Predicting thermal inactivation in media of different pH of Salmonella grown at different temperatures

The influence of the growth temperature and the pH of the heating medium on the heat resistance at different temperatures of Salmonella typhimurium ATCC 13311 was studied and described mathematically. The shift of the growth temperature from 10 to 37 degrees C increased heat resistance of S. typhimurium fourfold. The pH of the heating medium at which heat resistance was maximum was pH 6 for cells grown at 37 degrees C, but changed with growth temperature. The alkalinization of the heating medium from pH 6 to pH 7.7 decreased the heat resistance of cells grown at 37 degrees C by a factor of 3. Neither the growth temperature nor the pH modified the z values significantly (4.9 degrees C). The decimal reduction times at different treatment temperatures, in buffers of different pH of cells of S. typhimurium grown at different temperatures, were accurately described by a mathematical equation (correlation coefficient of 0.97). This equation was also tested for Salmonella senftenberg 775W (ATCC 43845) and Salmonella enteritidis ATCC 13076, strains in which the correlation coefficients between the observed and the theoretically calculated values were 0.91 and 0.98, respectively.     

Relationship between the apparent heat resistance of Bacillus cereus spores and the pH and NaCl concentration of the recovery medium

Conventional heat resistance data, D values, were previously established by other workers at optimal condition for spores outgrowth. However, in canned food conditions of outgrowth are generally suboptimal in term of pH, salt concentration, water activity. The combined effects of pH and NaCl level of the recovery medium for the D value and z(pH) value were studied. Spores of Bacillus cereus were heated at 95 degrees C in phosphate-citrate buffer media at pH 7. Cells were recovered at 25 degrees C in Nutrient Agar with pH ranging from 5 to 7 and 1% to 4% (w/w) NaCl concentration. For each condition D' values (decimal reduction time associated with the recovery media characteristics) were determined. The results show a major influence of the recovery pH on the D' values. This effect is characterised by the z'(pH) values, distance of recovery medium pH from optimum recovery pH* medium (6.7) which leads to a tenfold reduction time of D value. The increase of the salt concentration leads to a slight decrease of D' value. However z'(pH) values are not significantly affected by the salt concentration. A simple three parameter model describing the effects of pH and NaCl concentration of the recovery medium upon the heat resistance of spores is proposed. The interaction between pH and salt concentration is sufficiently low to be neglected by the model.     

Quantifying the combined effects of the heating time, the temperature and the recovery medium pH on the regrowth lag time of Bacillus cereus spores after a heat treatment

The purpose of this study was to quantify the lag time of re-growth of heated spores of Bacillus cereus as a function of the conditions of the heat treatment: temperature, duration and pH of the recovery medium. For a given heating temperature, curves plotting lag times versus time of heating show more or less complex patterns. However, under a heating time corresponding to a decrease of 2 decimal logarithms of the surviving populations of spores, a linear relationship between the lag time of growth and the time of the previous heat treatment can be observed. The slope of this linear relationship followed itself a Bigelow type linear relationship, the slope of which yielded a zeta-value very close to the observed conventional z-value. It was then concluded that the slope of the regrowth lag time versus the heating time followed a linear relationship with the sterilisation value reached in the course of the previous heat treatment. A sharp effect of the pH of the medium which could be described by a simple "secondary" model was observed. As expected, the observed intercept of the linear relationship between lag time and heating time (lag without previous heating) was dependent on only the pH of the medium and not on the heating temperature.     

Heat resistance of Bacillus cereus, Salmonella typhimurium and Lactobacillus delbrueckii in relation to pH and ethanol

The aim of the present work was to investigate the effect of ethanol alone and in combination with low pH on the heat resistance of specific bacteria. The bacteria chosen are representative of heat resistant and heat sensitive pathogens (Bacillus cereus and Salmonella typhimurium) and of relatively heat resistant spoilage microorganisms (Lactobacillus delbrueckii). The chosen bacteria were treated at different temperatures ranging between 70 and 97 degrees C for B. cereus, 48 and 54 degrees C for S. typhimurium and 44 and 60 degrees C for L. delbrueckii, in media of different pH (3, 5 and 7) and ethanol content (0, 2.5, 5, 7.5 and 10%). Both factors proved to be very effective in reducing the heat resistance of the bacteria examined in this work. At pH 7, an increase in ethanol content up to 10% caused D values to decrease by up to 100-fold (S. typhimurium). A drop from pH 7 to pH 3 also caused up to a 100-fold reduction in the D values (S. typhimurium). For B. cereus the regression analysis of the log10 of the D value in relation to temperature, pH and ethanol content was used to produce a second order polynomial equation. The z values increased at decreasing pH and cells were more sensitive to ethanol at lower pH. For S. typhimurium the polynomial equation produced to describe the relationship between log10 of the D values and temperature and ethanol content was also a second order equation indicating that the relationship between z values and ethanol was non-linear. For L. delbrueckii, z values were independent of the ethanol content of the heating medium. Acid tolerance at 25 and 37 degrees C (L. delbrueckii and S. typhimurium) and acid adaptation (S. typhimurium) were also tested. L. delbrueckii was more ethanol and pH tolerant than S. typhimurium at heat treatment temperatures whilst S. typhimurium was more acid tolerant than L. delbrueckii at incubation temperatures (25 and 37 degrees C). Acid adaptation increased the tolerance of S. typhimurium to low pH at 25 degrees C but failed to improve its thermal resistance at 48 degrees C. In all bacteria used, the effects of pH and ethanol were more evident at lower treatment temperatures and therefore their significance becomes greater in view of reduced thermal processing and/or changes in product formulation.     

Survival curves of heated bacterial spores: effect of environmental factors on Weibull parameters

The classical D-value of first order inactivation kinetic is not suitable for quantifying bacterial heat resistance for non-log linear survival curves. One simple model derived from the Weibull cumulative function describes non-log linear kinetics of micro-organisms. The influences of environmental factors on Weibull model parameters, shape parameter "p" and scale parameter "delta", were studied. This paper points out structural correlation between these two parameters. The environmental heating and recovery conditions do not present clear and regular influence on the shape the parameter "p" and could not be described by any model tried. Conversely, the scale parameter "delta" depends on heating temperature and heating and recovery medium pH. The models established to quantify these influences on the classical "D" values could be applied to this parameter "delta". The slight influence of the shape parameter p variation on the goodness of fit of these models can be neglected and the simplified Weibull model with a constant p-value for given microbial population can be applied for canning process calculations.     

Modelling the influence of palmitic, palmitoleic, stearic and oleic acids on apparent heat resistance of spores of Bacillus cereus NTCC 11145 and Clostridium sporogenes Pasteur 79.3

Heat resistance of spores is affected by many factors such as temperature, pH, water activity (aw) and others. Previous studies have reported that free fatty acids can affect the germination and growth of bacterial spores. In this study, we investigated the influence of free fatty acids in heating medium or in recovery medium on the heat resistance of spores of Bacillus cereus NTCC 11145 and Clostridium sporogenes Pasteur 79.3. Four free fatty acids were studied: palmitic, palmitoleic, stearic and oleic acids. During thermal treatments, the impact of these FFA in heating media was generally low, but the presence of free fatty acids in the recovery medium highly decreases bacterial spore apparent heat resistance, particularly with unsaturated fatty acids. A mathematical model was developed to describe and quantify the influence of free fatty acids in recovery media on the D-values. The z′_FFA parameter values which quantify the impact of free fatty acids were determined. The variation of this parameter value according to the free fatty acid type was compared with MIC value variation given in the literature. The model enables the decrease in D-values in the presence of free fatty acids to be estimated. The high concentrations of free fatty acids in liver or canned duck may explain the microbial stability with low sterilization values applied.     

Modelling the influence of pH and organic acid types on thermal inactivation of Bacillus cereus spores

A model is proposed to describe the influence pH on the heat resistance of Bacillus cereus spores. In addition to the conventional z value, the effect of pH on the thermal resistance of spores is characterised by a z(pH) value (z(pH) is the distance of pH from a reference pH*, which leads to a 10-fold reduction of D value). The type of organic acid used for acidifying the heating medium, influences the z(pH) value. For nine organic acids, a linear relationship between the calculated z(pH) value and its lower acid pKa is observed. This relationship showed that the acid form (dissociated or undissociated) modifies the thermal spore resistance in addition to the H+ ion. The influence of acetic acid concentration on the D value at pH 7 shows the protective effect of the dissociated acid form on the heat resistance of spores. The acid concentration in the medium modified the heat resistance of spore and the z(pH) value.     

Effect of the medium characteristics and the heating and cooling rates on the nonisothermal heat resistance of Bacillus sporothermodurans IC4 spores

In recent years, highly thermo-resistant mesophilic spore-forming bacteria belonging to the species Bacillus sporothermodurans have caused non-sterility problems in industrial sterilization processes. The aim of this research was to evaluate the effect of the heating medium characteristics (pH and buffer/food) on the thermal inactivation of B. sporothermodurans spores when exposed to isothermal and non-isothermal heating and cooling treatments and the suitability of non-linear Weibull and Geeraaerd models to predict the survivors of these thermal treatments. Thermal treatments were carried out in pH 3, 5 and 7 McIlvaine buffer and in a courgette soup. Isothermal survival curves showed shoulders that were accurately characterized by means of both models. A clear effect of the pH of the heating medium was observed, decreasing the D₁₂₀ value from pH 7 to pH 3 buffer down to one third. Differences in heat resistance were similar, regardless of the model used and were kept at all temperatures tested. The heat resistance in courgette soup was similar to that shown in pH 7 buffer. When the heat resistance values obtained under isothermal conditions were used to predict the survival in the non-isothermical experiments, the predictions estimated the experimental data quite accurately, both with Weibull and Geeraerd models.     

Predicting heat inactivation of Staphylococcus aureus under nonisothermal treatments at different pH

The aim was to assess whether heat resistance data obtained from isothermal treatments allow the estimation of survivors of Staphylococcus aureus under nonisothermal conditions and to find a model that accurately predicts its heat inactivation at constantly rising heating rates (0.5-9 degrees C/min) in media of different pH (4.0-7.4). S. aureus showed a higher heat resistance under isothermal treatments at pH 4.0 than at pH 5.5-7.4. However, under nonisothermal treatments S. aureus increased its heat resistance at pH 5.5-7.4 and became more thermotolerant than at pH 4.0. Estimations of survival curves under nonisothermal treatments obtained from heat resistance parameters of isothermal treatments did not adequately fit experimental values. Whereas the number of survivors was much higher than estimated at pH 5.5-7.4, that obtained at the slower heating rates at pH 4.0 was lower. An equation based on the Weibullian-like distribution (log10 S(t) = (t/delta)p) accurately described survival curves obtained under nonisothermal conditions. A nonlinear relationship was observed among the scale parameter (delta) and the heating rate which allowed the development of two equations capable of predicting the inactivation rate of S. aureus under nonisothermal treatments. This study might contribute to prevent public health risks in foods requiring long heating lag phases during their processing.     

Effect of sodium chloride concentration on the heat resistance and recovery of Salmonella typhimurium

The survival of Salmonella typhimurium (ATCC 13311) heated and recovered in media with 0, 1, 2, 3, 4 or 5% (w/w) added sodium chloride was investigated. A protective effect in the heating medium and an inhibitory effect in the recovery medium were observed. The results showed an interaction between the effect on, D(58 degrees C) values, of sodium chloride concentration in both media. Lower concentration in the heating media led to a greater effect of the sodium chloride concentration in the recovery media. When the sodium chloride concentration was the same in both media, the protective effect exerted in the heating media dominated over its inhibitory effect in the recovery media.     

不同介质热处理对油脂酸值和羰基值的影响

根据正交实验的设计理论,以加热时间、加热温度、食盐加入量、味精加入量为油脂品质的影响因素设计正交实验,分别以铁锅和不粘锅为加热介质对大豆油进行不同环境下的热处理,测定处理后油脂的酸值及羰基值,比较几种因素对油脂品质的影响。结果表明,加热时间对油脂酸值及羰基值的影响最大,其次是加热温度,食盐及味精加入量对酸值和羰基值的影响最小,不粘锅环境下油脂的酸值和羰基值整体比铁锅环境下大,表明加热介质对油脂品质有影响,铁锅环境下油脂品质变化相对较小。     

Modeling heat resistance of Bacillus weihenstephanensis and Bacillus licheniformis spores as function of sporulation temperature and pH

Although sporulation environmental factors are known to impact on Bacillus spore heat resistance, they are not integrated into predictive models used to calculate the efficiency of heating processes. This work reports the influence of temperature and pH encountered during sporulation on heat resistance of Bacillus weihenstephanensis KBAB4 and Bacillus licheniformis AD978 spores. A decrease in heat resistance (δ) was observed for spores produced either at low temperature, at high temperature or at acidic pH. Sporulation temperature and pH maximizing the spore heat resistance were identified. Heat sensitivity (z) was not modified whatever the sporulation environmental factors were. A resistance secondary model inspired by the Rosso model was proposed. Sporulation temperatures and pHs minimizing or maximizing the spore heat resistance (T(min(R)), T(opt(R)), T(max(R)), pH(min(R)) and pH(opt(R))) were estimated. The goodness of the model fit was assessed for both studied strains and literature data. The estimation of the sporulation temperature and pH maximizing the spore heat resistance is of great interest to produce spores assessing the spore inactivation in the heating processes applied by the food industry.     

Predictive thermal inactivation model for Listeria monocytogenes with temperature, pH, NaCl, and sodium pyrophosphate as controlling factors.

The effects and interactions of heating temperature (55 to 65 degrees C), pH (4 to 8), salt (NaCl; 0 to 6%, wt/vol), and sodium pyrophosphate (SPP; 0 to 0.3%, wt/vol) on the heat inactivation of a four-strain mixture of Listeria monocytogenes in beef gravy were examined. A factorial experimental design comparing 48 combinations of heating temperature, salt concentration, pH value, and SPP content was used. Heating was carried out using a submerged-coil heating apparatus. The recovery medium was plate count agar supplemented with 0.6% yeast extract and 1% sodium pyruvate. Decimal reduction times (D-values) were calculated by fitting a survival model to the data with a curve-fitting program. The D-values were analyzed by second-order response surface regression for temperature, pH, NaCl, and SPP levels. Whereas increasing the NaCl concentration protected L. monocytogenes against the lethal effect of heat, high SPP concentrations increased heat sensitivity. Also, low pH values increased heat sensitivity of L. monocytogenes. The four variables interacted to affect the inactivation of the pathogen. Thermal resistance of L. monocytogenes can be lowered by combining these intrinsic factors. A predictive model that described the combined effect of temperature, pH, NaCl, and SPP levels on thermal resistance of L. monocytogenes was developed. The model can predict D-values for any combination of temperature, pH, NaCl, and SPP that are within the range of those tested. Using this predictive model, food processors should be able to design adequate thermal regimes to eliminate L. monocytogenes in thermally processed foods.     

Influence of pH on heat resistance of spores of Bacillus coagulans in buffer and homogenized foods

The influence of pH of heating menstruum (McIlvaine buffer) on the heat resistance of Bacillus coagulans spores has been investigated and compared with the heat resistance in homogenized tomato and asparagus at pH 7 and 4 at a wide range of temperatures. Spores were less heat resistant in all menstrua at acid pH. The magnitude of this effect was greatest at the lowest heating temperatures tested. z values in buffer increased from 8.9 degrees C at pH 7 to 10.5 degrees C at pH 4. pH of menstrua was the main influencing factor, but media composition also influenced heat resistance: at pH 7 heat resistance was similar in all menstrua (D111 degrees C = 1.6 min) but at pH 4 the heat resistance in homogenized foods (D111 degrees C = 0.26 min in tomato and D111 degrees C = 0.28 min in asparagus) was lower than in buffer (D111 degrees C = 0.49 min). The reduced influence of the acidification of media on the heat resistance of B. coagulans at higher temperatures should be taken into account when a rise in the temperature of treatment for canned vegetables is considered to shorten duration of heat processes.     

Effect of pH at heating on the acid-induced aggregation of casein micelles in reconstituted skim milk

Reconstituted skim milk samples at pH between 6.5 and 7.1 (heating pH) were heated at 80°C, 90°C or 100°C for 30 min (heating temperature). The particle size of the casein micelles was measured at pH 4.75-7.1 (measurement pH) and at temperatures of 10°C, 20°C and 30°C (measurement temperature) using photon correlation spectroscopy. The particle size of the casein micelles, at a measurement pH of 6.7 and a measurement temperature of 20°C, was dependent on the heating pH and heating temperature to which the milk was subjected. The casein micelle size in unheated milk was about 215 nm. At a heating pH of 6.5, the casein micelle size increased by about 15, 30 and 40 nm when the milk was heated at 80°C, 90°C or 100°C, respectively. As the heating pH of the milk was increased, the size of the casein micelles decreased so that, at pH 7.1, the casein micelles were ∼20 nm smaller than those from unheated milk. Larger effects were observed as the heating temperature was increased from 80°C to 100°C. The size differences as a consequence of the heating pH were maintained at all measurement temperatures and at all measurement pH down to the pH at which aggregation of the micelles was observed. For all samples, size measurements at 10°C showed no aggregation at all measurement pH. Aggregation occurred at progressively higher pH as the measurement temperature was increased. Aggregation also occurred at a progressively higher measurement pH as the heating pH was increased. The particle size changes on heating and the aggregation on subsequent acidification may be related to the pH dependence of the association of whey proteins with, and the dissociation of κ-casein from the casein micelles as milk is heated.     

Responses of Listeria monocytogenes to acid stress and glucose availability monitored by measurements of intracellular pH and viable counts

Physiological aspects of the response of Listeria monocytogenes to acidic conditions and effect of glucose availability were studied by fluorescence ratio-imaging microscopy (FRIM) as compared with traditional viable counts. Three types of experiments were conducted: (i) static with measurements of intracellular pH (pHi) at extracellular pH (pHo) values ranging from pH 3.0 to 6.0 at 0.5 pH unit intervals; (ii) kinetic with monitoring of bacterial responses to changes in the pHo from the value of 6.0 to 4.0 or 3.0; (iii) survival experiments studying bacterial recovery in response to a shift to favourable conditions after a treatment at low pH. All the experiments were performed at three levels of glucose in the medium (0, 1, and 10 mM). Both survival and pHi were greatly affected by pHo and glucose availability with the highest values for CFU and pHi at highest glucose concentration and pHo values in the medium in all trials. A high correlation (R2 = 0.995) between pHi and CFU counts was observed. The pH gradient started to collapse at pHo 4 and below for trials with glucose in the medium and at pHo 5.5 and below without glucose. A recovery step was proposed after the apparently lethal treatment to assess cell viability by FRIM.     

Heat resistance of Yersinia enterocolitica grown at different temperatures and heated in different media.

In the range of 4-20 degrees C, growth temperature did not influence the heat resistance at 54-66 degrees C for Yersinia enterocolitica at pH 7 in citrate phosphate buffer. However, when cells were grown at 37 degrees C. the D62 increased from 0.044 to 0.17 min. This increase was constant at all heating temperatures tested (z = 5.7-5.8). Growth temperature did not influence the proportion of heat-damaged cells after a heat treatment, as measured by their response to a 2% of sodium chloride added to the recovery medium. The sensitivity of heat treated cells to nisin or lysozyme depended on growth temperature: Whereas the number of cells grown at 4 degrees C surviving heat treatment was the same regardless of the presence of 100 IU/ml of nisin or 100 microg/ml of lysozyme in the recovery medium, that of cells grown at 37 degrees C was, in these media, lower. The pH of maximum heat resistance in citrate phosphate buffer was pH 7 for cells grown at 37 degrees C, but pH 5 for those grown at 4 degrees C. In both suspensions the magnitude of the effect of pH on heat resistance was constant at all heating temperatures. For cells grown at 4 degrees C the heat resistance at 54-66 degrees C, in skimmed milk or pH 7 buffer, was the same. For cells grown at 37 degrees C this also applied for heat treatment at 66 degrees C but at 56 degrees C the heat resistance in skimmed milk was higher.     

Validation of an overall model describing the effect of three environmental factors on the apparent D-value of Bacillus cereus spores

Several factorial models extending the famous Bigelow model to describe the influence of the heating and recovery pH and a_w conditions on bacterial heat resistance have been developed. These models can be associated in an overall multifactorial model describing the influences of heating and recovery conditions on D values. For Bacillus cereus strain ADQP 407 the model parameters characterising the environmental factor influences (pH, Temperature, a_w) were evaluated. Determination of bacterial heat resistance in cream chocolate have been realised to validate these parameter values and to evaluate the level of the influence of food texture or different compounds not taken account of in the model.     

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.     

Predicting heat inactivation of Listeria monocytogenes under nonisothermal treatments

The aim of this study was to find a model that accurately predicts the heat inactivation of Listeria monocytogenes (ATCC 15313) at constantly rising heating rates (0.5 to 9 degrees C/min) in media of different pH values (4.0 to 7.4). Survival curves of L. monocytogenes obtained under isothermal treatments at any temperature were nearly linear. Estimations of survival curves under nonisothermal treatments obtained from heat resistance parameters of isothermal treatments adequately fit experimental values obtained at pH 4.0. On the contrary, survivors were much higher than estimations at pH 5.5 and 7.4. The slower the heating rate and the longer the treatment time, the greater the differences between the experimental and estimated values. An equation based on the Weibullian-like distribution, log S(t) = (t/delta)p, accurately described survival curves of L. monocytogenes obtained under nonisothermal conditions within the range of heating rates investigated. A nonlinear relationship was observed between the scale parameter (delta) and the heating rate, which allowed the development of an equation capable of predicting the inactivation rate of L. monocytogenes under nonisothermal treatments at pH 5.5 and 7.4. The model predictions were a good fit to the measured data independent of the magnitude of the thermotolerance increase. This work might contribute to the increase in safety of those food products that require long heating lag phases during the pasteurization process.