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There is a lack of information about Salmonella enterica strains under acidic conditions and their association with their genome. This study characterized intraspecies variability in the growth of 167 S. enterica isolates under two acid conditions (pH 4 and 5) and linked to the whole genome sequencing (WGS) data. A total of 1002 curves for each condition were obtained using turbidimetry measurements, and Baranyi and Roberts model was used to estimate the maximum rate of change (rcmax; OD600 nm h−1). Strains were categorized into slow, intermediate, and fast; and associations with their WGS data were performed. Huge variability in rcmax¯$\overline {{\mathrm{r}}{{{\mathrm{c}}}_{{\mathrm{max}}}}} $ was observed at both conditions (pH 5 = 0.016–0.066 OD600nm h−1 and pH 4 = 0.003–0.028 OD600nm h−1). The majority of isolates was classified as intermediate rcmax¯$\overline {{\mathrm{r}}{{{\mathrm{c}}}_{{\mathrm{max}}}}} $ (59.5% at pH 5 and 46.1% at pH 4). Strains classified as fast had a low frequency of allABCD genes at both pHs, and any of them having the presence of pefABCD, spvBCR, aadA2, dfrA12, and gyrA_D87G genes were linked to virulence or antimicrobial resistance. This study suggests that strains with fast capacity for growth under acidic conditions could have a fitness cost in their virulence or resistance potential. Practical Application: Data presented in this study could be used to select representative strains to evaluate the exposure assessment in different food items, mainly the growth and survival in acidic foods. [ABSTRACT FROM AUTHOR]

To estimate human exposure to Salmonella enterica, it is essential to understand the pathogen distribution and characteristics. Prevalence and concentration of S. enterica were determined in mango, tomato, and raw chicken samples purchased in three states (Aguascalientes, Querétaro, and Guadalajara) located in the central region of Mexico during two seasons. In addition, S. enterica isolates were characterized by absence/presence of 13 virulence genes (chromosomal, prophage, and plasmid) and resistance to 14 antibiotics. A total of 300 samples of mango, 272 of tomato, and 354 of raw chicken were analyzed. The mean of the prevalence (24.9%) and concentration (−0.61 Log MPN/g) of S. enterica in chicken was higher than in mango (1.3%, −1.7 Log MPN/g) and tomato (1.1%, −1.7 Log MPN). Among S. enterica isolates (284), there were 7 different virulotypes, belonging 68.7% of isolates to V2; there was high variability in the presence of mobile genetic elements. The occurrence of specific mobile elements ranged from 81.4% to 11.3% among isolates. Among the isolates, 91.5% were resistant to at least one antibiotic with ampicillin being the most frequent; 54.9% of isolates were multidrug resistant. Data from this study can be used for quantitative microbial risk assessment of S. enterica related to mango, tomato, and raw chicken consumption in the central region of Mexico. Practical Application: Data on the prevalence and concentration of Salmonella enterica obtained in this study can be used to estimate the exposure assessment for the consumption of mango, tomato, and chicken in the central region of Mexico. In addition, the characteristics of the S. enterica isolates could be used to select representative strains for future studies to evaluate the intraspecies variability. [ABSTRACT FROM AUTHOR]

The behavior of Bacillus anthracis Strene spores in sterile raw ground beef is measured at storage temperatures 2 to 70 degree centigrade, which has encompassed both bacterial growth and death. The inability of Bacillus anthracis to grow between 2 and 16 degree centigrade and inactivation at high temperatures might reduce the risk for recommended ground-beef handling and preparation procedures.

The interactions between Escherichia coli O157:H7 and the gastric environment, including comparisons with Shigella spp. are reported. In order to understand the kinetics of Escherichia coli O157:H7 survival in gastric fluid, individual Escherichia coli O157:H7 were suspended in various media mixed at various proportions with simulated human gastric fluid and then, incubated at 37 degree Celsius for up to 4 h.

The dynamics of bacteria community of "Bola de Ocosingo" cheese, a Mexican artisanal raw milk cheese was investigated by high-throughput sequencing (454 pyrosequencing). Dairy samples (raw milk, curd, cheese at 50 and 110 days of ripening) were collected at dry (March-June) and rainy season (August-November) from three producers located in Chiapas, Mexico. In general, raw milk contained high bacterial diversity which was reduced throughout cheese manufacture. However, in two productions an important increase during cheese ripening was observed probably due to cross-contamination. Species such as Streptococcus thermophilus , Lactococcus lactis , Lactobacillus helveticus , L. delbrueckii and L. plantarum from which potential probiotic strains may be obtained, predominated during processing, varying its prevalence from one producer to another. Furthermore, low proportions of Escherichia coli / Shigella flexnerii were detected in almost all processes, however, could not be recovered by traditional methodology, indicating presence of non-cultivable cells. This work provides insights into bacteria communities of Bola de Ocosingo cheese for starter culture development, many of which are reported to provide health related benefits, and the usefulness of high-throughput sequencing to evidence cross-contamination during processing. [ABSTRACT FROM AUTHOR]

Food products move through complex supply chains, which require effective logistics to ensure food safety and to maximize shelf-life. Predictive models offer an efficient means to monitor and manage the safety and quality of perishable foods, however models require environmental data to estimate changes in microbial growth and sensory attributes. Currently, several companies produce Time-Temperature Indicators that react at rates that closely approximate predictive models; these devices are simple and cost-effective for food companies. However, even greater outcomes could be realized using sensors that transfer data to predictive models in real-time. This report describes developments in predictive models designed for supply chain management, as well as advances in environmental sensors. Important innovation can be realized in both supply chain logistics and food safety management by integrating these technologies. [ABSTRACT FROM AUTHOR]

During the processing and handling of commercial blue crab (Callinectes sapidus), Listeria monocytogenes can potentially contaminate cooked meat and grow to hazardous levels. To manage this risk, predictive models are useful tools for designing and implementing preventive controls; however, no model specific for blue crab meat has been published or evaluated. In this study, a cocktail of L. monocytogenes strains was added to pasteurized blue crab meat, which was incubated at storage temperatures from 0 to 35°C. At selected time intervals, L. monocytogenes was enumerated by direct plating onto modified Oxford agar. A primary model was fitted to kinetic data to estimate the lag-phase duration (LPD) and growth rate (GR). Listeria monocytogenes replicated from 0 to 35°C, with GR ranging from 0.004 to 0.518 log CFU/h. Overall, the LPD decreased with increasing temperature, displaying a maximum value of 187 h at 0°C; however, this trend was not consistent. The LPD was not detected at 10°C, and it occurred inconsistently from trial to trial. A secondary GR model (R2 = 0.9892) for pasteurized crab meat was compared with the L. monocytogenes GR in fresh crab meat, demonstrating bias and accuracy factors of 0.98 and 1.36, respectively. The model estimates varied from other published data and models, especially at temperatures ≥5°C, supporting the need for a specific predictive tool for temperature deviations. [ABSTRACT FROM AUTHOR]

Predictive microbiology is a growing field that estimates the behavior of microorganisms in response to environmental conditions. These include the food production and processing operations that occur from farm to [...]

Vibrio parahaemolyticus can accumulate and grow in oysters stored without refrigeration, representing a potential food safety risk. High temperatures during oyster storage can lead to an increase in total viable bacteria counts, decreasing product shelf life. Therefore, a predictive tool that allows the estimation of both V. parahaemolyticus populations and total viable bacteria counts in parallel is needed. A stochastic model was developed to quantitatively assess the populations of V. parahaemolyticus and total viable bacteria in Pacific oysters for six different supply chain scenarios. The stochastic model encompassed operations from oyster farms through consumers and was built using risk analysis software. Probabilistic distributions and predictions for the percentage of Pacific oysters containing V. parahaemolyticus and high levels of viable bacteria at the point of consumption were generated for each simulated scenario. This tool can provide valuable information about V. parahaernolyticus exposure and potential control measures and can help oyster companies and regulatory agencies evaluate the impact of product quality and safety during cold chain management. If coupled with suitable monitoring systems, such models could enable preemptive action to be taken to counteract unfavorable supply chain conditions. [ABSTRACT FROM AUTHOR]

Irradiated ground beef samples (ca. 3-g portions with ca. 25% fat) inoculated with Yersina pestis strain KIM5 (ca. 6.7 log CFU/g) were heated in a circulating water bath stabilized at 48.9, 50, 52.5, 55, 57.5, or 60°C (120, 122, 126.5, 131, 135.5, and 140°E respectively). Average D-values were 192.17, 34.38, 17.11, 3.87, 1.32, and 0.56 min, respectively, with a corresponding z-value of 4.67°C (8.41°F). In related experiments, irradiated ground beef patties (ca. 95 g per patty with ca. 25% fat) were inoculated with Y. pestis strains KIM5 or CDC-A1122 (ca. 6.0 log CFU/g) and cooked on an open-flame gas grill or on a clam-shell type electric grill to internal target temperatures of 48.9, 60, and 71.1°C (120, 140, and 160°E respectively). For patties cooked on the gas grill, strain KIM5 populations decreased from ca. 6.24 to 4.32, 3.51, and -<0.7 log CFU/g at 48.9, 60, and 71.1°C, respectively, and strain CDC-A1122 populations decreased to 3.46 log CFU/g at 48.9°C and to ≤0.7 log CFU/g at both 60 and 71.1°C. For patties cooked on the clam-shell grill, strain KIM5 populations decreased from ca. 5.96 to 2.53 log CFU/g at 48.9°C and to ≤0.7 log CFU/g at 60 or 71.1°C, and strain CDC-A1122 populations decreased from ca. 5.98 to ≤0.7 log CFU/g at all three cooking temperatures. These data confirm that cooking ground beef on an open-flame gas grill or on a clam-shell type electric grill to the temperatures and times recommended by the U.S. Department of Agriculture and the U.S. Food and Drug Administration Food Code, appreciably lessens the likelihood, severity, and/or magnitude of consumer illness if the ground beef were purposefully contaminated even with relatively high levels of Y. pestis. [ABSTRACT FROM AUTHOR]

From 2003 to 2004, we studied the impact of environmental influences on the microbiological quality of a hydroponic tomato farm. The presence of Salmonella was investigated on 906 samples of tomatoes and 714 environmental samples. The farm comprised 14 greenhouses and a technologically advanced packinghouse, and operated under a sanitary agricultural practices plan. The objective of the present study was to determine the operating sources of contamination. During the course of the study, two independent natural events affected the farm. In 2003, water runoff entered some of the greenhouses. A year later, wild animals (opossums, mice, and sparrows) gained entry into several of the greenhouses. Salmonella and Escherichia coli were found in samples of tomatoes, water puddles, soil, shoes, and the feces of local wild and farm animals. Salmonella Montevideo, Salmonella Newport, and strains of the F serogroup were isolated from tomatoes. Almost all of the Salmonella Newport strains were isolated from samples collected during or immediately after the flood. Analysis by pulsed-field gel electrophoresis revealed that some Salmonella Montevideo isolates from tomatoes, opossums, and mice displayed identical XbaI or AvrII patterns, suggesting that these wild animals represented one source of contamination. F serogroup strains were found mostly on samples of goat feces and personnel shoes when standard working practices were in place. Shoes were found to be an important vehicle for dissemination of Salmonella into the greenhouses. The level of protection provided by hydroponic greenhouses does not exclude the eventuality that enteric pathogenic bacteria can gain access through various avenues. [ABSTRACT FROM AUTHOR]

The increasing demand of ready-to-eat poultry products has led to serious concerns over product safety, and more emphasis has been placed on thorough cooking of products. In this study, processing conditions and thermal inactivation of Listeria innocua in chicken breast meats were evaluated during convection cooking in a pilot-plant scale air-steam impingement oven. A predictive model was developed by integrating heat and mass transfer models with a pathogen kinetics model to predict temperature, water content, product yield, and bacterial inactivation during air-steam impingement cooking. Skinless boneless chicken breasts were cooked at oven air temperatures of 177 and 200°C for 2 to 10 min at a humidity of 70 to 75% (moisture by volume) and an air velocity of 1 m/s at the exit of the nozzles. The reduction in Listeria in chicken breasts after 2 to 5 min of cooking was from 0.3 to 1.4 log CFU/g and from 0.8 to 1.8 log CFU/g at 177 and 200°C, respectively. After cooking for 10 min at both temperatures, no survivors were detected in any of the cooked chicken breasts from an initial bacterial concentration of 106 CFU/g. The standard errors of prediction for the endpoint center temperatures after 2 to 10 min of cooking were 2.8 and 3.0°C for air temperatures of 177 and 200°C, respectively. At 177 and 200°C, the median relative errors of prediction for water content were 2.5 and 3.7% and those for product yield were 5.4 and 8.4%, respectively. The developed model can be used as a tool to assist in evaluating thermal processing schedules for poultry products cooked in an air-steam impingement oven. [ABSTRACT FROM AUTHOR]

The influences of the relative humidity (RH) and storage temperature on the colonization of tomato surfaces by Salmonella Montevideo were studied. Red, ripe tomatoes (Lycopersicon esculentum) were spot inoculated in three separate trials with 100 µl (approximately 106 CFU) of Salmonella Montevideo and stored for 90 min at 22°C under 97% RH to facilitate attachment of cells to the blossom end of tomato surfaces. Following this attachment step, tomatoes were washed to remove loosely adhered cells and then stored at 22 or 30°C for up to 10 days under RH of 60, 75, 85, or 97%. At 0, 0.4, 1, 4, 7, and 10 days of storage, three tomatoes were individually hand massaged in 50 ml of 0.1% peptone water and the washes were separately analyzed to enumerate populations of Salmonella Montevideo. The number of Salmonella Montevideo cells attached after 90 min at 22°C was 3.8 log CFU per tomato; this level was determined to be the initial colonizing population. After 10 days of storage at 30°C, the Salmonella Montevideo population increased to 0.7, 1.0, 1.2, and 2.2 log CFU per tomato at 60, 75, 85, and 97% RH, respectively. A similar trend was observed at 22°C, although populations were lower than at 30°C. Scanning electron micrographs of tomato cuticles after storage revealed a well-defined biofilm containing bacteria. These findings reinforce the importance of maintaining stored tomatoes at temperatures that do not support growth of pathogenic bacteria and demonstrate the growth-promoting effects of high humidity. [ABSTRACT FROM AUTHOR]

A five-strain cocktail of Listeria monocytogenes (104 cfu/mL ) was inoculated onto individual vacuum-packaged slices (ca. 50 g each) of a commercial, Hispanic-style cheese, that being Queso Blanco. Growth was determined at appropriate intervals during storage at 5, 10, 15, 20 and 25C. In general, as the incubation temperature increased, a shorter lag phase duration (LPD) and a faster growth rate (GR) were observed. The LPD values at 5, 10, 15, 20 and 25C were 65.3, 19.9, 2.1, 8.4 and 11.4 h, respectively. The GR values were 0.011, 0.036, 0.061, 0.090 and 0.099 log cfu/h at 5, 10, 15, 20 and 25C, respectively. There were no statistical differences in LPD at 10, 15, 20 and 25C. However, the LPD during growth at 5C was statistically (P ≤ 0.05) longer than at all other temperatures. The GR values at 20 and 25C were not significantly different from each other, whereas the GR values at 5, 10 and 15C were significantly different from each other as well as from the GR at 20 and 25C (P ≤ 0.05). The maximum population density (MPD) showed relatively little variation over the range of storage temperatures tested, with an average of 8.38 log cfu/g (SD = 0.33). The results of this study indicate that not even the lowest trial temperature of 5C prevented growth over time of the inoculated L. monocytogenes on this sliced product, and that proper storage and handling procedures are required to prevent the bacterium from contaminating the product and/or to control its growth. [ABSTRACT FROM AUTHOR]

Risk studies have identified cross-contamination during beef fabrication as a knowledge gap, particularly as to how and at what levels Escherichia coli O157:H7 transfers among meat and cutting board (or equipment) surfaces. The objectives of this study were to determine and model transfer coefficients (TCs) between E. coli O157:H7 on beef tissue and high-density polyethylene (HDPE) cutting board surfaces. Four different transfer scenarios were evaluated: (i) HDPE board to agar, (ii) beef tissue to agar, (iii) HDPE board to beef tissue to agar, and (iv) beef tissue to HDPE board to agar. Also, the following factors were studied for each transfer scenario: two HDPE surface roughness levels (rough and smooth), two beef tissues (fat and fascia), and two conditions of the initial beef tissue inoculation with E. coli O157:H7 (wet and dry surfaces), for a total of 24 treatments. The TCs were calculated as a function of the plated inoculum and of the cells recovered from the first contact. When the treatments were compared, all of the variables evaluated interacted significantly in determining the TC. An overall TC-per-treatment model did not adequately represent the reduction of the cells on the original surface after each contact and the interaction of the factors studied. However, an exponential model was developed that explained the experimental data for all treatments and represented the recontamination of the surfaces with E. coli O157:H7. The parameters for the exponential model for cross-contamination with E. coli O157:H7 between beef tissue and HDPE surfaces were determined, allowing for the use of the resulting model in quantitative microbial risk assessment. [ABSTRACT FROM AUTHOR]

The robustness of a microbial growth model must be assessed before the model can be applied to new food matrices; therefore, a methodology for quantifying robustness was developed. A robustness index (RI) was computed as the ratio of the standard error of prediction to the standard error of calibration for a given model, where the standard error of calibration was defined as the root mean square error of the growth model against the data (log CFU per gram versus time) used to parameterize the model and the standard error of prediction was defined as the root mean square error of the model against an independent data set. This technique was used to evaluate the robustness of a broth-based model for aerobic growth of Escherichia coli O157:H7 (in the U.S. Department of Agriculture Agricultural Research Service Pathogen Modeling Program) in predicting growth in ground beef under different conditions. Comparison against previously published data (132 data sets with 1,178 total data points) from experiments in ground beef at various experimental conditions (4.8 to 45°C and pH 5.5 to 5.9) yielded RI values ranging from 0.11 to 2.99. The estimated overall RI was 1.13. At temperatures between 15 and 40°C, the RI was close to and smaller than 1, indicating that the growth model is relatively robust in that temperature range. However, the RI also was related (P < 0.05) to temperature. By quantifying the predictive accuracy relative to the expected accuracy, the RI could be a useful tool for comparing various models under different conditions. [ABSTRACT FROM AUTHOR]

In eight separate trials, four groups of raw ground beef samples were inoculated with 0.04 to 0.3 CFU/g of Salmonella Typhimurium (DT 104). Each group consisted of four 25-g samples (three inoculated and one uninoculated). After inoculation, these samples were shipped by overnight courier in shipping containers with ice packs from the U.S. Department of Agriculture (USDA), Eastern Regional Research Center, in Wyndmoor, Pa., to the U.S. Food Safety and Inspection Service (FSIS), Eastern Laboratory, in Athens, Ga. A total of 128 samples (32 in each of four groups) were shipped. A temperature data logger was placed inside each shipping container to record the temperature during shipping and storage. The first group of ground beef samples was analyzed within approximately I h of arrival. The second group of samples was left in the original containers, with a gel ice pack, for 24 h before processing. The third and fourth groups of samples were removed from the original shipping containers and stored at room temperature (21 ± 2°C) for 6 h and then in a refrigerator at 4 ± 2°C for 24 and 48 h, respectively, before analysis. The samples were analyzed for the presence of Salmonella according to the USDA/FSIS Microbiological Laboratory Guidebook, chapter 4.02. There was no significant difference in the presence and levels of Salmonella in ground beef among the four test groups. These data show that it is acceptable to process the late-arriving ground beef samples for the detection of Salmonella if they are kept in a refrigerator (4 ± 2°C) for 24 to 48 h or when the shipments arrive late (24 h in the container with ice pack). [ABSTRACT FROM AUTHOR]

Abstract: Seafood salad has been identified as a ready-to-eat food with a relatively high incidence of contamination by Listeria monocytogenes; however, little is known about the behavior of this pathogen in seafood salad as a function of product pH and storage temperature. To produce data towards the development of a predictive growth model, a 6-strain cocktail of L. monocytogenes was inoculated onto the surface of a shrimp-crabmeat product, mixed with mayonnaise that was previously adjusted with NaOH to pH 3.7, 4.0, 4.4, 4.7 or 5.1, and then stored at 4°, 8° or 12 °C under both aerobic and vacuum conditions. At each storage temperature, L. monocytogenes was able to grow in the seafood salad under both aerobic and vacuum conditions. The slowest growth of L. monocytogenes was observed in seafood salad with a mayonnaise pH of 3.7 and a storage temperature of 4 °C under vacuum condition. In salad with the same mayonnaise pH, the growth rate (GR, log10 cfu/h) of L. monocytogenes increased as a function of storage temperature. At the same storage temperature, the lag phase duration (LPD, h) of L. monocytogenes decreased as mayonnaise pH increased. At the same mayonnaise pH and temperature, LPD of L. monocytogenes was greater under aerobic than under vacuum conditions. Regression analyses indicated that mayonnaise pH is the main effector on the LPD of L. monocytogenes in seafood salad, and storage temperature was the main effector on the GR. Secondary models that describe LPD and GR of L. monocytogenes in seafood salad as a function of mayonnaise pH and storage temperature were produced. [Copyright &y& Elsevier]

Abstract: Escherichia coli O157:H7 can contaminate raw ground beef and cause serious human foodborne illness. Previous reports describe the behavior of E. coli O157:H7 in ground beef under different storage conditions; however, models are lacking for the pathogen''s behavior in raw ground beef stored over a broad range of temperature. Using sterile irradiated raw ground beef, the behavioral kinetics of 10 individual E. coli O157:H7 strains and/or a 5- or 10-strain cocktail were measured at storage temperatures from 5° to 46 °C. Growth occurred from 6 to 45 °C. Although lag phase duration (LPD) decreased from 10.5 to 45 °C, no lag phase was observed at 6, 8, or 10 °C. The specific growth rate (SGR) increased from 6 to 42 °C then declined up to 45 °C. In contrast to these profiles, the maximum population density (MPD) declined with increasing temperature, from approximately 9.7 to 8.2 log cfu/g. Bias (B f) and accuracy (A f) factors for an E. coli O157:H7 broth-based aerobic growth model (10 to 42 °C) applied to the observations in ground beef were 1.05, 2.70, 1.00 and 1.29, 2.87, 1.03, for SGR, LPD and MPD, respectively. New secondary models increased the accuracy of predictions (5 to 45 °C), with B f and A f for SGR, LPD, and MPD of 1.00, 1.06, and 1.00 and 1.14, 1.33, and 1.02, respectively. These new models offer improved tools for designing and implementing food safety systems and assessing the impact of E. coli O157:H7 disease. [Copyright &y& Elsevier]

Several outbreaks of listeriosis have been associated with contamination of wieners and other ready-to-eat meat products. In this study, we addressed the question of whether emulsification in, or growth on, wieners triggers a response in the listerial cells that makes them more virulent or protects them against the harsh environment of the gastrointestinal tract in mice. Our results indicate that Listeria monocytogenes Scott A grows poorly, if at all, in one brand of commercially prepared wieners inoculated with 5 × 10³ to 5 × 106 CFU per package and incubated at 15°C. Neither L. monocytogenes Scott A emulsified in a slurry of homogenized wieners nor recovered from wiener package fluid after a 7-day incubation at 15°C were more virulent when inoculated into the stomachs of A/J mice than L. monocytogenes Scott A grown in brain heart infusion broth. These findings suggest that the ability of L. monocytogenes Scott A to cause systemic infection following introduction into the gastrointestinal tract was not improved by incubation with wieners or suspension in a meat matrix. [ABSTRACT FROM AUTHOR]

The advancement of predictive microbiology relies on available data that describe the behavior of microorganisms in different environmental matrices. For such information to be useful to the predictive microbiology research community, data must be organized in a manner that permits efficient access and data retrieval. Here, we describe a database protocol that encompasses observations of bacterial responses to food environments, resulting in a database (ComBase) for predictive microbiology purposes. The data included in ComBase were obtained from cooperating research institutes and from the literature and are publicly available via the Internet. [ABSTRACT FROM AUTHOR]

Tests for fecal (thermotolerant) and total coliforms applied to drinking and recreational waters are convenient but not necessarily accurate indicators of the presence of pathogens originating from the intestines of warm-blooded animals. These tests may be inappropriate for analyses of complex industrial effluents such as pulp mill effluents, which may contain bacteria that yield a positive result in fecal and total coliform tests, even though no fecal source exists. Consequently, other more specific indicators for the presence of potential pathogens are needed. In cases where the sources of, or validity of, fecal responses are problematic, the direct examination of pathogens, and/or the use of new source-tracking techniques, such as DNA fingerprinting, may be better approaches for protecting public health. [ABSTRACT FROM AUTHOR]

Focuses on a study which reported the characterization of a clinical Vibrio cholerae O319 isolate from a patient in Mexico diagnosed with gastrointestinal illness. Bacterial isolates obtained from stool samples of the patient; Standard tests used for the biochemical characterization of the isolates; Findings on the isolates.

Vibriospp. cause frank and opportunistic infections of humans through exposure to seafood and seawater. Due to their natural occurrence in coastal environments, traditional indicator organisms, such asE. coli, do not predict their presence. This problem has complicated public health initiatives aimed at reducing the impact of illnesses fromVibriospp. In the U.S.,V. vulnificushas received extensive study due to the severity of its disease in humans. Its numbers increase with warmer summer temperature, and decline to nondetectable levels in colder winter months. In environments with salinities greater than 20 ppt,V. vulnificusnumbers decline to levels that do not pose human health risks. A similar response to temperature has been observed for pathogenic strains ofV. parahaemolyticus, where recent outbreaks of illness have been associated with El Niño weather conditions. In addition, temperature-induced plankton blooms have been linked to epidemic cholera in certain geographical regions of the world. New research shows that seawater temperature and salinity can be used to develop mathematical models ofV. vulnificusincidence in coastal environments. Similar efforts might be extended to otherVibriospp. to develop indicators that predict human health risk, as well as ecosystem integrity. [ABSTRACT FROM PUBLISHER]

Food regulatory authorities permit the use of Time as Public Health Control (TPHC) for handling foods that potentially support the growth of pathogenic bacteria. Considering the widespread use of TPHC in food service operations, few reports quantitatively describe potential pathogen growth when these protocols are implemented. A worst-case growth rate model was built from the highest growth rates predicted by ComBase broth-based models for six pathogens. A separate worst-case growth model was constructed from growth rates in ComBase database records. The maximum estimated pathogen growth in 4 h, assuming no lag phase, ranged from 0.006 log CFU at 5 °C to 6.16 log CFU at 44 °C, with 3.1 log CFU at 25 °C. In addition, pathogen growth when implementing TPHC could exceed the 1- and 3-log limits recommended for food challenge tests. The use of predictive models in development of TPHC criteria may provide more fail-safe strategies for managing microbial hazards in potentially hazardous food. This strategy could also reduce food waste and promote the use of temperature sensors in food supply chains. • 'Time as Public Health Control' can produce 0.006 to 6.16 log CFU pathogen growth. • Pathogen growth can exceed 1- and 3-log CFU recommended pathogenic species limits. • Predictive models can help design safer time-temperature food handling protocols. [ABSTRACT FROM AUTHOR]

Reports on the use of restriction fragment length polymorphism (RFLP) and biotype profiles for environmental and clinical isolates of Vibrio vulnificus using pulsed-field gel electrophoresis (PFGE). PFGE binary metric plot of average similarity linkage of isolates; Percentages of clinical and environmental isolates at selected similarity indices.

Australian regulations for microbiological testing of carcasses specify a number of incubation temperatures and media for meat processed at both domestic and export establishments. Accordingly, the effect of incubation temperature and media on aerobic plate counts of samples from beef and sheep carcasses was investigated. For both species, aerobic plate counts on Petrifilm incubated at 35°C were significantly lower than those counts on Petrifilm and pour plates incubated at 25 and 30°C, reflecting the inability of many psychrotrophs to grow at 35°C. When samples were taken from carcasses that had been stored in abattoir chillers for periods between 16 h and 5 days, difference between counts at 35°C versus those incubated at 25 and 30°C became greater as the period of refrigerated storage increased. For export beef carcasses, the effect of this difference is minimal, since the vast majority of counts incubated at 35°C are done on carcasses that have been chilled for less than 24 h and will not have a large proportion of psychrotrophs. [ABSTRACT FROM AUTHOR]

The bacterial diversity and structure of Poro cheese, an artisanal food, was analysed by high-throughput sequencing (454 pyrosequencing) in order to gain insight about changes in bacterial communities associated with the cheese-making process. Dairy samples consisting of milk, fermented whey, curd and ripened cheese (during 7 and 60 d) were collected from three manufacturers located in the state of Tabasco, México during dry (March-June) and rainy (August-November) seasons. Independently of producer and season, raw milk samples displayed the highest diversity in bacterial communities. In raw milk, genera found were Macrococcus, Staphylococcus, Enterococcus, Streptococcus, Lactobacillus and Enhydrobacter. Diversity in whey, curd and cheese was lower, principally containing Streptococcus and Lactobacillus; however, bacteria such as Staphylococcus, Acinetobacter, Chryseobacterium, Bacillus, Sediminibacter, Lactococcus and Enterococcus were occasionally present. After curdling step, the most dominant and abundant species were Streptococcus thermophilus and Lactobacillus delbrueckii. [ABSTRACT FROM AUTHOR]

Abstract: The potential impact of post-pasteurisation contamination of liquid egg products with the multi-antibiotic resistant pathogen Salmonella enterica serotype Typhimurium definitive type 104 (DT104) was assessed by determining the viability of this bacterium in whole egg, albumen and 10% w/w sugared and salted yolk incubated at 4–42 °C. Results indicated that populations of S. Typhimurium DT104 were slowly inactivated in all four products when stored at 4 °C. However, based on the typical shelf-lives of cold-stored liquid egg, less than 0.6 log-kill would be achieved in those products prior to their use. Incubation at temperatures pertaining to abuse situations (10, 15, 20 and 25 °C) revealed an increasing potential for growth of S. Typhimurium DT104 in whole egg, albumen and sugared yolk, as indicated by trends in growth rate, lag duration and maximum population density. At even higher temperatures (30, 37 and 42 °C), growth rates of S. Typhimurium DT104 in whole egg and sugared yolk continued to increase. The same was true for S. Typhimurium DT104 in albumen except that growth was not observed at 42 °C and instead populations were inactivated within 30 h. At no temperature tested was S. Typhimurium DT104 able to grow in salted yolk. The influence of these growth and inactivation patterns on the risk of salmonellosis in relation to product type and storage temperature is discussed. [Copyright &y& Elsevier]

Reports on a study which defined the components of the enrichment broth which improved and detected the growth of the Vibrio vulnificus. In-depth look at the alkaline peptone broth (APB); Evaluation of the impact of the APB on the components of the Vibrio vulnificus growth; Methodology used to conduct the study; Results of the study.

Predictive models offer efficient means to manage the quality and safety of highly perishable seafood. Salmon is an increasingly popular seafood, and relies on well managed domestic and international supply chains to minimize growth of spoilage and pathogenic bacteria. While the literature describes predictive models for smoked and modified atmosphere packaged salmon, there are no reported models for spoilage bacteria and Listeria monocytogenes on head-on and gutted (HOG) aerobically-stored Atlantic salmon. Predictive models were developed for microbial and sensorial degradation of HOG Atlantic salmon stored at 0–15 °C until the end of shelf-life. Total Viable Count (TVC) and Pseudomonas spp. had similar growth rates at 0, 5 and 10 °C, but TVC rate was higher at 15 °C. L. monocytogenes growth rate at 0 °C was 0.004 log 10 cfu/h, and showed a log-linear increase (R 2 = 0.99) to 0.079 log 10 cfu/h at 15 °C. Sensory Quality Index (QI) scores were 2.4, 4.5, and 7.2 times greater at 5, 10 and 15 °C, respectively, compared to 0 °C. QI and TVC rates had a relatively strong relationship at 5 (R 2 = 0.87), 10 (R 2 = 0.80) and 15 °C (R 2 = 0.78), compared to 0 °C (R 2 = 0.50). These models are potential tools to manage the safety and quality of HOG Atlantic salmon in supply chains. [ABSTRACT FROM AUTHOR]

Primary and secondary models were developed for quantitatively characterizing the survival of Listeria monocytogenes in soy-sauce based acidified Asian style products that do not undergo a thermal treatment. The objective of this study was to quantify the effect of food matrix properties on L. monocytogenes ' survival in soy sauce-based products. This quantification enables a product-specific estimation of 5-log reduction time to ensure a safe processing and management operation, to ultimately facilitate a science-based, safety-oriented product development process. A central composite design with four independent variables (pH, soy sauce, added NaCl and soluble solids) with five levels was used to plan the challenge studies on different formulations. To model microbial survival over time, different non-linear primary models were fit to the data obtained from challenge studies. The best-fit model was selected based on a series of statistical goodness-of-fit measures. Kinetic parameters estimated from the best-fit primary models were fit to response surface equations using second order polynomial regression. The best-fit primary model representative of the product formulations was a modified Weibull model. The natural logarithm of the scale parameter (δ , in h) was used as the response variable for the secondary model. This resulted in acceptable fitting compared to the observed values with R 2 values of 0.95 and RMSE of 0.7 h. External validity of model predictions was conducted by comparing them to 5-log reduction times observed in independent challenge tests using different product formulations. Results indicated an acceptable validation with R 2 = 0.81 and RMSE = 35 h. The present study provides quantitative tools specific for cold-fill-hold soy sauce-based products to enhance microbial safety management plans and product development. • Primary and secondary predictive models developed for L. monocytogenes • Specific predictive models needed for complex food matrices • Research & development and food safety applications in industry are targeted. • Soy sauce, pH, NaCl and Brix are significant factors for L. monocytogenes survival. [ABSTRACT FROM AUTHOR]

Abstract: The mechanisms of bacterial attachment to meat tissues need to be understood to enhance meat safety interventions. However, little is known about attachment of foodborne pathogens to meat muscle cells. In this study, attachment of six Escherichia coli and two Salmonella strains to primary bovine muscle cells and a cultured muscle cell line, C2C12, was measured, including the effect of temperature. At 37°C, all but one strain (EC623) attached to C2C12 cells, whereas only five of eight strains (M23Sr, H10407, EC473, Sal1729a and Sal691) attached to primary cells. At 10°C, two strains (H10407 and EC473) attached to C2C12 cells, compared to four strains (M23Sr, EC614, H10407 and Sal1729a) of primary cells. Comparing all strains at both temperatures, EC614 displayed the highest CFU per C2C12 cell (4.60±2.02CFU/muscle cell at 37°C), whereas greater numbers of M23Sr attached per primary cell (51.88±39.43CFU/muscle cell at 37°C). This study indicates that primary bovine muscle cells may provide a more relevant model system to study bacterial attachment to beef carcasses compared to cell lines such as C2C12. [Copyright &y& Elsevier]

Abstract: Seven blue crab processing plants were sampled to determine the prevalence and sources of Listeria spp. and Listeria monocytogenes for two years (2006–2007). A total of 488 raw crabs, 624 cooked crab meat (crab meat) and 624 environmental samples were tested by standard methods. Presumptive Listeria spp. were isolated from 19.5% of raw crabs, 10.8% of crab meat, and 69.5% of environmental samples. L. monocytogenes was isolated from 4.5% of raw crabs, 0.2% of crab meat, and 2.1% of environmental samples. Ninety-seven percent of the isolates were resistant to at least one of the ten antibiotics tested. Eight different serotypes were found among 76 L. monocytogenes isolates tested with the most common being 4b, 1/2b and 1/2a. Automated EcoRI ribotyping differentiated 11 ribotypes among the 106 L. monocytogenes isolates. Based on ribotyping analysis, the distribution of the ribotypes in each processing plant had a unique contamination pattern. A total of 92 ApaI and 88 AscI pulsotypes among the 106 L. monocytogenes isolates were found and distinct pulsotypes were observed in raw crab, crab meat and environmental samples. Ribotypes and serotypes recovered from crab processing plants included subtypes that have been associated with listeriosis cases in other food outbreaks. Our findings suggest that molecular methods may provide critical information about sources of L. monocytogenes in crab processing plants and will augment efforts to improve food safety control strategies such as targeting specific sources of contamination and use of aggressive detergents prior to sanitizing. [Copyright &y& Elsevier]

Abstract: Meat surfaces are contaminated with bacteria during slaughter and processing. Understanding bacterial attachment properties to specific structures of meat could result in more targeted interventions to improve its safety and quality. However, the influence of temperatures relevant to abattoir environments on bacterial attachment to specific meat structures is not known. In this study, the effect of temperature and protein concentration on attachment of 10 Escherichia coli and seven Salmonella strains to extracellular matrix (ECM) proteins (collagen I, fibronectin, collagen IV and laminin) was measured using crystal violet stain and epifluorescence microscopy assays. By crystal violet assay, only five of 17 strains showed significant attachment to any ECM protein and only one strain attached to all proteins. Strains that attached at all tested temperatures (4, 25, 37°C) were E. coli M23Sr and M23 (collagen I); E. coli M23Sr (fibronectin); E. coli M23Sr, O157:H12 and M23, (collagen IV); and E. coli M23Sr, O157:H12, O78:K80:H1, O26:H11 and M23 (laminin). A higher proportion of strains attached to basement membrane proteins (laminin and collagen IV) than to interstitial proteins (collagen I and fibronectin). Highest attachment levels occurred at 4°C for collagen I and at 25°C for the other three proteins. Generally, the attachment levels of Salmonella strains to all ECM proteins were lower than for E. coli. No significant effect was found for concentration of collagen I, fibronectin and collagen IV, but was for higher laminin concentration. A strong positive correlation was found between results of both the crystal violet and epifluorescent methods (r≥0.905, p <0.05). This study demonstrated that attachment properties to ECM proteins displayed distinct variation among strains, that temperature highly influenced attachment and that protein concentration had a minor effect. [Copyright &y& Elsevier]

Postharvest growth of Vibrio vulnificus in oysters can increase risk of human infection. Unfortunately, limited information is available regarding V. vulnificus growth and survival patterns over a wide range of storage temperatures in oysters harvested from different estuaries and in different oyster species. In this study, we developed a predictive model for V. vulnificus growth in Eastern oysters (Crassostrea virginica) harvested from Chesapeake Bay, MD, over a temperature range of 5 to 30°C and then validated the model against V. vulnificus growth rates (GRs) in Eastern and Asian oysters (Crassostrea ariakensis) harvested from Mobile Bay, AL, and Chesapeake Bay, VA, respectively. In the model development studies, V. vulnificus was slowly inactivated at 5 and 10°C with average GRs of −0.0045 and −0.0043 log most probable number (MPN)/h, respectively. Estimated average growth rates at 15, 20, 25, and 30°C were 0.022, 0.042, 0.087, and 0.093 log MPN/h, respectively. With respect to Eastern oysters, bias (Bf) and accuracy (Af) factors for model-dependent and -independent data were 1.02 and 1.25 and 1.67 and 1.98, respectively. For Asian oysters, Bf and Af were 0.29 and 3.40. Residual variations in growth rate about the fitted model were not explained by season, region, water temperature, or salinity at harvest. Growth rate estimates for Chesapeake Bay and Mobile Bay oysters stored at 25 and 30°C showed relatively high variability and were lower than Food and Agricultural Organization (FAO)/WHO V. vulnificus quantitative risk assessment model predictions. The model provides an improved tool for designing and implementing food safety plans that minimize the risk associated with V. vulnificus in oysters. [ABSTRACT FROM AUTHOR]

Vibrio parahaemolyticus is an indigenous bacterium of marine environments. It accumulates in oysters and may reach levels that cause human illness when postharvest temperatures are not properly controlled and oysters are consumed raw or undercooked. Predictive models were produced by injecting Pacific oysters (Crassostrea gigas) with a cocktail of V. parahaemolyticus strains, measuring viability rates at storage temperatures from 3.6 to 30.4°C, and fitting the data to a model to obtain parameter estimates. The models were evaluated with Pacific and Sydney Rock oysters (Saccostrea glomerata) containing natural populations of V. parahaemolyticus. V. parahaemolyticus viability was measured by direct plating samples on thiosulfate-citratebile salts-sucrose (TCBS) agar for injected oysters and by most probable number (MPN)-PCR for oysters containing natural populations. In parallel, total viable bacterial counts (TVC) were measured by direct plating on marine agar. Growth/inactivation rates for V. parahaemolyticus were -0.006, -0.004, -0.005, -0.003, 0.030, 0.075, 0.095, and 0.282 log10 CFU/h at 3.6, 6.2, 9.6, 12.6, 18.4, 20.0, 25.7, and 30.4°C, respectively. The growth rates for TVC were 0.015, 0.023, 0.016, 0.048, 0.055, 0.071, 0.133, and 0.135 log10 CFU/h at 3.6, 6.2, 9.3, 14.9, 18.4, 20.0, 25.7, and 30.4°C, respectively. Square root and Arrhenius-type secondary models were generated for V. parahaemolyticus growth and inactivation kinetic data, respectively. A square root model was produced for TVC growth. Evaluation studies showed that predictive growth for V. parahaemolyticus and TVC were "fail safe." The models can assist oyster companies and regulators in implementing management strategies to minimize V. parahaemolyticus risk and enhancing product quality in supply chains. [ABSTRACT FROM AUTHOR]

Abstract: In this study, we investigated the role of lysozyme on the viability of Bacillus cereus, Bacillus subtilis, Bacillus pumilus and Bacillus anthracis (Sterne) in egg white (EW), ground beef and milk. At 35 °C in EW, growth rates (GR) for B. cereus, B. subtilis, B. pumilus and B. anthracis were 0.005, −0.018, −0.028 and −0.029 OD600/h, respectively. Heat-treating EW at 55 and 60 °C reduced the inactivating effect of EW by 3.1 and 10.5-fold, respectively. Addition of lysozyme (2 mg/ml) to 60 °C-treated EW increased the inactivation rate 5.76-fold, indicating involvement of lysozyme in B. anthracis inactivation. B. anthracis inactivation was influenced by pH, as shown by a progressive increase in inactivation rate from 0.25 to −4.42 logs CFU/h over a pH range of 6.0–8.5. Adding 2 mg/ml lysozyme to milk and ground beef also suppressed the growth of B. anthracis 3.3 and 6.5-fold, respectively. These data indicate that lysozyme, as a natural component of EW or potential additive in other foods, could reduce biothreat risks presented by bioterror agents. [Copyright &y& Elsevier]

Abstract: The thermal stability of heat-shocked and non-heat-shocked spores of the virulence-attenuated Sterne strain of Bacillus anthracis was evaluated at select temperatures in irradiated, raw ground beef (25% fat) heated in a water bath or cooked using two different commercial grills. For the former, 3-g portions of inoculated ground beef were packaged in bags that were completely immersed in a temperature-controlled circulating water bath held at 65°C (149°F), 70°C (158°F), 75°(167°F), and 80°C (176°F) for a predetermined length of time. For the latter, formed ground beef patties (95-g each) were inoculated with spore stock A or B of the Sterne strain and then cooked on a commercial open-flame gas grill or on a commercial clamshell electric grill to achieve target internal temperatures of either 71.1°C (160°F), 82.2°C (180°F), or 93.3°C (200°F). Cooking ground beef patties on commercial grills, resulted in reductions of ca. 0.8 to 3.5log10 CFU/g for spore stocks A and B of B. anthracis Sterne after heating to 71.1°C (160°F), 82.2°C (180°F), or 93.3°C (200°F) on either the open-flame gas grill which required ca. 9.6min to reach the target internal temperatures or on the clamshell electric grill which required ca. 4.0min to reach the target internal temperatures. In comparison, our data using a water bath system and heating at 65° to 80°C predict nearly 4log reductions in spore levels for short times, ~½min, depending possibly on the temperature. Thus, our data suggest that models based on heating ground beef in a water bath is not a good predictor of reductions of levels of spores of B. anthracis Sterne strain that would be obtained when cooking ground beef patties on commercial grills under conditions that may be typically used by consumers and/or retail establishments. Nevertheless, our data validated that cooking ground beef patties on a commercial grill at a temperature considered to be “well-done” and a temperature (71.1°C;160°F) recommended by the USDA/FSIS, is effective at killing spores of B. anthracis Sterne. Industrial relevance: Heating ground beef in a water bath or cooking ground beef patties on commercial grills under conditions simulating those that are used by consumers and/or that occur in retail food service establishments is effective at killing spores of B. anthracis Sterne. [Copyright &y& Elsevier]