Predictive model for growth of Clostridium botulinum from spores at temperatures applicable to cooling of cooked ground pork

Cooling deviations and temperature abuse are two main reasons leading to the risk of Clostridium botulinum outgrowth in cooked pork. The aim of this research was to create a model that could be used to estimate C. botulinum growth from spores in cooked pork at temperatures similar to those used to chill cooked pork in processing facilities and food establishments. A cocktail of proteolytic C. botulinum types A and B consisting of five strains per type were used to inoculate pork to a final spore concentration of approximately 2 log CFU/g and cooked to 71 °C to heat shock the spores and kill vegetative microbes. The growth of C. botulinum was established at constant storage temperatures from 10 to 46 °C. C. botulinum growth was also studied under dynamic temperature conditions with cooling set to start at 54.4 °C and end at 4.4 °C or 7.2 °C in monophasic or biphasic cooling profiles, respectively. Growth parameters were estimated using the Baranyi model as a primary model and growth rates were fitted using the modified Ratkowsky secondary model with respect to temperature. The R² values ranged from 0.7653 to 0.9995 indicating that the Baranyi primary model was well suited to the growth data. The modified Ratkowsky secondary model's R² was 0.9653 and its root mean square error (RMSE) was 0.0687. All 11 prediction error values computed were within the limit of acceptable prediction zone (−1.0 to 0.5) suggesting a good fit of the model. The predictive model can provide information for the safety of cooked pork exposed to longer chilling times or for customized process schedule development as cooling of larger diameter products presents a processing challenge in the meat process operations.     

Predictive model for growth of Clostridium perfringens during cooling of cooked uncured meat and poultry

Comparison of Clostridium perfringens spore germination and outgrowth in cooked uncured products during cooling for different meat species is presented. Cooked, uncured product was inoculated with C. perfringens spores and vacuum packaged. For the isothermal experiments, all samples were incubated in a water bath stabilized at selected temperatures between 10 and 51 °C and sampled periodically. For dynamic experiments, the samples were cooled from 54.4 to 27 °C and subsequently from 27 to 4 °C for different time periods, designated as x and y hours, respectively. The growth models used were based on a model developed by Baranyi and Roberts (1994. A dynamic approach to predicting bacterial growth in food. Int. J. Food Micro. 23, 277-294), which incorporates a constant, referred to as the physiological state constant, q₀. The value of this constant captures the cells’ history before the cooling begins. To estimate specific growth rates, data from isothermal experiments were used, from which a secondary model was developed, based on a form of Ratkowsky’s 4-parameter equation. The estimated growth kinetics associated with pork and chicken were similar, but growth appeared to be slightly greater in beef; for beef, the maximum specific growth rates estimated from the Ratkowsky curve was about 2.7 log₁₀ cfu/h, while for the other two species, chicken and pork, the estimate was about 2.2 log₁₀ cfu/h. Physiological state constants were estimated by minimizing the mean square error of predictions of the log₁₀ of the relative increase versus the corresponding observed quantities for the dynamic experiments: for beef the estimate was 0.007, while those for pork and chicken the estimates were about 0.014 and 0.011, respectively. For a hypothetical 1.5 h cooling from 54 °C to 27° and 5 h to 4 °C, corresponding to USDA-FSIS cooling compliance guidelines, the predicted growth (log₁₀ of the relative increase) for each species was: 1.29 for beef; 1.07 for chicken and 0.95 log₁₀ for pork. However, it was noticed that for pork in particular, the model using the derived q₀ had a tendency to over-predict relative growth when the observed amount of relative growth was small, and under-predict the relative growth when the observed amount of relative growth was large. To provide more fail-safe estimate, rather than using the derived value of q₀, a value of 0.04 is recommended for pork.     

Dynamic predictive model for the growth of Salmonella spp. in liquid whole egg

Abstract: A dynamic model for the growth of Salmonella spp. in liquid whole egg (LWE) (approximately pH 7.8) under continuously varying temperature was developed. The model was validated using 2 (5 to 15 °C; 600 h and 10 to 40 °C; 52 h) sinusoidal, continuously varying temperature profiles. LWE adjusted to pH 7.8 was inoculated with approximately 2.5–3.0 log CFU/mL of Salmonella spp., and the growth data at several isothermal conditions (5, 7, 10, 15, 20, 25, 30, 35, 37, 39, 41, 43, 45, and 47 °C) was collected. A primary model (Baranyi model) was fitted for each temperature growth data and corresponding maximum growth rates were estimated. Pseudo‐R² values were greater than 0.97 for primary models. Modified Ratkowsky model was used to fit the secondary model. The pseudo‐R² and root mean square error were 0.99 and 0.06 log CFU/mL, respectively, for the secondary model. A dynamic model for the prediction of Salmonella spp. growth under varying temperature conditions was developed using 4th‐order Runge–Kutta method. The developed dynamic model was validated for 2 sinusoidal temperature profiles, 5 to 15 °C (for 600 h) and 10 to 40 °C (for 52 h) with corresponding root mean squared error values of 0.28 and 0.23 log CFU/mL, respectively, between predicted and observed Salmonella spp. populations. The developed dynamic model can be used to predict the growth of Salmonella spp. in LWE under varying temperature conditions. Practical Application: Liquid egg and egg products are widely used in food processing and in restaurant operations. These products can be contaminated with Salmonella spp. during breaking and other unit operations during processing. The raw, liquid egg products are stored under refrigeration prior to pasteurization. However, process deviations can occur such as refrigeration failure, leading to temperature fluctuations above the required temperatures as specified in the critical limits within hazard analysis and critical control point plans for the operations. The processors are required to evaluate the potential growth of Salmonella spp. in such products before the product can be used, or further processed. Dynamic predictive models are excellent tools for regulators as well as the processing plant personnel to evaluate the microbiological safety of the product under such conditions.     

Predictive model for growth of Clostridium perfringens during cooling of cooked ground pork

A predictive dynamic model for Clostridium perfringens spore germination and outgrowth in cooked pork products during cooling is presented. Cooked, ground pork was inoculated with C. perfringens spores and vacuum packaged. For the isothermal experiments, all samples were incubated in a water bath stabilized at selected temperatures between 10 and 51 °C and sampled periodically. For dynamic experiments, the samples were cooled from 54.4 to 27 °C and subsequently from 27 to 4 °C for different time periods, designated as x and y hours, respectively. The growth models used were based on a model developed by Baranyi and Roberts (1994), which incorporates a constant, referred to as the physiological state constant, q₀. The value of this constant captures the cells' history before the cooling begins. To estimate specific growth rates, data from isothermal experiments were used, from which a secondary model was developed, based on a particular form of Ratkowsky's 4-parameter equation. Using the data from dynamic experiments and the Ratkowsky model, an optimal value of q₀ (=0.01375) was derived minimizing the mean square error of predictions. However, using this estimate, the model had a tendency to over-predict relative growth when there was observed small amounts of relative growth, and under-predict relative growth when there was observed large relative growth. To provide more fail-safe estimates, rather than using the derived value of q₀, a value of 0.04 is recommended. The predictive model with this value of q₀ would provide more fail-safe estimates of relative growth and could aid producers and regulatory agencies with determining disposition of products that were subjected to cooling deviations.Industrial relevanceSafe time/temperature for cooling of cooked pork is very important to guard against the pathogen in cooked products. Predictive model will assist industry to determine compliance with regulatory performance standards and to ensure microbiological safety of cooked products.     

Analysis of mathematical models of Pseudomonas spp. growth in pallet-package pork stored at different temperatures

Pseudomonas of pallet-packaged raw pork grown at 0, 5, 10, 15, 20 and 25 °C has been studied in this paper. The modified Gompertz, Baranyi and Huang models were used for data fitting. Statistical criteria such as residual sum of squares, mean square error, Akaike's information criterion, and pseudo-R² were used to evaluate model performance. Results showed that there was an apparent decline in Pseudomonas growth at initial-storage phase at low temperatures. The modified Gompertz model outperformed the others at 5, 15, and 20 °C, while Baranyi model was appropriate for 0 and 25 °C. The Huang model was optimal at 10 °C. No single model can give a consistently preferable goodness-of-fit for all growth data. The Gompertz model, with the smallest average values of RSS, AIC, MSE and the biggest pseudo-R² at all temperatures, is the most appropriate model to describe the growth of Pseudomonas of raw pork under pallet packaging.     

Validation of radio-frequency dielectric heating system for destruction of Cronobacter sakazakii and Salmonella species in nonfat dry milk

Cronobacter sakazakii and Salmonella species have been associated with human illnesses from consumption of contaminated nonfat dry milk (NDM), a key ingredient in powdered infant formula and many other foods. Cronobacter sakazakii and Salmonella spp. can survive the spray-drying process if milk is contaminated after pasteurization, and the dried product can be contaminated from environmental sources. Compared with conventional heating, radio-frequency dielectric heating (RFDH) is a faster and more uniform process for heating low-moisture foods. The objective of this study was to design an RFDH process to achieve target destruction (log reductions) of C. sakazakii and Salmonella spp. The thermal destruction (decimal reduction time; D-value) of C. sakazakii and Salmonella spp. in NDM (high-heat, HH; and low-heat, LH) was determined at 75, 80, 85, or 90°C using a thermal-death-time (TDT) disk method, and the z-values (the temperature increase required to obtain a decimal reduction of the D-value) were calculated. Time and temperature requirements to achieve specific destruction of the pathogens were calculated from the thermal destruction parameters, and the efficacy of the RFDH process was validated by heating NDM using RFDH to achieve the target temperatures and holding the product in a convection oven for the required period. Linear regression was used to determine the D-values and z-values. The D-values of C. sakazakii in HH- and LH-NDM were 24.86 and 23.0 min at 75°C, 13.75 and 7.52 min at 80°C, 8.0 and 6.03 min at 85°C, and 5.57 and 5.37 min at 90°C, respectively. The D-values of Salmonella spp. in HH- and LH-NDM were 23.02 and 24.94 min at 75°C, 10.45 and 12.54 min at 80°C, 8.63 and 8.68 min at 85°C, and 5.82 and 4.55 min at 90°C, respectively. The predicted and observed destruction of C. sakazakii and Salmonella spp. were in agreement, indicating that the behavior of the organisms was similar regardless of the heating system (conventional vs. RFDH). Radio-frequency dielectric heating can be used as a faster and more uniform heating method for NDM to achieve target temperatures for a postprocess lethality treatment of NDM before packaging.     

Thermal inactivation of Yersinia enterocolitica in pork slaughter plant scald tank water

The objective of this study was to establish the time–temperature combinations required to ensure the thermal inactivation of Yersinia enterocolitica during scalding of pork carcasses. A 2 strain cocktail of Y. enterocolitica (bioserotypes 2/O:5,27 and 1A/O:6,30) was heat treated at 50, 55 and 60 °C in samples of scald tank water obtained from a commercial pork slaughter plant. Samples were removed at regular intervals and surviving cells enumerated using (i) Cefsulodin–Irgasan–Novobiocin Agar (CIN) supplemented with ampicillin and arabinose and (ii) Tryptone Soya Agar (TSA), overlaid with CIN agar with ampicillin and arabinose. The data generated was used to estimate D- and z-values and the formula D_x = log^− 1(log D₆₀ − ((t₂ − t₁)/z)) was applied to calculate thermal death time–temperature combinations from 55 to 65 °C. D₅₀, D₅₅ and D₆₀-values of 45.9, 10.6 and 2.7 min were calculated from the cell counts obtained on CIN agar, respectively. The corresponding D-values calculated from the TSA/CIN counts were 45.1, 11 and 2.5 min, respectively. The z-value was 7.8. It was concluded that a time–temperature combination of 2.7 min at 60 °C is required to achieve a 1 log reduction in Y. enterocolitica in pork scald tank water. The predicted equivalent at 65 °C was 0.6 min. This study provides data and a model to enable pork processors to identify and apply parameters to limit the risk of carcass cross-contamination with Y. enterocolitica in pork carcass scald tanks.     

Growth of Clostridium perfringens from spore inocula in cooked cured beef: development of a predictive model

The objective of this study was to develop a model to predict the growth of C. perfringens from spores at temperatures applicable to the cooling of cooked cured meat products. C. perfringens growth from spores was not observed at a temperature of 12 °C for up to 3 weeks. The two parameters: germination, outgrowth, and lag (GOL) time and exponential growth rate, EGR, were determined using a function derived from mechanistic and stochastic considerations and the observed relative growths at specified times. A general model to predict the amount of relative growth for arbitrary temperature was determined by fitting the exponential growth rates to a square root Ratkowsky function, and assuming a constant ratio of GOL and generation times. The predicted relative growth is sensitive to the value of this ratio. A closed form equation was developed that can be used to estimate the relative growth for a general cooling scenario and determine a standard error of the estimate. The equation depends upon microbiological assumptions of the effect of history of the GOL times for gradual changes in temperature. Applying multivariate statistical procedures, a confidence interval was computed on the prediction of the amount of growth for a given temperature. The model predicts, for example, a relative growth of 3.17 with an upper 95% confidence limit of 8.50 when cooling the product from 51 to 11 °C in 8 h, assuming a log linear decline in temperature with time.     

Thermal inactivation of Salmonella spp. during conching

Although chocolate is a microbiologically stable product it has been described as a vehicle for Salmonella spp. Because of the low water activity (a_w) and the high fat content of chocolate Salmonella spp. shows an increased heat resistance, even during the thermal process of chocolate making. The aim of this study was to evaluate the thermal inactivation of Salmonella spp. during conching in various masses of chocolate and cocoa butter at different temperatures (50-90 °C). The effect of thermal treatment on Salmonella spp. was determined with the MPN (Most-Probable-Number) method. Results of thermal treatment showed approximate D-values for cocoa butter from D_50°C = 245 min to D_60°C = 306 min, for cocoa liquor from D_50°C = 999 min to D_90°C = 26 min and for dark chocolate of D_50°C = 1574 min. z-values were found to be z = 20 °C in cocoa liquor and z = 14 °C in dark chocolate. This study demonstrates that the conching process alone does not ensure the inactivation of Salmonella spp. in different chocolate masses and that an additional decontamination step at the beginning of the process as well as an HACCP concept is necessary during chocolate production to guarantee the absence of Salmonella spp. in chocolates and related products.     

A 3-D computational fluid dynamics model for forced air cooling of eggs placed in trays

Freshly laid shell eggs must be cooled quickly for controlling Salmonella Enteritidis (SE) growth. To fulfill a research need identified by Food Safety and Inspection Service (FSIS), a 3-D computational fluid dynamics (CFD) model was developed to predict the temperature of eggs placed on a tray (6 rows × 5 columns) under forced air cooling. The continuity, momentum, and energy equations were solved along with standard k − ε turbulence model using PHOENICS software. The model was validated by conducting experiments in a wind tunnel at various air temperatures (7-11 °C) and velocities (0.3-0.7 m/s). Root mean square error for predicting yolk temperatures was within 1 °C. Finally, the CFD model was integrated with a microbial growth model to estimate the risk of SE growth during cooling. This model can be incorporated into the FSIS risk assessment model for more accurate estimation of SE risk in shell eggs.     

Short communication: Antimicrobial effect of lactoferrin and its amidated and pepsin-digested derivatives against Salmonella Enteritidis and Pseudomonas fluorescens

The antimicrobial effect of bovine lactoferrin (LF) and its amidated and pepsin-digested derivatives, at concentrations varying from 0.25 to 20 mg/mL, against 3 Salmonella Enteritidis strains and 3 Pseudomonas fluorescens strains was investigated. Lactoferrin showed its maximum antimicrobial effect at 10 mg/mL against the 3 Salmonella strains, with reductions ranging from 1.3 to 2.0 log units, and the 3 Pseudomonas strains, with reductions ranging from 1.8 to 5.4 log units. In the case of amidated LF, the maximum effect on the 3 Salmonella strains was recorded at 0.25 mg/mL, with reductions in the range of 0.8 to 1.2 log units, whereas it was recorded at 1 mg/mL for the 3 Pseudomonas strains, with reductions in the range of 4.4 to 6.0 log units. Pepsin-digested LF showed its maximum antimicrobial effect at 1 mg/mL against the 3 Salmonella strains, with reductions ranging from 2.6 to 3.4 log units, and at 20 mg/mL against the 3 Pseudomonas strains, with reductions ranging from 4.5 to 5.4 log units. It is worth noting the pronounced effect (reductions exceeding 2.5 log units) of a low (1 mg/mL) concentration of pepsin-digested LF, which is naturally formed in the gastrointestinal tract, on Salmonella and Pseudomonas strains. A highly significant inverse correlation was found between capsule polysaccharide levels of bacterial strains and their lethality in the presence of different concentrations of amidated lactoferrin.     

Development of multiplex loop-mediated isothermal amplification-RFLP (mLAMP-RFLP) to detect Salmonella spp. and Shigella spp. in milk

A multiplex loop-mediated isothermal amplification-RFLP (mLAMP-RFLP) was developed and validated for simultaneous detection of Salmonella strains and Shigella strains in milk. In this system, two sets of LAMP primers were designed to specifically target invA of Salmonella spp. and ipaH of Shigella spp. Under isothermal conditions at 63 °C, ladder pattern of DNA bands could be amplified within 60 min in the presence of genomic DNAs of Salmonella strains and Shigella strains, which could be distinguished between Salmonella spp. and Shigella spp. simultaneously based on the different ladder pattern of DNA bands and subsequent restriction enzyme analysis. The overall analysis time was approximately 20 h including the enrichment of the bacterial cells, which greatly saved detection time. The sensitivity of mLAMP was found to be 100 fg DNA/tube with genomic DNAs of Salmonella strains and Shigella strains, comparatively, multiplex PCR was 1 pg DNA/tube. The mLAMP allowed the detection of milk sample artificially contaminated by Salmonella strains and Shigella strains at initial inoculation levels of approximate 5 CFU/10 mL. In conclusion, the mLAMP described here can potentially facilitate simultaneous monitoring of Salmonella and Shigella in a large number of food samples, which could be used as a primary screening method and as a supplement to classical detection method.     

Evaluation of the effects of ultraviolet light on bacterial contaminants inoculated into whole milk and colostrum,and on colostrum immunoglobulin G

Raw milk and colostrum can harbor dangerous microorganisms that can pose serious health risks for animals and humans. According to the USDA, more than 58% of calves in the United States are fed unpasteurized milk. The aim of this study was to evaluate the effect of UV light on reduction of bacteria in milk and colostrum, and on colostrum IgG. A pilot-scale UV light continuous (UVC) flow-through unit (45 J/cm²) was used to treat milk and colostrum. Colostrum and sterile whole milk were inoculated with Listeria innocua, Mycobacterium smegmatis, Salmonella serovar Typhimurium, Escherichia coli, Staphylococcus aureus, Streptococcus agalactiae, and Acinetobacter baumannii before being treated with UVC. During UVC treatment, samples were collected at 5 time points and bacteria were enumerated using selective media. The effect of UVC on IgG was evaluated using raw colostrum from a nearby dairy farm without the addition of bacteria. For each colostrum batch, samples were collected at several different time points and IgG was measured using ELISA. The UVC treatment of milk resulted in a significant final count (log cfu/mL) reduction of Listeria monocytogenes (3.2 ± 0.3 log cfu/mL reduction), Salmonella spp. (3.7 ± 0.2 log cfu/mL reduction), Escherichia coli (2.8 ± 0.2 log cfu/mL reduction), Staph. aureus (3.4 ± 0.3 log cfu/mL reduction), Streptococcus spp. (3.4 ± 0.4 log cfu/mL reduction), and A. baumannii (2.8 ± 0.2 log cfu/mL reduction). The UVC treatment of milk did not result in a significant final count (log cfu/mL) reduction for M. smegmatis (1.8 ± 0.5 log cfu/mL reduction). The UVC treatment of colostrum was significantly associated with a final reduction of bacterial count (log cfu/mL) of Listeria spp. (1.4 ± 0.3 log cfu/mL reduction), Salmonella spp. (1.0 ± 0.2 log cfu/mL reduction), and Acinetobacter spp. (1.1 ± 0.3 log cfu/mL reduction), but not of E. coli (0.5 ± 0.3 log cfu/mL reduction), Strep. agalactiae (0.8 ± 0.2 log cfu/mL reduction), and Staph. aureus (0.4 ± 0.2 log cfu/mL reduction). The UVC treatment of colostrum significantly decreased the IgG concentration, with an observed final mean IgG reduction of approximately 50%. Development of new methods to reduce bacterial contaminants in colostrum must take into consideration the barriers imposed by its opacity and organic components, and account for the incidental damage to IgG caused by manipulating colostrum.     

Salmonella surveillance and control at post-harvest in the Belgian pork meat chain

Salmonella remains the primary cause of reported bacterial food borne disease outbreaks in Belgium. Pork and pork products are recognized as one of the major sources of human salmonellosis. In contrast with the primary production and slaughterhouse phases of the pork meat production chain, only a few studies have focussed on the post-harvest stages. The goal of this study was to evaluate Salmonella and Escherichia coli contamination at the Belgian post-harvest stages. E. coli counts were estimated in order to evaluate the levels of faecal contamination. The results of bacteriological analysis from seven cutting plants, four meat-mincing plants and the four largest Belgian retailers were collected from official and self-monitoring controls. The prevalence of Salmonella in the cutting plants and meat-mincing plants ranged from 0% to 50%. The most frequently isolated serotype was Salmonella typhimurium. The prevalence in minced meat at retail level ranged from 0.3% to 4.3%. The levels of Salmonella contamination estimated from semi-quantitative analysis of data relating to carcasses, cuts of meat and minced meat were equal to −3.40 ± 2.04 log CFU/cm², −2.64 ± 1.76 log CFU/g and −2.35 ± 1.09 log CFU/g, respectively. The E. coli results in meat cuts and minced meat ranged from 0.21 ± 0.50 to 1.23 ± 0.89 log CFU/g and from 1.33 ± 0.58 to 2.78 ± 0.43 log CFU/g, respectively. The results showed that faecal contamination still needs to be reduced, especially in specific individual plants.     

Inactivation of Salmonella spp. on tomatoes by plant molecules

The efficacy of carvacrol (CAR), trans-cinnamaldehyde (TC), eugenol (EUG) and β-resorcylic acid (BR) as a wash treatment for reducing Salmonella spp. on tomatoes was investigated. Plum tomatoes inoculated with a six-serotype mixture of Salmonella (10⁸ CFU) were subjected to washing in sterile deionized water (control) or deionized water containing chlorine (100 ppm), CAR (0.25 and 0.75%), TC (0.5 and 0.75%), EUG (0.25 and 0.75%), or BR (0.75 and 1.0%) for 15 sec, 1 min, and 3 min. The plant molecules were more effective (P < 0.05) in reducing Salmonella on tomatoes compared to washing in water and chlorine. Both concentrations of CAR and TC, and 0.75% EUG decreased Salmonella counts on tomatoes by ~ 6.0 log CFU/ml at 1 min. Both concentrations of BR decreased the pathogen on tomatoes to undetectable levels at 3 min of exposure. Washing of tomatoes in deionized water and chlorine for 3 min reduced Salmonella by ca. 2.0 and 4.0 log CFU/ml, respectively. No Salmonella was detected in the wash water containing the plant molecules or chlorine, whereas a substantial population of the pathogen survived in the control wash water. Moreover, none of the dipping treatments had any effect on the red color of tomatoes (P > 0.05). Results indicate that CAR, TC, EUG and BR could effectively be used to kill Salmonella on tomatoes, but additional studies on sensory and quality characteristics of tomatoes treated with plant molecules are warranted.     

Behavior of Salmonella during fermentation,drying and storage of cocoa beans

Due to cocoa being considered a possible source of Salmonella contamination in chocolate, the behavior of Salmonella during some cocoa pre-processing stages (fermentation, drying and storage) was investigated. The fermentation process was carried out on a pilot scale (2 kg beans/box) for 7 days. Every day a fermentation box was inoculated with a Salmonella pool (ca. 4 log MPN/g). The results showed that Salmonella did not affect (P > 0.05) the growth of the main microorganism groups involved in cocoa fermentation. On the other hand, the pathogen was influenced (P < 0.05) by yeast, acetic acid bacteria and pH. In spite of Salmonella showing counts ≤ 1 log MPN/g in the first days, at the end of fermentation it grew in all samples, reaching counts as high as 7.49 log MPN/g. For drying and storage, cocoa beans were inoculated during the fermentation (experiment A) or during the drying (experiment B). In these stages the decline of the water activity affected the pathogen behavior. In experiment A during the drying, Salmonella count increased in most of the samples. In experiment B either a slight growth or no growth in the samples inoculated up to 48 h was observed, whereas the other samples showed reductions from the initial count. After 30 days of storage at room temperature, the water activity decreased to 0.68, and reductions of Salmonella ranged from 0.93 to 2.52 log MPN/g. Despite the reductions observed during the storage, the pathogen was detected even after 120 days. Therefore, the results showed that Salmonella growth or survival depends on when the contamination occurs.     

Effect of natural microbiota on growth of Salmonella spp. in fresh pork – A predictive microbiology approach

This study was undertaken to model and predict growth of Salmonella and the dominating natural microbiota, and their interaction in ground pork. Growth of Salmonella in sterile ground pork at constant temperatures between 4 °C and 38 °C was quantified and used for developing predictive models for lag time, max. specific growth rate and max. population density. Data from literature were used to develop growth models for the natural pork microbiota. Challenge tests at temperatures from 9.4 to 24.1 °C and with Salmonella inoculated in ground pork were used for evaluation of interaction models. The existing Jameson-effect and Lotka–Volterra species interaction models and a new expanded Jameson-effect model were evaluated. F-test indicated lack-of-fit for the classical Jameson-effect model at all of the tested temperatures and at 14.1–20.2 °C this was caused by continued growth of Salmonella after the natural microbiota had reached their max. population density. The new expanded Jameson-effect model and the Lotka–Volterra model performed better and appropriately described the continued but reduced growth of Salmonella after the natural microbiota had reached their max. population density. The expanded Jameson-effect model is a new and simple species interaction model, which performed as well as the more complex Lotka–Volterra model.     

Prediction of pathogen growth on iceberg lettuce under real temperature history during distribution from farm to table

The growth of pathogenic bacteria Escherichia coli O157:H7, Salmonella spp., and Listeria monocytogenes on iceberg lettuce under constant and fluctuating temperatures was modelled in order to estimate the microbial safety of this vegetable during distribution from the farm to the table. Firstly, we examined pathogen growth on lettuce at constant temperatures, ranging from 5 to 25 degrees C, and then we obtained the growth kinetic parameters (lag time, maximum growth rate (micro(max)), and maximum population density (MPD)) using the Baranyi primary growth model. The parameters were similar to those predicted by the pathogen modelling program (PMP), with the exception of MPD. The MPD of each pathogen on lettuce was 2-4 log(10) CFU/g lower than that predicted by PMP. Furthermore, the MPD of pathogens decreased with decreasing temperature. The relationship between mu(max) and temperature was linear in accordance with Ratkowsky secondary model as was the relationship between the MPD and temperature. Predictions of pathogen growth under fluctuating temperature used the Baranyi primary microbial growth model along with the Ratkowsky secondary model and MPD equation. The fluctuating temperature profile used in this study was the real temperature history measured during distribution from the field at harvesting to the retail store. Overall predictions for each pathogen agreed well with observed viable counts in most cases. The bias and root mean square error (RMSE) of the prediction were small. The prediction in which mu(max) was based on PMP showed a trend of overestimation relative to prediction based on lettuce. However, the prediction concerning E. coli O157:H7 and Salmonella spp. on lettuce greatly overestimated growth in the case of a temperature history starting relatively high, such as 25 degrees C for 5 h. In contrast, the overall prediction of L. monocytogenes under the same circumstances agreed with the observed data.     

Escherichia coli population heterogeneity: subpopulation dynamics at super-optimal temperatures

In the past years, we explored the dynamics of Escherichia coli K12 at super-optimal temperatures under static and dynamic temperature conditions ( [Van Derlinden et?al., 2008b], [Van Derlinden et?al., 2009] and [Van Derlinden et?al., 2010]). Disturbed sigmoid growth curves, i.e., a sequence of growth, inactivation and re-growth, were observed, especially close to the maximum growth temperature. Based on the limited set of experiments (i.e., 2 static temperatures and 2 dynamic temperature profiles), the irregular growth curves were explained by postulating the co-existence of two subpopulations: a more resistant, growing population and a temperature sensitive, inactivating population.In this study, the dynamics of the two subpopulations are studied rigorously at 11 constant temperature levels in the region between 45 °C and 46 °C, with at least five repetitions per temperature.At all temperatures, the total population follows a sequence of growth, inactivation and re-growth. The sequence of different stages in the growth curves can be explained by the two subpopulations. The first growth phase and the inactivation phase reflect the presence of the sensitive subpopulation. Hereafter, the population’s dynamics are dominated by the growth of the resistant subpopulation. Generally, cell counts are characterized by a large variability.The dynamics of the two subpopulations are carefully analyzed using a heterogeneous subpopulation type model to study the relation between the kinetic parameters of the two subpopulations and temperature, and to evaluate if the fraction d of resistant cells varies with temperature. Results indicate that the growth rate of the sensitive subpopulation decreases with increasing temperature within the range of 45-46 °C. Furthermore, results point in the direction that the duration of this initial growth phase is approximately constant, i.e., around 2 h. Possibly, the stress resistance of the cells decreases after a certain period because the metabolism is fully adapted to exponential growth. Also, the growth rate of the resistant subpopulation decreases with increasing temperature. Due to the extreme variability in the cell density data, derivation of accurate relations was not possible. From the heterogeneous model implementations, given the experimental set-up, both a constant d value and a temperature dependent d value seem plausible.     

In vitro inactivation of Escherichia coli, Staphylococcus aureus and Salmonella spp. using slightly acidic electrolyzed water

In the current study, the effectiveness of slightly acidic electrolyzed water (SAEW) on an in vitro inactivation of Escherichia coli (E. coli), Staphylococcus aureus (S. aureus) and Salmonella spp. was evaluated and compared with other sanitizers. SAEW (pH 5.6, 23 mg/l available chlorine concentration; ACC; and 940 mV oxidation reduction potential; ORP) was generated by electrolysis of dilute solution of HCl (2%) in a chamber of a non-membrane electrolytic cell. One milliliter of bacteria suspension (ca. 10–11 log₁₀CFU/ml) was mixed with 9 ml of SAEW, strong acidic electrolyzed water (StAEW; ca. 50 mg/l ACC), sodium hypochlorite solution (NaOCl; ca.120 mg/l ACC) and distilled water (DW) as control and treated for 60 s. SAEW effectively reduced the population of E. coli, S. aureus and Salmonella spp. by 5.1, 4.8, and 5.2 log₁₀CFU/ml. Although, ACC of SAEW was more than 5 times lower than that of NaOCl solution, they showed no significant bactericidal difference (p > 0.05). However, the bactericidal effect of StAEW was significantly higher (p < 0.05) than SAEW and NaOCl solution in all cases. When tested with each individual test solution, E. coli, S. aureus and Salmonella spp. reductions were not significantly different (p > 0.05). These findings indicate that SAEW with low available chlorine concentration can equally inactivate E. coli, S. aureus and Salmonella spp. as NaOCl solution and therefore SAEW shows a high potential of application in agriculture and food industry as an environmentally friendly disinfection agent.