Simulation modelling and risk assessment as tools to identify the impact of climate change on microbiological food safety – The case study of fresh produce supply chain

The current quality assurance and control tools and methods to prevent and/or to control microbiological risks associated with fresh produce are challenged due to the following pressures upon the food supply chain, i.e. changing consumption patterns, globalization and climate change. It demonstrates the need for scientific research and development of new and/or improved tools, techniques and practices to adapt the current risk management systems. In this paper, a conceptual research approach is presented to analyse the complexity of the climate change and globalization challenge on the fresh produce supply chain taken as a case study. The factors which affect the vulnerability of the fresh produce chain demand a multidisciplinary research approach. The proposed knowledge-based modelling system is believed to be a most appropriate way to identify problems and to offer solutions to monitor and prevent microbiological food safety risks during all phases of food production and supply. To explore the potential impact of climate change and globalization, baseline information can be obtained by surveillance and performance measurement of implemented food safety management systems. Simulation of climate change scenarios and the logistic chain of fresh produce, along with mathematical models to optimize packaging technology to maintain quality and safety of fresh produce are tools to provide insights in the complex dynamic ecosystem. They are the basis for elaboration of risk assessment studies to scientifically support management options and decisions to new microbiological threats related to globalization and climate change in the fresh produce supply chain. This research concept as such will contribute to develop strategies in order to guarantee the (microbiological) food safety of fresh produce on the long term.     

Characterization of Escherichia coli from raw poultry in Belgium and impact on the detection of Campylobacter jejuni using Bolton broth

A comparative study examining Bolton broth and Preston broth for enrichment and reliable detection of Campylobacter jejuni (both healthy and freeze stressed cells) was performed. Tested as pure cultures, Bolton broth enabled faster resuscitation and growth of C. jejuni compared to Preston broth. When C. jejuni was co-incubated with extended-spectrum-beta-lactamase (ESBL) producing Escherichia coli isolated from Belgian poultry meat preparations, the latter dominated in the Bolton enrichment broth and crowded the mCCDA plates. This resulted in the inability to recover C. jejuni by ISO 10272-1:2006 standard method. Preston broth did not support the growth of the ESBL E. coli isolates, but showed longer detection time of C. jejuni compared to Bolton broth. The use of the same antibiotic (sodium cefoperazone) in Bolton broth and in mCCDA plates may explain the problems encountered for detection of C. jejuni, as high numbers of ESBL E. coli present after enrichment in Bolton broth, also caused overgrowth and masked the few C. jejuni colonies present on the mCCDA plates. The use of Campylobacter spp. specific real-time PCR circumvented these problems and enabled rapid detection of the pathogen after 24 h enrichment in both Bolton and Preston broth, for both healthy and freeze stressed cells.     

Review: norovirus prevalence in Belgian, Canadian and French fresh produce: a threat to human health?

Foodborne viruses, especially noroviruses (NoV), are increasingly reported as the cause of foodborne outbreaks. NoV outbreaks have been reported linked to fresh soft red fruits and leafy greens. Belgium, Canada and France were the first countries to provide data about the prevalence of NoV on fresh produce. In total, 867 samples of leafy greens, 180 samples of fresh soft red fruits and 57 samples of other types of fresh produce (tomatoes, cucumber and fruit salads) were analyzed. Firstly, the NoV detection methodology, including virus and RNA extraction, real-time RT-PCR and quality controls were compared among the three countries. In addition, confirmation and genotyping of the NoV strains was attempted for a subset of NoV positive samples using conventional RT-PCR targeting an alternative region followed by sequencing. Analysis of the process control showed that 653, 179 and 18 samples of the leafy greens, soft red fruits and other fresh produce types were valid for analysis based on the recovery of the process control. NoV was detected by real-time RT-PCR in 28.2% (N = 641), 33.3% (N = 6) and 50% (N = 6) of leafy greens tested in Canada, Belgium and France, respectively. Soft red fruits were found positive by real-time RT-PCR in 34.5% (N = 29) and 6.7% (N = 150) of the samples tested in Belgium and France, respectively. 55.5% (N = 18) of the other fresh produce types, analyzed in Belgium, were found NoV positive by real-time RT-PCR. Conventional RT-PCR resulted in an amplicon of the expected size in 19.5% (52/266) of the NoV positive samples where this assay was attempted. Subsequent sequencing was only successful in 34.6% (18/52) of the suspected amplicons obtained by conventional RT-PCR. From this study, using the described methodology, NoV genomes were frequently detected in fresh produce however sequence confirmation was not successful for the majority of the samples tested. Infection or outbreaks were rarely or not known to be related to the NoV positive samples. With the increase in sensitivity of the detection methodology, there is an increasing concern about the interpretation of positive NoV results by real-time amplification. Strategies to confirm the results by real-time RT-PCR should be developed in analogy with the detection of microbial pathogens in foods. Detection might indicate contact with NoV in the fresh produce chain. Consequently, a potential risk for infection cannot be excluded but the actual risk from RT-PCR NoV positive produce is still unknown. Studies should be designed determining the probability of infection related to the presence or levels of NoV genomic copies.     

Hierarchical Bayesian analysis of censored microbiological contamination data for use in risk assessment and mitigation

Microbiological contamination data often is censored because of the presence of non-detects or because measurement outcomes are known only to be smaller than, greater than, or between certain boundary values imposed by the laboratory procedures. Therefore, it is not straightforward to fit distributions that summarize contamination data for use in quantitative microbiological risk assessment, especially when variability and uncertainty are to be characterized separately. In this paper, distributions are fit using Bayesian analysis, and results are compared to results obtained with a methodology based on maximum likelihood estimation and the non-parametric bootstrap method. The Bayesian model is also extended hierarchically to estimate the effects of the individual elements of a covariate such as, for example, on a national level, the food processing company where the analyzed food samples were processed, or, on an international level, the geographical origin of contamination data. Including this extra information allows a risk assessor to differentiate between several scenario’s and increase the specificity of the estimate of risk of illness, or compare different scenario’s to each other. Furthermore, inference is made on the predictive importance of several different covariates while taking into account uncertainty, allowing to indicate which covariates are influential factors determining contamination.     

Microbiological performance of a food safety management system in a food service operation

The microbiological performance of a food safety management system in a food service operation was measured using a microbiological assessment scheme as a vertical sampling plan throughout the production process, from raw materials to final product. The assessment scheme can give insight into the microbiological contamination and the variability of a production process and pinpoint bottlenecks in the food safety management system. Three production processes were evaluated: a high-risk sandwich production process (involving raw meat preparation), a medium-risk hot meal production process (starting from undercooked raw materials), and a low-risk hot meal production process (reheating in a bag). Microbial quality parameters, hygiene indicators, and relevant pathogens (Listeria monocytogenes, Salmonella, Bacillus cereus, and Escherichia coli O157) were in accordance with legal criteria and/or microbiological guidelines, suggesting that the food safety management system was effective. High levels of total aerobic bacteria (>3.9 log CFU/50 cm(2)) were noted occasionally on gloves of food handlers and on food contact surfaces, especially in high contamination areas (e.g., during handling of raw material, preparation room). Core control activities such as hand hygiene of personnel and cleaning and disinfection (especially in highly contaminated areas) were considered points of attention. The present sampling plan was used to produce an overall microbiological profile (snapshot) to validate the food safety management system in place.     

The reduction of murine norovirus 1, B. fragilis HSP40 infecting phage B40-8 and E. coli after a mild thermal pasteurization process of raspberry puree

Pasteurization processes of raspberry puree are nowadays limited to short times and rather low temperatures to maintain flavor and nutritional quality. Norovirus (NoV) outbreaks associated with raspberries highlight the need to determine the survival of NoV on this type of soft fruit. Therefore, resistance of murine norovirus 1 (MNV-1), a surrogate for human NoV, B. fragilis HSP40 infecting phage B40-8, and E. coli towards mild pasteurization was tested. Raspberry puree heat treated at 65 degrees C for 30s showed a 1.86, 2.77, and 3.89 log reduction of, respectively, MNV-1, E. coli, and B40-8. Heating at 75 degrees C for 15s established a 2.81 log reduction of MNV-1 while a 3.44 and 3.61 log reduction of B40-8 and E. coli was observed. No supplementary lethal effect of holding the heat-treated raspberry puree at 4 degrees C overnight was noticed. B40-8 failed to be useful as a tool to monitor NoV inactivation during mild pasteurization processes. Moreover, <3 log reductions of MNV-1 were observed suggesting that upon high initial contamination load, infectious NoV particles may remain on mildly pasteurized raspberry puree.     

Evaluation of viral extraction methods on a broad range of Ready-To-Eat foods with conventional and real-time RT-PCR for Norovirus GII detection

Noroviruses (NoV) are a common cause of foodborne outbreaks. In spite of that, no standard viral detection method is available for food products. Therefore, three viral elution-concentration methods and one direct RNA isolation method were evaluated on a broad range of Ready-To-Eat (RTE) food products (mixed lettuce, fruit salad, raspberries and two RTE dishes) artificially seeded with a diluted stool sample contaminated with NoV genogroup II. These seeding experiments revealed two categories of RTE products, fruits and vegetables grouped together and RTE dishes (penne and tagliatelle salads) which are rich in proteins and fat formed another category. The RNA extracts were amplified and detected with two conventional RT-PCR systems (Booster and Semi-nested GII) and one real-time RT-PCR (Real-time GII) assay. A fast direct RNA isolation method detected 10(2) RT-PCRU on 10 g penne and tagliatelle salads with the conventional RT-PCR assays. However real-time RT-PCR was less sensitive for penne salad. A viral elution-concentration method, including a buffer solution for the elution step and one polyethylene glycol (PEG) precipitation step, was able to detect 10(2) RT-PCRU on 50 g frozen raspberries with conventional and real-time RT-PCR assays. Moreover the latter extraction method used no environmental hazardous chemical reagents and was easy to perform.     

Antibacterial activity of Lactobacillus plantarum UG1 isolated from dry sausage: characterization, production and bactericidal action of plantaricin UG1

Lactobacillus plantarum UG1 isolated from dry sausage produced an antimicrobial substance that inhibited other strains of the genera Lactobacillus and Lactococcus, and some foodborne pathogens including Listeria monocytogenes, Bacillus cereus, Clostridium perfringens and Clostridium sporogenes. This antibacterial substance was inactivated by proteolytic enzymes and showed a bactericidal mode of action. Consequently, it was characterized as a bacteriocin, and was designated plantaricin UG1. This bacteriocin was stable in the pH range 4.5 to 7.0, partially inactivated by amylolytic enzymes and relatively thermostable. It was not affected by organic or lipolytic enzymes. Production of plantaricin UG1 was pH-and temperature-dependant and maximum yields were obtained in MRS broth cultures maintained at initial pH 6.5, and incubated at 25 °C to 30 °C, in the exponential to the early stationary growth phase of the producer organism. Ultrafiltration studies indicated that plantaricin UG1 has a molecular weight between 3 and 10 KDa. Curing experiments with L. plantarum UG1 resulted in the appearance of variants that lost bacteriocin production ability but were still immune to the bacteriocin. Plantaricin UG1 production appeared to be chromosomal encoded. Sensitive and insensitive Gram-positive bacteria adsorbed plantaricin UG1 irrespective of their susceptibility to it. In contrast, Gram-negative bacteria did not adsorb plantaricin UG1. The bactericidal action of plantaricin UG1 did not depend on the physiological state of the indicator culture and did not cause cell lysis. The resistance of two indicator strains to plantaricin UG1 has been studied.     

Evolution Of Antenna Sidelobe Regulation

Geostationary satellite networks, by their nature, operate in an interference environment. Engineering for an interference limited environment requires that earth station antennas be designed to minimize interference. To increase the efficiency of utilization of the satellite orbit significantly, the FCC has revised the antenna performance standards for transmitting antennas, more particularly in the first 9.20 from the main axis in the plane of the geostationary satellite orbit. These new FCC requirements are discussed in some detail.     

Effect of acid resistance of Escherichia coli O157:H7 on efficacy of buffered lactic acid to decontaminate chilled beef tissue and effect of modified atmosphere packaging on survival of Escherichia coli O157:H7 on red meat.

The present study examined the effect of pH-independent acid resistance of Escherichia coli O157:H7 on efficacy of buffered lactic acid to decontaminate chilled beef tissue. A varied level of acid resistance was observed among the 14 strains tested. Eight strains were categorized as acid resistant, four strains as acid sensitive, and two strains demonstrated acid-inducible acid resistance. The survival of an acid-resistant (II/45/4) and acid-sensitive (IX/8/16) E. coli O157:H7 strain on chilled beef tissue treated with 1 and 2% buffered lactic acid, sterile water, or no treatment (control) was followed. A gradual reduction of E. coli O157:H7 was noticed during the 10 days of storage at 4 degrees C for each of the treatments. Decontamination with 1 and 2% buffered lactic acid did not appreciably affect the pathogen. Differences in the pH-independent acid resistance of the strains had no effect on the efficacy of decontamination. The effect of modified atmosphere packaging (MAP) on survival of E. coli O157:H7 in red meat was also studied. MAP (40% CO2/60% N2) or vacuum did not significantly influence survival of E. coli O157:H7 on inoculated sliced beef (retail cuts) meat compared to packing in air. The relative small outgrowth of lactic acid bacteria during storage under vacuum for 28 days did not affect survival of E. coli O157:H7. Neither lactic acid decontamination nor vacuum or MAP packaging could enhance reduction of E. coli O157:H7 on beef, thus underlining the need for preventive measures to control the public health risk of E. coli O157:H7.