Effect of inoculum size, bacterial species, type of surfaces and contact time to the transfer of foodborne pathogens from inoculated to non-inoculated beef fillets via food processing surfaces.

The objective of the present study was to determine the factors affecting the transfer of foodborne pathogens from inoculated beef fillets to non-inoculated ones, through food processing surfaces. Three different levels of inoculation of beef fillets surface were prepared: a high one of approximately 107 CFU/cm2, a medium one of 105 CFU/cm2 and a low one of 103 CFU/cm2, using mixed-strains of Listeria monocytogenes, or Salmonella enterica Typhimurium, or Escherichia coli O157:H7. The inoculated fillets were then placed on 3 different types of surfaces (stainless steel-SS, polyethylene-PE and wood-WD), for 1 or 15 min. Subsequently, these fillets were removed from the cutting boards and six sequential non-inoculated fillets were placed on the same surfaces for the same period of time. All non-inoculated fillets were contaminated with a progressive reduction trend of each pathogen's population level from the inoculated fillets to the sixth non-inoculated ones that got in contact with the surfaces, and regardless the initial inoculum, a reduction of approximately 2 log CFU/g between inoculated and 1st non-inoculated fillet was observed. S. Typhimurium was transferred at lower mean population (2.39 log CFU/g) to contaminated fillets than E. coli O157:H7 (2.93 log CFU/g), followed by L. monocytogenes (3.12 log CFU/g; P < 0.05). Wooden surfaces (2.77 log CFU/g) enhanced the transfer of bacteria to subsequent fillets compared to other materials (2.66 log CFU/g for SS and PE; P < 0.05). Cross-contamination between meat and surfaces is a multifactorial process strongly depended on the species, initial contamination level, kind of surface, contact time and the number of subsequent fillet, according to analysis of variance. Thus, quantifying the cross-contamination risk associated with various steps of meat processing and food establishments or households can provide a scientific basis for risk management of such products.

Effect of the substrate's microstructure on the growth of Listeria monocytogenes.

The effect of the microstructure of the medium on the growth of the food-borne pathogen Listeria monocytogenes was studied. The pathogen's growth kinetics was evaluated using liquid substrates and gels formed from different concentrations of sodium alginate (3.0% w/w) and gelatin (0-30.0% w/w). These results were further verified using a model dairy product with solid concentrations varying from 10.0 to 40.0% w/w. The pathogen's growth was faster in the liquid media than in the gels regardless of the gelling agent employed. The substrate's microstructure, apart from altering the growth pattern from planktonic to colonial, resulted in microbial growth suppression; however, each system affected the microorganism's growth in a different way. The suppressing effect of the substrate's microstructure on microbial growth was also dependent on temperature, while the presence of glucose in the solid medium accelerated microbial growth, thus reducing substantially the difference in growth kinetics between the gels and the liquid media. Any increase in the hydrocolloid concentration, which was also reflected in the rheological properties of the structured samples, resulted in a reduction of growth rate and in an increase of the lag phase of the pathogen. Overall, the gelation of the medium was found to exert a stress on the microorganism since the sol-gel transition, when the pathogen was already at the exponential growth phase, resulted in an additional lag phase or a decrease in the growth rate. The relationship between maximum specific growth rate and loss tangent of the gels (tanδ=G″/G') was explored, pointing to the possible use of a single structural parameter to describe food matrix effects on microbial growth kinetics.

Use of marination for controlling Salmonella enterica and Listeria monocytogenes in raw beef.

The effect of marination on the survival and growth of the pathogens Salmonella enterica and Listeria monocytogenes on beef pieces was investigated. Five marinades were used: soy sauce base marinade without (SB) or with lactic acid (SBLA), red wine base marinade without (WB) or with 0.5% v/v oregano essential oil (WBO), and sterile saline used as control (C). Inoculated fresh beef pieces were marinated for 18 h at 5 °C, removed from the marinade and subjected to storage trials at 5 °C and 15 °C. Heat inactivation studies were also performed on the isolates after exposure to the marinades to determine if marination affects heat resistance of the pathogens. The marinades with antimicrobials caused a significant decrease in viable count of the pathogens during marinations at 5 °C for 18 h of up to 2.1 and 3.4 log cfu cm(-2) for Salmonella and L. monocytogenes, respectively. Marinades without antimicrobials were less bactericidal resulting to reductions ranging from 0.3 to 0.4 and 1.3 to 2.0 log cfu cm(-2) for Salmonella and L. monocytogenes, respectively. Growth of L. monocytogenes was observed in the controls at both tested temperatures, while growth of Salmonella was observed in the controls stored at 15 °C. No growth of the pathogens was observed in any of the marinated samples at both temperature tested. No significant changes of heat resistance of the tested pathogens after exposure to the marinades were observed demonstrating the enhanced safety of the marinated beef product.

Transfer of foodborne pathogenic bacteria to non-inoculated beef fillets through meat mincing machine.

The aim of this study was to evaluate the transfer of pathogens population to non-inoculated beef fillets through meat mincing machine. In this regard, cocktails of mixed strain cultures of each Listeria monocytogenes, Salmonella enterica ser. Typhimurium and Escherichia coli O157:H7 were used for the inoculation of beef fillets. Three different initial inoculum sizes (3, 5, or 7 log CFU/g) were tested. The inoculated beef fillets passed through meat mincing machine and then, six non-inoculated beef fillets passed in sequence through the same mincing machine without sanitation. The population of each pathogen was measured. It was evident that, all non-inoculated beef fillets were contaminated through mincing with all pathogens, regardless the inoculum levels used. This observation can be used to cover knowledge gaps in risk assessments since indicates the potential of pathogen contamination and provides significant insights for the risk estimation related to cross-contamination, aiming thus to food safety enhancement.

Efficacies of soy sauce and wine base marinades for controlling spoilage of raw beef.

Fresh beef slices were marinated by immersion in marinades based on soy sauce without (SB) or with lactic acid (SBLA) or red wine base without (WB) or with 0.5% v/v oregano essential oil (WBO). For control samples (immersed in saline), a mean increase of 0.9log CFU/cm(2) in total viable counts (TVCs) occurred during the 24h treatment. During marination with WB and SB, mean TVC decreased by 0.7 and 0.3log CFU/cm(2), respectively. The mean decrease in TVC for samples marinated in WBO or SBLA was 1.2log CFU/cm(2). Subsequent storage of beef resulted in a rapid increase of TVC in control samples, to ≥9.5log CFU/cm(2) after 8 days at 5°C or 3 days at 15°C. Significant (P<0.05) microbial growth occurred in marinated samples stored at 5°C. During storage at 15°C TVC increased in only WB samples but the final numbers of 5.9log CFU/cm(2) were significantly lower (P<0.05) than the numbers in the control. Results similar to those for TVC were observed for Pseudomonas spp. All marinades also gave meat with significant lower TBARS values than the controls. There were no significant differences (P>0.05) in the toughness of the marinated samples compared to the control, except for SBLA samples which had significantly higher (P<0.05) shear force values. Marination with soy sauce or red wine marinades can evidently control microbial spoilage and oxidation of meat.

Effect of microbial cell-free meat extract on the growth of spoilage bacteria.

AIMS: This study examined the effect of microbial cell-free meat extract (CFME) derived from spoiled meat, in which quorum sensing (QS) compounds were present, on the growth kinetics (lag phase, and growth rate) of two spoilage bacteria, Pseudomonas fluorescens and Serratia marcescens. METHODS AND RESULTS: Aliquots of CFME from spoiled meat were transferred to Brain Heart Infusion broth inoculated with 10(3) CFU ml(-1) of 18 h cultures of Ps. fluorescens or Ser. marcescens, both fresh meat isolates; CFME derived from unspoiled fresh meat ('clean' meat) served as a control. Changes in impedance measurements were monitored for 48 h, and the detection time (Tdet) was recorded. It was found that in the absence of CFME containing QS compounds the Tdet was shorter (P < 0.05) than that in broth samples with added CFME from spoiled meat. The rate of growth of Ps. fluorescens, recorded as the maximum slope rate of conductance changes (MSrCC), after Tdet, was higher (P < 0.05) in samples with CFME containing QS compounds compared to samples without CFME or CFME derived from 'clean' meat. Similar results in MSrCC of impedance changes were obtained for Ser. marcescens. CONCLUSIONS: The study indicated that the growth rate (expressed in MSrCC units) of meat spoilage bacteria in vitro was enhanced in samples supplemented with CFME containing QS compounds compared to control samples (i.e., without CFME or with CFME from 'clean' meat). This behaviour may explain the dominant role of these two bacteria in the spoilage of meat. SIGNIFICANCE AND IMPACT OF THE STUDY: These results illustrate the potential effect of signalling compounds released during storage of meat on the behaviour of meat spoilage bacteria. Understanding such interactions may assist in the control of fresh meat quality and the extension of its shelf life.

Prevalence and concentration of verocytotoxigenic Escherichia coli, Salmonella enterica and Listeria monocytogenes in the beef production chain: a review.

This review examines the prevalence of three important pathogens, verocytotoxigenic Escherichia coli (VTEC), Salmonella enterica and Listeria monocytogenes, in cattle and beef from the farm to the final, ready-to-eat product. Factors affecting prevalence of pathogens in the beef chain, such as the season and cattle rearing method, are examined. Data from many key surveys are summarized in table form. The observed prevalence of pathogens in cattle and beef varies considerably from survey to survey. An indication of relative prevalence of pathogens at different stages can be obtained by calculating average prevalences observed over multiple surveys, weighted by sample number. Based on the data presented in the tables in this review, for E. coli O157 at selected processing stages the mean prevalences (and range of means from individual surveys) are faeces 6.2% (0.0-57%), hides 44% (7.3-76%), chilled carcasses 0.3% (0.0-0.5%), and raw beef products 1.2% (0.0-17%). For Salmonella the mean prevalence data are faeces 2.9% (0.0-5.5%), hides 60% (15-71%), chilled carcasses 1.3% (0.2-6.0%), and raw beef products 3.8% (0.0-7.5%). For L. monocytogenes the mean prevalence data are faeces 19% (4.8-29%), hides 12% (10-13%), and raw beef products 10% (1.6-24%). Seasonal variation was evident in many surveys, faecal prevalences of E. coli O157 and Salmonella generally being higher in the warmer months. The influence of animal type, animal age, feed and housing on pathogen carriage has also been examined. The significance of non-O157 serotypes of VTEC and their detection and classification are discussed.

A study on the variability in the growth limits of individual cells and its effect on the behavior of microbial populations.

The NaCl growth limits of individual Salmonella enteritidis cells were studied. Six different levels of the pathogen (from 10(1) to 10(6) cells) were inoculated on tryptic soy agar plates at NaCl concentrations ranging from 0.5 to 8% and pH values of 5.0, 5.5 or 7.3. The NaCl growth limits of individual cells were estimated from the ratio between the number of colonies formed at each NaCl concentration interval and the number of cells initially inoculated, based on the assumption that each colony was derived from a single cell. The results showed that, as the NaCl concentration increased and the pH decreases, the number of cells able to grow and form a colony gradually decreased indicating variability in the growth limits of individual cells. The distribution of single-cell growth limits was used to predict the behavior of microbial populations. The results showed that as the inoculum size increased the NaCl growth limits of S. enteritidis increased and became less variable. Furthermore, at conditions close to the boundary of growth the presence of a non-growing fraction in the population resulted in a delay of population growth, which we called "pseudo-lag". The extent of "pseudo-lag" is determined by the environment while its variability is affected by both the inoculum size and the growth conditions. At growth-limiting conditions the total "apparent lag" of the population is a convolution of the "pseudo-lag" and the "physiological lag" of the growing cells. The data on the variability in the growth limits of single microbial cells presented in this work stress the need for stochastic approaches in quantitative microbiology especially at environments close to the boundary of growth.

Control of spoilage microorganisms in minced pork by a self-developed modified atmosphere induced by the respiratory activity of meat microflora.

The changes in microbial flora of minced pork during aerobic storage at 0, 5, 10 and 15 degrees C were studied. Minced pork samples (100g) were packed using two types of packaging films: (a) a common food film with high permeability (HPF) and (b) a film with low permeability (LPF). The respiratory activity of meat microflora and the use of a LPF resulted in a modified atmosphere in the package headspace developed during storage. Oxygen concentration decreased from 18.7% (after packaging) to 7% (after 15 days of storage) in packages with LPF, stored at 0 degrees C, while CO(2) increased from 3% to 10.5%, respectively. On the contrary, no significant atmosphere changes were observed during storage of HPF packages. The self-developed modified atmosphere in LPF packages resulted in a significant inhibition of pseudomonad growth which was more pronounced at low storage temperatures. For example, during storage at 0 degrees C, the growth rate of pseudomonads in meat packed with LPF was reduced by 48.7% compared to HPF. At 10 degrees C the latter reduction decreased to 13.7%. LPF packaging was also found to inhibit the growth of Brochothrix thermosphacta but this inhibition was weaker compared to pseudomonads. The effect of storage temperature on the growth rate of pseudomonads and B. thermosphacta in minced pork packed with the different films was modeled using an Arrhenius equation. For both bacteria, the activation energy was higher for LPF packaging. This can be attributed to the increased inhibitory effect of the modified atmosphere at lower storage temperature. The Arrhenius model was further used to evaluate the effect of temperature on the time required by the two bacteria to reach a spoilage level of 10(7)CFU/g. The results showed that when LPF packaging is combined with effective temperature control the time-to-spoilage can be significantly extended compared to HPF packaging.

A study on the kinetic behavior of Listeria monocytogenes in ice cream stored under static and dynamic chilling and freezing conditions.

The kinetic behavior of Listeria monocytogenes in 2 commercial ice cream products (A and B) that were inoculated and stored under static chilling (4 to 16 degrees C), static freezing (-5 to -33 degrees C), dynamic chilling, and dynamic chilling-freezing conditions was studied, simulating conditions of the aging process and of normal or abuse conditions during distribution and storage. The ice cream products A and B had different compositions but similar pH (6.50 and 6.67, respectively) and water activity (0.957 and 0.965, respectively) values. For both chilling and freezing conditions, the kinetic behavior of the pathogen was similar in the 2 products, indicating that the pH and water activity, together with temperature, were the main factors controlling growth. Under chilling conditions, L. monocytogenes grew well at all temperatures tested. Under freezing conditions, no significant changes in the population of the pathogen were observed throughout a 90-d storage period for either of the inoculum levels tested (10(3) and 10(6) cfu/g). Growth data from chilled storage conditions were fitted to a mathematical model, and the calculated maximum specific growth rate was modeled as a function of temperature by using a square root model. The model was further validated under dynamic chilling and dynamic chilling-freezing conditions by using 4 different storage temperature scenarios. Under dynamic chilling conditions, the model accurately predicted the growth of the pathogen in both products, with 99.5% of the predictions lying within the +/- 20% relative error zone. The results from the chilling-freezing storage experiments showed that the pathogen was able to initiate growth within a very short time after a temperature upshift from freezing to chilling temperatures. This indicates that the freezing conditions did not cause a severe stress in L. monocytogenes cells capable of leading to a significant "additional" lag phase during the subsequent growth of the pathogen at chilling conditions. As a result, the application of the model at chilling-freezing conditions resulted in satisfactory performance, with 98.3% of the predictions lying within the +/- 20% relative error zone. The present study provides useful data for understanding the behavior of L. monocytogenes in ice cream stored under single or combined chilling and freezing conditions. In addition, the study showed that such data can be expressed in quantitative terms via the application of mathematical models, which can be used by the dairy industry as effective tools for predicting the behavior of the pathogen during the manufacture, distribution, and storage of ice cream products.

Dynamic modeling of Listeria monocytogenes growth in pasteurized milk.

AIMS: The development and validation of a dynamic model for predicting Listeria monocytogenes growth in pasteurized milk stored at both static and dynamic temperature conditions. METHODS AND RESULTS: Growth of inoculated L. monocytogenes in a commercial pasteurized whole milk product was monitored at various isothermal conditions from 1.5 to 16 degrees C. The kinetic parameters of the pathogen were modelled as a function of temperature using a square root type model, which was further validated using data from 92 published growth curves from eight different milk products. Compared to four published models for L. monocytogenes growth, the model developed in this study performed better, with a per cent discrepancy and bias of 49.1 and -1.01%, respectively. The performance of the model in predicting growth at dynamic temperature conditions was evaluated at four different fluctuating temperature scenarios with periodic temperature changes from -2 to 16 degrees C. The prediction of growth at dynamic storage temperature was based on the square root model in conjunction with the differential equations of the Baranyi and Roberts model, which were numerically integrated with respect to time. The per cent relative errors between the observed and the predicted growth of L. monocytogenes were less than 10% for all temperature scenarios tested. CONCLUSIONS: Available models from experiments conducted in laboratory media may result in significant overestimation of L. monocytogenes growth in pasteurized milk because they do not take into account factors such as milk composition (e.g. natural antimicrobial compounds present in milk) and the interactions of the pathogen with the natural microflora. The product-targeted model developed in the present study showed a high performance in predicting growth of L. monocytogenes in pasteurized milk under both static and dynamic temperature conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: Temperature fluctuations often occur during the transportation and storage of pasteurized milk. A high performance, dynamic model for the growth of L. monocytogenes can be a useful tool for effective management and optimization of product safety and can lead to more realistic estimations of pasteurized-milk related safety risks.

Development of a microbial model for the combined effect of temperature and pH on spoilage of ground meat, and validation of the model under dynamic temperature conditions.

The changes in microbial flora and sensory characteristics of fresh ground meat (beef and pork) with pH values ranging from 5.34 to 6.13 were monitored at different isothermal storage temperatures (0 to 20 degrees C) under aerobic conditions. At all conditions tested, pseudomonads were the predominant bacteria, followed by Brochothrix thermosphacta, while the other members of the microbial association (e.g., lactic acid bacteria and Enterobacteriaceae) remained at lower levels. The results from microbiological and sensory analysis showed that changes in pseudomonad populations followed closely sensory changes during storage and could be used as a good index for spoilage of aerobically stored ground meat. The kinetic parameters (maximum specific growth rate [mu(max)] and the duration of lag phase [lambda]) of the spoilage bacteria were modeled by using a modified Arrhenius equation for the combined effect of temperature and pH. Meat pH affected growth of all spoilage bacteria except that of lactic acid bacteria. The "adaptation work," characterized by the product of mu(max) and lambda(mu(max) x lambda) was found to be unaffected by temperature for all tested bacteria but was affected by pH for pseudomonads and B. thermosphacta. For the latter bacteria, a negative linear correlation between ln(mu(max) x lambda) and meat pH was observed. The developed models were further validated under dynamic temperature conditions using different fluctuating temperatures. Graphical comparison between predicted and observed growth and the examination of the relative errors of predictions showed that the model predicted satisfactorily growth under dynamic conditions. Predicted shelf life based on pseudomonads growth was slightly shorter than shelf life observed by sensory analysis with a mean difference of 13.1%. The present study provides a "ready-to-use," well-validated model for predicting spoilage of aerobically stored ground meat. The use of the model by the meat industry can lead to effective management systems for the optimization of meat quality.

Field evaluation of the application of time temperature integrators for monitoring fish quality in the chill chain.

The applicability of time temperature integrators (TTI) as effective tools of chill chain monitoring was assessed. Validated kinetic models of pseudomonads growth of Mediterranean, marine-cultured chilled gilt-head seabream (Sparus aurata) and full knowledge of the response of suitable enzymatic TTI are the basis of the TTI application algorithm. This scheme was evaluated through a controlled field test of exported fish, from harvest to final consumption. Response of TTI attached on different locations of packages was compared to actual temperature recording. Data that could not be obtained during the actual field test, such as microbiological or sensory tests of fish at intermediate points of the chain, were measured in a replicate laboratory study, simulating the handling of products and the real time-temperature profiles of the field test. The conducted field tests showed the applicability and usefulness of TTI monitoring of the fish chill chain, elucidating also the practical difficulties and limitations, that need to be addressed for expanding TTI use as a reliable management tool.

Development of a Safety Monitoring and Assurance System for chilled food products.

The principles of a novel chill chain management policy, coded Safety Monitoring and Assurance System (SMAS) for the optimisation of the distribution of chilled food products within the chill chain are developed. In this system, a new approach based on actual risk evaluation at important points of the chill chain is used in order to promote products to the next stage of distribution. This evaluation based on product's time-temperature history, variation in product's characteristics (e.g. a(w), pH, etc.), and the use of predictive models for the growth of food pathogens, allows to give priority to products in such a way that risk at consumption time is minimized. The effectiveness of SMAS was evaluated against the First In First Out (FIFO) approach, the current method for food distribution, in a case study on the risk of listeriosis of cooked ham using the Monte Carlo simulation technique. Furthermore, the two approaches were compared for their effect on the quality of the products in terms of remaining shelf life at the time of consumption. The results showed that following the SMAS approach the risk of listerisosis is significantly lower while the spoiled products at the time of consumption are significantly reduced compared to FIFO approach.

Comparative acid stress response of Listeria monocytogenes, Escherichia coli O157:H7 and Salmonella Typhimurium after habituation at different pH conditions.

AIMS: The aim of the study was to evaluate the effect of habituation at different pH conditions on the acid resistance of Listeria monocytogenes, Escherichia coli O157:H7 and Salmonella enterica serotype Typhimurium, and to identify potential differences between the adaptive responses of the three pathogens. METHODS: Stationary phase cells of L. monocytogenes, E. coli O157:H7 and S. Typhimurium, grown in glucose-free media, were exposed to pH 3.5 broth directly or after habituation for 90 min at various pH conditions from 4.0 to 6.0. Survivors at pH 3.5 were determined by plating on tryptic soy agar and incubating at 30 degrees C for 48 h. The kinetics (death rate) of the pathogens at pH 3.5 was calculated by fitting the data to an exponential model. RESULTS: Habituation to acidic environments provided protection of the pathogens against lethal acid conditions. This acid protection, however, was found to be pH dependent. For example, for E. coli O157:H7 an increased acid resistance was observed after habituation at a pH range from 4.0 to 5.5, while the maximum acid tolerance was induced at pH 5.0. Furthermore, the effect of low pH habituation was different among pathogens. For L. monocytogenes, E. coli O157:H7 and S. Typhimurium, the pH range within which habituation resulted to increased acid resistance was 5.0-6.0, 4.0-5.5 and 4.0-5.0, respectively, while the maximum acid tolerance was induced after habituation at pH 5.5, 5.0 and 4.5, respectively. SIGNIFICANCE: Acid stress conditions are common within current food processing technologies. The information on adaptive responses of L. monocytogenes, E. coli O157:H7 and S. Typhimurium after habituation to different pH environments provided in the present study, could lead to a more realistic evaluation of food safety concerns and to a better selection of processes in order to avoid adaptation phenomena and to minimize the potential for food safety risks.

Application of shelf life decision system (SLDS) to marine cultured fish quality.

Growth of natural microflora of marine cultured, air-packed, sea bass (Dichentrachus labrax) was studied at isothermal conditions in the 0-15 degrees C range and kinetically modelled using the four-parameter Logistic equation. Sensory shelf life was correlated to pseudomonad population and sensory acceptability was correlated to a pseudomonad level, Ns, of 10(7). The variability of their initial population was quantitatively shown and a conductance-based rapid method specific to sea bass pseudomonad enumeration was established as a practical means of N0 determination, required in shelf life predictions. Kinetic models, shelf life correlations and N0 data were incorporated into the shelf life decision system (SLDS) shown to be an effective tool for marine cultured sea bass chill chain management leading to optimization of quality of the fish at consumer's end.

Development and assessment of an intelligent shelf life decision system for quality optimization of the food chill chain.

The principles of application of a Shelf Life Decision System (SLDS) for the optimization of the distribution of chilled fresh and minimally processed food products are developed. The SLDS integrates predictive kinetic models of food spoilage, data on initial quality from rapid techniques, and the capacity to continuously monitor temperature history of the food product with Time Temperature Integrators (TTIs) into an effective chill chain management tool that leads to an improved narrow distribution of quality at consumption time, effectively reducing the probability of products consumed past shelf life end. The applicability and effectiveness of the SLDS is demonstrated and evaluated based on actual food spoilage and TTI kinetics and chill chain data employing the Monte Carlo simulation method.

Predictive modeling of the shelf life of fish under nonisothermal conditions.

The behavior of the natural microflora of Mediterannean gilt-head seabream (Sparus aurata) was monitored during aerobic storage at different isothermal conditions from 0 to 15 degrees C. The growth data of pseudomonads, established as the specific spoilage organisms of aerobically stored gilt-head seabream, combined with data from previously published experiments, were used to model the effect of temperature on pseudomonad growth using a Belehradek type model. The nominal minimum temperature parameters of the Belehradek model (T(min)) for the maximum specific growth rate (micro(max)) and the lag phase (t(Lag)) were determined to be -11.8 and -12.8 degrees C, respectively. The applicability of the model in predicting pseudomonad growth on fish at fluctuating temperatures was evaluated by comparing predictions with observed growth in experiments under dynamic conditions. Temperature scenarios designed in the laboratory and simulation of real temperature profiles observed in the fish chill chain were used. Bias and accuracy factors were used as comparison indices and ranged from 0.91 to 1.17 and from 1.11 to 1.17, respectively. The average percent difference between shelf life predicted based on pseudomonad growth and shelf life experimentally determined by sensory analysis for all temperature profiles tested was 5.8%, indicating that the model is able to predict accurately fish quality in real-world conditions.

Comparison of maximum specific growth rates and lag times estimated from absorbance and viable count data by different mathematical models.

Maximum specific growth rate (mu(max)) and lag time (lambda) were estimated from viable count and absorbance data and compared for different microorganisms, incubation systems and growth conditions. Data from 176 growth curves and 120 absorbance detection times of serially diluted cultures were evaluated using different mathematical growth models. Accurate estimates of mu(max) and lambda were obtained from individual absorbance growth curves by using the Richard model, with values of the parameter m fixed to 0.5, 1.0 or 2.0 to describing different degrees of growth dampening, as well as from absorbance detection times of serially diluted cultures. It is suggested to apply the two techniques complementarily for accurate, rapid and inexpensive estimation of microbial growth parameter values from absorbance data. In contrast, considerable limitations were demonstrated for the ability of the Exponential, the Gompertz and the Logistic models to estimate mu(max) and lambda values accurately from absorbance data. Limitations of these models were revealed due the wide range of growth conditions studies.

Application of a systematic experimental procedure to develop a microbial model for rapid fish shelf life predictions.

A systematic experimental procedure for fish shelf-life modelling was used to develop a model for predicting the quality of fish in the chill chain. For this, the growth of the naturally occurring bacteria pseudomonads, Shewanella putrefaciens, Enterobacteriaceae, lactic acid bacteria and yeasts, on gilt-head seabream (Sparus aurata), was studied at temperatures from 0 to 15 degrees C. The results from the microbiological, organoleptical and chemical analysis conducted on naturally contaminated fish as well as on inoculated sterile fish blocks identified pseudomonads as a good spoilage index. Growth of pseudomonads was modelled as a function of storage temperature and correlated to organoleptical shelf life. To reduce the time required for the enumeration of the initial pseudomonads number, which is crucial information for shelf life prediction, a conductance assay was established. Compared with the conventional microbiological tests, this method gave results in one-fourth of the time.