Clostridium sporogenes-ATCC 7955 Spore Destruction Kinetics in Milk Under High Pressure and Elevated Temperature Treatment Conditions

Clostridium sporogenes (ATCC 7955) spores inoculated in milk (2% fat) were subjected to high-pressure (HP) treatments (700–900 MPa) and at elevated temperatures (80–100 °C) for selected times up to 32 min. Samples were sealed in 1-mL plastic vials and placed in a specially constructed insulated chamber to prevent temperature drop during the treatment. Both pressure pulse (with no hold time) and pressure hold techniques were employed for treatment. Pressure pulse resulted in a small, but consistent, destruction (up to 0.5 log kill) of spores. During the pressure hold treatment, the destruction followed a first-order model (R ² > 0.90). The kinetic data were compensated for the small variations in temperature during the treatment. As expected, higher pressures and higher temperatures resulted in a faster rate of spore destruction. Temperature-corrected D values ranged from 13.6 to 2.4 min at 700 MPa and 7.0 to 1.3 min at 900 MPa, respectively, with process temperatures set at 90 and 100 °C. In comparison, thermal treatments gave D values ranging from 156 min at 90 °C to 12.1 min at 100 °C. The temperature sensitivity Z _P values (16.5 to 20.3 °C) under high pressure (700–900 MPa) were higher than under conventional thermal processing (9.0 °C), indicating the spore’s thermal resistance to increase at HP processing conditions. The pressure sensitivity Z _T values varied between 450 and 680 MPa under the elevated temperature (80–100 °C) processing conditions. Overall, C. sporogenes 7955 spores were relatively more sensitive to temperature than pressure.     

Characterization of metastable high pressure phase transition positions and its influence on the behavior of microbial destruction

Effect of perturbation factors on phase transition metastable positions of whole milk (4% fat content) and their influence on microbial destruction characteristics of non-pathogenic Escherichia coli inoculated in milk subjected to high pressure low temperature treatment were evaluated using a specially developed high pressure (HP) cooling system. Initially, the phase transition data of milk transitioning through the metastable phases were obtained and fitted successfully using Simon-like models as done in previous studies and polynomial formulas with R² of 0.997 & 0.996 for ice I, and 0.989 & 0.989 for ice III, respectively. The phase transition position of milk was explored with 5% and 10% sodium chloride solution as perturbation sources, respectively. Results showed that the 5% sodium chloride solution can reduce the transition pressure of milk by 43 MPa and increase the transition temperature by 4.1 °C, so that the milk can achieve phase transition at lower pressure and higher temperature. Phase transition microbial destruction was characterized by discontinuity, mutation and segmentation when the phase transition pressure interval 250– 300 MPa was carefully refined. The inactivation amount of E. coli before the phase transition (250 MPa) was 1.11 log and the phase transition process itself brought an additional 1.26 log destruction of E. coli population in milk.Industrial relevanceHigh pressure low temperature (HPLT) phase change kinetics were employed to enhance microbial destruction. HPLT was established based on a self-cooling unit positioned inside conventional HP chamber offering opportunities for scale up and commercialization. The effectiveness of HPLT phase transition for Escherichia coli destruction was demonstrated. The related research in metastable state provides a reference point for commercial application of high-pressure-low-temperature technology for microbial destruction and quality enhancement.     

High-pressure destruction kinetics of Clostridium sporogenes ATCC 11437 spores in milk at elevated quasi-isothermal conditions

The high-pressure sterilization establishment requires data on isobaric and isothermal destruction kinetics of baro-resistant pathogenic and spoilage bacterial spores. In this study, Clostridium sporogenes 11437 spores (10⁷ CFU/ml) inoculated in milk were subjected to different pressure, temperature and time (P, T, t) combination treatments (700–900 MPa; 80–100 °C; 0–32 min). An insulated chamber was used to enclose the test samples during the treatment for maintaining isobaric and quasi-isothermal processing conditions. Decimal reduction times (D values) and pressure and temperature sensitivity parameters, Z_T (pressure constant) and Z_P (temperature constant) were evaluated using a 3 × 3 full factorial experimental design. HP treatments generally demonstrated a minor pressure pulse effect (PE) (no holding time) and the pressure hold time effect was well described by the first order model (R² > 0.90). Higher pressures and higher temperatures resulted in a higher destruction rate and a higher microbial count reduction. At 900 MPa, the temperature corrected D values were 9.1, 3.8, 0.73 min at 80, 90, 100 °C, respectively. The thermal treatment at 0.1 MPa resulted in D values 833, 65.8, 26.3, 6.0 min at 80, 90, 95, 100 °C respectively. By comparison, HP processing resulted in a strong enhancement of spore destruction at all temperatures. Temperature corrected Z_T values were 16.5, 16.9, 18.2 °C at 700, 800, 900 MPa, respectively, which were higher than the thermal z value 9.6 °C. Hence, the spores had lower temperature sensitivity at elevated pressures. Similarly, corrected Z_P values were 714, 588, 1250 MPa at 80, 90, 100 °C, respectively, which illustrated lower pressure sensitivity at higher temperatures. By general comparison, it was concluded that within the range operating conditions employed, the spores were relatively more sensitive to temperature than to pressure.     

Melting endothermic technique for establishing different phase diagram pathways during high pressure treatment of liquid foods

The principle of endothermic melting was used to trace the path of phase diagram of foods covering Ice I and Ice III. Fitting the experimental data by parameters of the Simon-Like equations for pure water, R² values of 0.999 and 0.952 for Ice I and Ice III, respectively, were obtained with a mean error less than 5%. Using this method, phase diagrams of fruit juice and milk in Ice I and Ice III were then obtained, and R² values in Ice I and Ice III of 0.990 & 0.583 for fruit juice, and 0.991 & 0.886 for milk, respectively, were obtained. Similarly, based on the principle of the melting endothermic of glucose solution and sodium chloride solution under high pressure, pure water and milk could be cooled to pass through different phase transitions regimes. The developed concept and experimental set up are currently can be used for evaluating microbial destruction kinetics as influenced by phase transition HP treatment.Industrial relevanceThis paper uses the principle of melting endotherm to experimentally obtain the phase diagram of juice and milk, and realize the self-cooling of water and milk. The food (water and milk) can be cooled by the melting endotherm of cooling solution using a simple cooling set-up, and achieve different phase transition profiles with any ordinary high pressure treatment vessel that can operate at 400 MPa. This method overcomes the limitation of previous research studies which rely on more sophisticated and expensive high-pressure- equipment with jacketing and cooling of the pressure chamber with an add on refrigeration source. The developed technique can be used to evaluate microbial destruction as influenced by phase transition, greatly simplifying the experimental process and equipment requirement.     

HIGH PRESSURE–HIGH TEMPERATURE STERILIZATION: FROM KINETIC ANALYSIS TO PROCESS VERIFICATION†

The inactivation of Clostridium sporogenes PA 3679 spores by high pressure at high temperatures (HP–HT) in phosphate buffer was investigated in a lab‐scale temperature‐controlled HP system (QFP‐6) with an internal heater to maintain the sample temperature. Some inactivation of spores occurred during the pressurization come‐up time (CUT) and depressurization time. The inactivation of PA 3679 was found to be exponential during the adiabatic holding period of the HP cycle at constant pressures and temperatures. The inactivation rate increased with both pressure and temperature. The kinetic parameters – such as D‐values at tested temperatures and pressures that are necessary for the design of process parameters of HP sterilization process – were determined. Within the pressure range of 600–800 MPa, the calculated D‐values ranged from 270.3 to 357.4 and 49.0 to 67.6 s at 91 and 108C, respectively. These studies provided basic data on the effects of pressure and temperature on the inactivation of PA 3679 spores under conditions applicable to the development of preservation specifications for commercial HP–HT processing of low acid foods. The spore strips of C. sporogenes were used as indicators for microbiological verification of delivered lethality of HP–HT sterilization process at different processing conditions in a pilot scale HP vessel.     

Cycling versus Continuous High Pressure treatments at moderate temperatures: Effect on bacterial spores?

The advantage of using high pressure (HP) cycling treatment compared with continuous HP treatment was investigated for the inactivation of bacterial spores. The effects of parameters such as pulse number, pressure level, treatment temperature, compression and decompression rates, and time between pulses were evaluated. For this purpose, Bacillus subtilis and B. cereus spores (10⁸ and 10⁶ CFU/mL respectively) were suspended in 2-(N-morpholino) ethanesulfonic acid (MES) buffer solution, tryptone salt (TS) buffer solution, or infant milk and treated by HP cycling at 300–400 MPa, at 38–60 °C, for 1–5 pulses. Pressure cycling reduced the number of viable spores by 1.8 and 5.9 log respectively for B. subtilis and B. cereus species. Continuous HP treatments were performed at the same pressure and temperature for similar treatment durations. Our results showed that the spore inactivation ratio was correlated with the cumulative exposure time to pressure rather than to effects of the cycling process. Greater spore inactivation caused by HP cycling was observed only when faster compression and decompression rates were applied, probably due to adiabatic heating. A three-step kinetic model was developed, which seemed to support our hypothesis regarding the mechanisms of inactivation by pressure cycling and continuous HP treatments.Industrial relevanceThe resistance of bacterial spores to HP limits the industrial applications to refrigerated food products. In this study, we investigated the use of pressure cycling as a means to improve spore baroinactivation at moderate temperatures (T < 60 °C). We showed that cycling pressure does not significantly increase bacterial spore inactivation in comparable treatment duration, but certainly increases material fatigue in HP vessels. Thus, under moderate temperature, cycling pressure treatment is not industrially relevant.     

High-pressure destruction kinetics of Clostridium sporogenes spores in ground beef at elevated temperatures

High pressure (HP) is an alternative technique for thermal sterilization of foods with minimum quality loss. HP destruction kinetics of bacterial spores is essential to establishing sterilization process, but knowledge in this field is still very limited. In this study, destruction kinetics was investigated using Clostridium sporogenes PA 3679 (ATCC7955) spores in extra-lean ground beef (5 g each sealed in a sterile plastic bag). Duplicated samples were subjected to HP treatments at 700, 800 and 900 MPa in a HP system equipped with a Polyoxymethylene insulator to maintain constant temperatures at 80, 90 and 100 degrees C during pressure-holding time. The kinetic parameters of the spores (D- and Z-values) were evaluated at these pressures and temperatures. For the pressure from 700 to 900 MPa, D-values ranged from 15.8 to 7.0 and 1.5 to 0.63 min at 80 and 100 degrees C, respectively. The pressure resistance of Z(T)(P) value was 520-563 MPa at 80-100 degrees C. The temperature resistance of Z(P)(T) value was 19.1-19.7 degrees C at 700-900 MPa, much higher than that at atmospheric condition (12.4 degrees C). A regression model was generated which can be used to predict D-value or the death time of a minimum process under given pressure and temperature conditions. HP treatment with elevated temperatures can destroy bacterial spores with a shorter time or lower temperature than conventional thermal processing. This study provides useful information for the achievement of a safe HP sterilization process.     

Combined effects of high pressure,moderate heat and pH on the inactivation kinetics of Bacillus licheniformis spores in carrot juice

The objective of this research was to evaluate the combined effect of high pressure processing (temperature, pressure and time) and product (pH) related variables on destruction kinetics of spores of Bacillus licheniformis in carrot juice. A 3-level factorial experimental design was used with the microbial spores inoculated into carrot juice at the natural pH (6.2) and acidified pH (4.5 and 5.5), pressure (400, 500 or 600 MPa), temperature (40, 50, and 60 °C) and time (0–40 min) conditions. D values found varied from 0.6 to 14.1 min based on the temperature, pressure and pH level combinations. The corresponding temperature and pressure dependency of D values were in the range 23.3 to 31 °C and 241 to 465 MPa, respectively. The destruction pattern was also dependent on pH, with lower pH contributing to higher destruction rate. Conventional log-linear model and Weibull model were used to describe the survivor curves and for predicting processing time to achieve a 5D spore reduction. The survivor curves exhibited slightly upward concavity and therefore better described by a Weibull than the log-linear model. Treatment combinations showed significant (p ≤ 0.05) effects on D and z values of log-linear model and rate parameter (α) of Weibull model. The 5D spore count reduction times estimated using Weibull model parameters were longer than those from the log-linear model, generally demonstrating an over-treatment. Overall, the pH reduction of low acid foods showed a significant enhancement of rate of destruction of B. licheniformis spores.     

Kinetic studies on high-pressure inactivation of Bacillus stearothermophilus spores suspended in food matrices

Bacillus stearothermophilus spores ATCC 7953 can effectively be inactivated by high-pressure treatment, but only if it is applied at elevated temperatures; however, these temperatures are much lower compared to the temperature level used in heat inactivation under atmospheric pressure. Temperature and pressure in a range between 60 and 120°C and 50–600 MPa were applied to inactivate spores suspended in mashed broccoli and in cocoa mass. Utilizing an empirical mathematical model, derived from nth order kinetics, the survival curves of the spore strain investigated could be described accurately. The model can predict the impact of combined action of pressure and temperature on spore reduction. It was demonstrated that the inactivation of B. stearothermophilus spores ATCC 7953 improved with increasing treatment intensity. Beside intrinsic microbial inactivation mechanisms, the role of the pressure-induced shift in crystallization temperature of fat on spore inactivation in cocoa mass is discussed.     

Temperature uniformity mapping in a high pressure high temperature reactor using a temperature sensitive indicator

Recently, the first prototype ovomucoid-based pressure-temperature-time indicator (pTTI) for high pressure high temperature (HPHT) processing was described. However, for temperature uniformity mapping of high pressure (HP) vessels under HPHT sterilization conditions, this prototype needs to be optimized. To this end, this work aimed at the development of an ovomucoid-based indicator with combined pressure temperature dependent inactivation kinetics and a sufficient pressure temperature stability relevant for commercial HPHT sterilization. After varying buffer type and the pH at ambient pressure and temperature (pH_i), an indicator based on 1 g/L ovomucoid in 0.1 M MES-NaOH buffer pH_i 6.2 was selected. The inactivation behavior of this indicator system is characterized by pressure temperature dependent (combined Arrhenius-Eyring) first-order kinetics in the processing domain relevant for HPHT sterilization. This indicator showed good integrating properties under isobaric-isothermal and dynamic pressure temperature conditions.In a temperature uniformity study of a vertically oriented, pilot-scale HPHT vessel, pTTI readouts at different coordinates illustrated low and high temperature zones. As the inactivation of spores under HPHT is clearly positively temperature dependent, the food safety objective has to be verified in the former sampling zone.     

High Pressure Destruction Kinetics of Clostridium Sporogenes Spores in Salmon Slurry at Elevated Temperatures

Salmon slurry containing C. sporogenes spores was subjected to high pressure (HP) treatments (700–900 MPa; 80–100°C, and 0–24 min). Destruction rates (D value) and pressure/temperature sensitivity parameters (Z_P and Z_T ) were evaluated. Thermal treatment D values were an order of magnitude higher than those under HP. Higher pressures and temperatures accelerated the spore destruction rates. Z_P values were 14.5, 17.3 and 15.5°C at 700, 800 and 900 MPa respectively, while Z_T values (at constant temperature) were 440, 540, 550 MPa at 80, 90, and 100°C, respectively. The z value under thermal treatment was 8.8°C. The spores were relatively more sensitive to temperature than to pressure.     

Inhibition of nutrient- and high pressure-induced germination of Bacillus cereus spores by plant essential oils

The efficiency of high-pressure (HP) treatment to eliminate vegetative bacterial cells is synergistically increased by many natural antimicrobials, but the effects on spores are poorly described. Here we report the effect of eleven plant essential oils on the nutrient- and HP-induced germination of spores of a group VI psychrotolerant Bacillus cereus strain. Ten oils partially inhibited nutrient-induced germination. These oils also inhibited HP-induced germination, but some inhibited only germination at moderate (200 MPa) pressure and others only at very high (600 MPa) pressure. Inhibition of spore germination by essential oils may have an adverse effect on the effectiveness of spore inactivation by HP at moderate temperatures, and this should be taken into account when designing combined processes. Essential oil from carrot seed did not inhibit nutrient or HP germination although it showed growth inhibitory properties, and essential oils with these properties may therefore open interesting perspectives in combination treatments with HP.Industrial relevanceHP treatment is an alternative processing technique that preserves a better balance of food quality and microbiological safety as compared to thermal processing. While most vegetative bacteria are efficiently inactivated by HP, inactivation of spores is inefficient. At moderate temperature, spore inactivation proceeds in a two-step process in which spores first germinate and are subsequently inactivated. The combination with natural antimicrobials is a promising approach to enhance the efficiency of HP processing because it exerts a synergistic effect on inactivation of vegetative bacteria. However, the current work is one of the first to document the effect of essential oils on the HP-induced germination of spores.     

Accelerated inactivation of Geobacillus stearothermophilus spores by ohmic heating

Until recently, ohmic heating was commonly thought to kill microorganisms through a thermal effect. However a growing body of evidence suggests that non-thermal effects may occur. Our aim was to determine the kinetics of inactivation of Geobacillus stearothermophilus spores (ATCC 7953) under ohmic and conventional heating using a specially constructed test chamber with capillary sized cells to eliminate potential sources of error and ensure that identical thermal histories were experienced both by conventionally and ohmically heated samples. Ohmic treatments at frequencies of 60 Hz and 10 kHz were compared with conventional heating at 121, 125 and 130 °C for four different holding times. Both ohmic treatments showed a general trend of accelerated spore inactivation. It is hypothesized that vibration of polar dipicolinic acid molecules (DPA) and spore proteins to electric fields at high temperature conditions may result in the accelerated inactivation.     

The impact of high pressure and temperature on bacterial spores: inactivation mechanisms of Bacillus subtilis above 500 MPa

Abstract: High‐pressure thermal sterilization (HPTS) is an emerging technology to produce shelf stable low acid foods. Pressures below 300 MPa can induce spore germination by triggering germination receptors. Pressures above 500 MPa could directly induce a Ca⁺²‐dipicolinic acid (DPA) release, which triggers the cortex‐lytic enzymes (CLEs). It has been argued that the activated CLEs could be inactivated under HPTS conditions. To test this claim, a wild‐type strain and 2 strains of Bacillus subtilis spores lacking germinant receptors and one of 2 CLEs were treated simultaneously from 550 to 700 MPa and 37 to 80 C (slow compression) and at 60 to 80 C up to 1 GPa (fast compression). Besides, an additional heat treatment to determine the amount of germinated cells, we added TbCl₃ to detect the amount of DPA released from the spore core via fluorescent measurement. After pressure treatment for 120 min at 550 MPa and 37 °C, no inactivation was observed for the wild‐type strain. The amount of released DPA correlated to the amount of germinated spores, but always higher compared to the belonging cell count after pressure treatment. The release of DPA and the increase of heat‐sensitive spores confirm that the inactivation mechanism during HPTS passes through the physiological states: (1) dormancy, (2) activation, and (3) inactivation. As the intensity of treatment increased, inactivation of all spore strains also strongly increased (up to ?5.7 log₁₀), and we found only a slight increase in the inactivation of one of the CLE (sleB). Furthermore, above a certain threshold pressure, temperature became the dominant influence on germination rate. Practical Application: The continuous increase of high‐pressure (HP) research over the last several decades has already generated an impressive number of commercially available HP pasteurized products. Furthermore, research helped to provoke the certification of a pressure‐assisted thermal sterilization process by the U.S. FDA in February 2009. However, this promising sterilization technology has not yet been applied in industrial settings. An improved understanding of spore inactivation mechanisms and the ability to calculate desired inactivation levels will help to make this technology available for pilot studies and commercialization at an industrial scale. Moreover, if the synergy between pressure and elevated temperature on the inactivation rate could be identified, clarification of the underlying inactivation mechanism during HP thermal sterilization could help to further optimize the process of this emerging technology.     

Influence of thermosonication on Geobacillus stearothermophilus inactivation in skim milk

The influence of high intensity ultrasound coupled with thermoprocessing on the inactivation of Geobacillus stearothermophilus vegetative cells and spores in skim milk powder was explored using response surface methodology and two polynomial models were developed. Optimization of cell reduction (4.8 log) was found to be at 19.75% total solids (TS), 45 °C, and 30 s, while optimization of spore reduction (0.45 log) was found to be at 31.5% TS, 67.5 °C, and 17.5 s. Model verification experiments were performed using common milk powder processing conditions. Results showed the inactivation of cells and spores to be most effective before (9.2% TS, 75 °C, and 10 s) and after (50% TS, 60 °C, and 10 s) the evaporator during milk powder processing and may produce an additive effect in microbial reduction when the two locations are combined, resulting in a 5.8 log reduction for vegetative cells and 0.51 log reduction for spores.     

Hydrophobic properties and extraction of Bacillus anthracis spores from liquid foods

The objectives of this study were to characterize the hydrophobic properties of three strains of Bacillus anthracis using the microbial adherence to hydrocarbons (MATH) assay and determine the recovery of spores by hexadecane extraction from water, milk and orange juice using a modified version of this assay. In water mixtures, the hydrophobicity of B. anthracis spores ranged from 5 to 80% as the concentration of hexadecane and the mixing time increased. Two of the three strains showed significantly different hydrophobicity values. Increased pre-incubation temperature of the spore suspension had inconsistent effects on hydrophobicity across the three strains. The hydrophobicity of spores did not change significantly during storage at 4 °C. However, recovery of spores in the hexadecane fraction from aqueous mixtures was always less than 5% even at conditions in which the hydrophobicity values were greater than 40%. The recovery of spores in the hexadecane fraction increased to almost 20% when the hexadecane was mixed with milk or orange juice, although the majority of spores remained in the aqueous phase. The B. anthracis spores were relatively hydrophobic according to the MATH assay, but this test was not a good predictor of the partitioning of B. anthracis spores to hexadecane. The separation of B. anthracis from food matrices using hexadecane extraction was ineffective. Although the modified MATH assay was not able to efficiently extract B. anthracis from various food media, development of methods for rapid concentration and separation of this and other select agents from food remains vital to food defense.     

REMOVAL KINETICS OF BACILLUS CEREUS SPORES FROM STAINLESS STEEL PIPES UNDER CIP PROCEDURE: INFLUENCE OF SOILING AND CLEANING CONDITIONS

Cleaning efficiency is of prime importance for food industries to ensure both the quality and safety of the products. The removal kinetics of Bacillus cereus spores adhering to unheated stainless steel pipes was studied under turbulent flow conditions (Reynolds’number of 77500 and 116300) in order to be close to those encountered in industrial practice. The experimental data was fitted using a hyperbolic tangent model. Variance analysis was then performed to underline any potential effects on the kinetics of the processing parameters, such as soiling conditions, soiling media and mean walls shear stress during cleaning. A significant influence of the adhesion medium (milk or saline) is shown at the level of spore removal (P < 0.001). This trend could probably be explained by the change in the surface properties of spores and stainless steel surfaces when covered by milk macromolecules. After milk soiling in turbulent flow conditions, removal efficiency was enhanced by a factor of 2.5 to remove 50% of the initial spore contamination and by a factor of 2 for the remaining spores after 30 min of cleaning. No effect of the two mean wall shear stresses (9.4 and 19.1 Pa) has been identified. The removal kinetic model proposed here could now permit the effect on the cleaning efficiency of a wide range of CIP conditions to be tested.     

EVALUATION OF A CHEMICAL MARKER FOR PROCESS LETHALITY MEASUREMENT AT HOC IN A CONTINUOUS FLOW HOLDING TUBE

Meat beads containing glucose precursor of a chemical marker and alginate beads containing spores of Bacillus stearothermophilus were prepared and subjected to steam heating at HOC for selected time intervals. Marker yields were related to spore survivor data and lethality values obtained from time-temperature data to generate calibration curves. Meat balls, fabricated with meat (marker precursor) and alginate (microbial spores) beads placed at the center, were heated at 110 ± 0.5C for selected time intervals (0–55 min) in a continuous flow holding tube. During the heat treatment, the panicles were held stationary in the holding tube while the carrier fluid (0.5% CMC solution) was circulated at 2.6 gallons per min. Transient time-temperature responses of the particles were recorded at bead locations during test runs. All treated samples were analyzed for marker yield as well as spore survivors. Using the calibration curves and marker yield data under test conditions, the corresponding spore count reduction and accumulated lethalities were computed. Lethalities and spore count reductions calculated from marker yield data showed good correlations with those obtained from the experiment. The results indicate that the chemical marker has good potential to provide data on accumulated lethality and spore count reductions. Prior to extension of this approach to aseptic processing conditions, additional kinetic data on spore destruction and marker formation should be gathered at temperatures applicable to these processes.     

Estimation of Accumulated Lethality Under Pressure-Assisted Thermal Processing

A study was conducted to develop an integrated process lethality model for pressure-assisted thermal processing (PATP) taking into consideration the lethal contribution of both pressure and heat on spore inactivation. Assuming that the momentary inactivation rate was dependent on the survival ratio and momentary pressure–thermal history, a differential equation was formulated and numerically solved using the Runge–Kutta method. Published data on combined pressure–heat inactivation of Bacillus amyloliquefaciens spores were used to obtain model kinetic parameters that considered both pressure and thermal effects. The model was experimentally validated under several process scenarios using a pilot-scale high-pressure food processor. Using first-order kinetics in the model resulted in the overestimation of log reduction compared to the experimental values. When the n ^th-order kinetics was used, the computed accumulated lethality and the log reduction values were found to be in reasonable agreement with the experimental data. Within the experimental conditions studied, spatial variation in process temperature resulted up to 3.5 log variation in survivors between the top and bottom of the carrier basket. The predicted log reduction of B. amyloliquefaciens spores in deionized water and carrot purée had satisfactory accuracy (1.07–1.12) and regression coefficients (0.83–0.92). The model was also able to predict log reductions obtained during a double-pulse treatment conducted using a pilot-scale high-pressure processor. The developed model can be a useful tool to examine the effect of combined pressure–thermal treatment on bacterial spore lethality and assess PATP microbial safety.     

INACTIVATION KINETICS AND REDUCTION OF BACILLUS COAGULANS SPORE BY THE COMBINATION OF HIGH PRESSURE AND MODERATE HEAT

The combination effect of high pressure (400, 500 and 600 MPa) and moderate heat (70 and 80C) on the inactivation kinetics and reduction of Bacillus coagulans spore in phosphate buffer and ultra-high temperature (UHT) whole milk was investigated. The pressure come-up time and corresponding logarithmic reduction of spore inactivation were considered during pressure-thermal treatment. B. coagulans spore had a much higher resistance to pressure in UHT whole milk than in phosphate buffer. Survival data were modeled using the linear, Weibull and log-logistic models to obtain relevant kinetic parameters. The tailing phenomenon occurred in all survival curves, indicating the linear model was not adequate for describing these curves. The mean square error and regression coefficient suggested that the log-logistic model produced best fits to all survival curves, followed by the Weibull model.

PRACTICAL APPLICATIONS

It becomes increasingly apparent that high-pressure treatment combined with moderate heat treatment for low acid and acid products is often required for effective bacterial spores' inactivation. Consequently, the prediction model of microbial survival curves is essential. Bacillus coagulans is a slightly pressure-resistant and relatively heat-resistant spoilage bacterium of considerable concern during the processing of acid foods. Spore inactivation effect during the pressure come-up time is sometimes considerable and should not beignored. The use of mathematical models to predict inactivation for spores could help the food industry further to develop optimum process conditions.