and fat content on the high pressure resistance of Listeria monocytogenes in dry-cured ham

High pressure processing (HPP) is a promising food preservation technology as an alternative to thermal processing for microbial inactivation. The technological parameters, the type of microorganism, and the food composition can greatly affect the microbicidal potential of HPP against spoilage and pathogenic microorganisms. Presently, the number of available models quantifying the influence of food characteristics on the pathogen inactivation is scarce. The aim of this study was to model the inactivation of Listeria monocytogenes CTC1034 in dry-cured ham, as a function of pressure (347–852 MPa, 5 min/15 °C), water activity (a_w, 0.86–0.96) and fat content (10–50%) according to a Central Composite Design. The response surface methodology, based on the equation obtained with a stepwise multivariate linear regression, was used to describe the relationship between bacterial inactivation and the studied variables. According to the best fitting polynomial equation, besides pressure intensity, both a_w and fat content exerted a significant influence on HP-inactivation of L. monocytogenes. A clear linear piezoprotection trend was found lowering the a_w of the substrate within the whole range of tested pressure. Fat content was included in the model through the quadratic term and as interaction term with pressure, resulting in a particular behavior. A protective effect due to the presence of high fat content was identified for pressure treatments above ca. 700 MPa. At lower pressure, higher inactivation of L. monocytogenes occurred by increasing the fat content above 30%. The results emphasize the relevant influence of intrinsic factors on the L. monocytogenes inactivation by HPP, making necessary to assess and validate the effectiveness of HPP on specific food products and consequently set process criteria adjusted to each particular food product.     

Comparison of hydrostatic and hydrodynamic pressure to inactivate foodborne viruses

The effect of high hydrostatic pressure (HPP) and hydrodynamic pressure (HDP), in combination with chemical treatments, was evaluated for inactivation of foodborne viruses and non-pathogenic surrogates in a pork sausage product. Sausages were immersed in distilled water, 100-ppm EDTA, or 2% lactoferrin, and then inoculated with feline calicivirus (FCV), hepatitis A virus (HAV) or bacteriophage (MS2, phiX174, or T₄). Each piece was packaged individually and subjected to pressure by either HDP, HPP (500 MPa, 5 min, 4 °C), or control (no pressure). On sausages immersed in water, HPP and HDP significantly (P < 0.05) reduced titers of FCV by 2.89 and 2.70 log₁₀ TCID₅₀/ml, and HAV by log₁₀ 3.23 and 1.10, respectively, when compared to non-pressure-treated controls. Titers of T₄ (1.48 and 1.10 log₁₀ PFU/g) and MS2 (1.46 and 0.96 log₁₀ PFU/g) were also significantly reduced by HPP and HDP treatments, respectively, in combination with water. Inoculation of viruses and bacteriophage on a meat product may have protected viruses from complete inactivation by pressure treatments.

Industrial relevance

This is the first study to directly compare hydrostatic and hydrodynamic pressure technologies to inactivate microorganisms. This is also the first study to examine the inactivation of viruses and bacteriophages by pressure technology in a deli meat product. This study shows that viruses attached to meat surfaces may be protected from complete inactivation by hydrostatic and hydrodynamic pressure treatments, and these findings require more investigation into the survival of viruses in deli meat products.     

Interaction of β‐casein with (−)‐epigallocatechin‐3‐gallate assayed by fluorescence quenching: effect of thermal processing temperature

The interactions between the flavan‐3‐ol (?)‐epigallocatechin‐3‐gallate (EGCG) and bovine β‐casein in phosphate‐buffered saline (PBS) of pH 6.5 subjected to thermal processing at various temperatures (25–100 °C) were investigated using fluorescence quenching. The results indicated that different temperatures had different effects on the structural changes and EGCG‐binding ability of β‐casein. At temperatures below 60 °C, the β‐casein–EGCG interaction changed little (P > 0.05) with increasing temperature. At temperatures above 80 °C, native assemblies of β‐casein in solution dissociated into individual β‐casein molecules and unfolded, as demonstrated by a red shift of the maximum fluorescence emission wavelength (λ_max) of up to 8.8 nm. The highest quenching constant (K_q) and the number of binding sites (n) were 0.92 (±0.01) × 10¹³ m ^?1 s^?1 and 0.73 (±0.02) (100 °C), respectively. These results provide insight into the potential of interactions between β‐casein–EGCG that may modulate bioactivity or bioavailability to be altered during thermal process.     

Mathematical modeling of Saccharomyces cerevisiae inactivation under high‐pressure carbon dioxide

High‐pressure carbon dioxide inactivation curves of Saccharomyces cerevisiae at different temperatures were analysed using the modified Gompertz model. Comparable λ and μ values were obtained under pressure treatment as function of temperature. The phase of disappearance (λ) and the inactivation rate (μ) of S. cerevisiae were inversely related. Higher μ values were obtained at 50°C than at 40, 30, and 20°C under 10.0 MPa CO₂ pressure. Increased pressure and temperature had significant effects on the survival of S. cerevisiae. Arrhenius, linear and square‐root models were used to analyse the temperature dependence of the inactivation rate constant. For the Arrhenius model the activation energy (E_μ) was 56.49 kJ/mol at 10.0 MPa, and 55.70, 53.83, and 52.20 kJ/mol at 7.5, 5.0, and 2.5 MPa, respectively. Results of this study enable the prediction of yeast inactivation exposed to different CO₂ pressures and temperatures.     

HIGH PRESSURE DESTRUCTION KINETICS of INDIGENOUS MICROFLORA and ESCHERICHIA COLI IN RAW MILK AT TWO TEMPERATURES

Raw milk with a high count of indigenous microflora, with and without inoculated Escherichia coliK‐12 (ATCC‐29055), was subjected to high pressure treatment (250–400 MPa) for various holding times (0–80 min) at two temperatures (3 and 21C). From the microbial survivor data, an instantaneous pressure kill value (IPK) which represented the effect of a pressure pulse, and kinetic parameters (rate constant, D‐value, z‐value and activation volume) were evaluated based on pressure hold‐time first order kinetics. Both IPK and D values were dependent on pressure level and temperature. Higher pressures, longer holding times and lower temperatures resulted in larger destruction of microorganisms, and E. coli was more pressure sensitive than indigenous microflora. Pressure dependency of kinetic parameters was well described by both pressure death time and Eyring‐Magee models. At 3 and 21C, the z_p values were 227 and 240 MPa and λV# values were ?2.32 and ?2.34 (x 10^?5 m³ mole^?1), respectively, for indigenous microflora, and 179 and 205 MPa (z_p) and ?2.95 and ?2.75 (x 10^?5 m³ mole^?1) (λV#), respectively, for E. coli.