Assessment of transcriptional reprogramming of lettuce roots in response to chitin soil amendment.

Chitin soil amendment is known to improve soil quality, plant growth and stress resilience, but the underlying mechanisms are not well understood. In this study, we monitored chitin's effect on lettuce physiology every two weeks through an eight-week growth period, analyzed the early transcriptional reprogramming and related metabolomic changes of lettuce, in response to crab chitin treatment in peat-based potting soil. In commercial growth conditions, chitin amendment still promoted lettuce growth, increased chlorophyll content, the number of leaves and crop head weight from week six. The flavonoid content in lettuce leaves was altered as well, showing an increase at week two but a decrease from week six. Transcriptomic analysis showed that over 300 genes in lettuce root were significantly differentially expressed after chitin soil treatment. Gene Ontology-term (GO) enrichment analysis revealed statistical overrepresentation of GO terms linked to photosynthesis, pigment metabolic process and phenylpropanoid metabolic process. Further analysis of the differentially expressed genes (DEGs) showed that the flavonoid pathway was mostly upregulated whereas the bifurcation of upstream phenylpropanoid pathway towards lignin biosynthesis was mostly downregulated. Metabolomic analysis revealed the upregulation of salicylic acid, chlorogenic acid, ferulic acid, and p-coumaric acid in chitin-treated lettuce seedlings. These phenolic compounds (PCs) mainly influence the phenylpropanoid biosynthesis pathway and may play important roles in plant defense reactions. Our results suggest that chitin soil amendments might activate induced resistance by priming lettuce plants and promote lettuce growth via transcriptional changes.

Quantitative contamination assessment of Escherichia coli in baby spinach primary production in Spain: Effects of weather conditions and agricultural practices.

A quantitative microbial contamination model of Escherichia coli during primary production of baby spinach was developed. The model included only systematic contamination routes (e.g. soil and irrigation water) and it was used to evaluate the potential impact of weather conditions, agricultural practices as well as bacterial fitness in soil on the E. coli levels present in the crop at harvest. The model can be used to estimate E. coli contamination of baby spinach via irrigation water, via soil splashing due to irrigation water or rain events, and also including the inactivation of E. coli on plants due to solar radiation during a variable time of culturing before harvest. Seasonality, solar radiation and rainfall were predicted to have an important impact on the E. coli contamination. Winter conditions increased E. coli prevalence and levels when compared to spring conditions. As regards agricultural practices, both water quality and irrigation system slightly influenced E. coli levels on baby spinach. The good microbiological quality of the irrigation water (average E. coli counts in positive water samples below 1 log/100mL) could have influenced the differences observed among the tested agricultural practices (water treatment and irrigation system). This quantitative microbial contamination model represents a preliminary framework that assesses the potential impact of different factors and intervention strategies affecting E. coli concentrations at field level. Taking into account that E. coli strains may serve as a surrogate organism for enteric bacterial pathogens, obtained results on E. coli levels on baby spinach may be indicative of the potential behaviour of these pathogens under defined conditions.

Analysis of domestic refrigerator temperatures and home storage time distributions for shelf-life studies and food safety risk assessment.

In the framework of food safety, when mimicking the consumer phase, the storage time and temperature used are mainly considered as single point estimates instead of probability distributions. This singlepoint approach does not take into account the variability within a population and could lead to an overestimation of the parameters. Therefore, the aim of this study was to analyse data on domestic refrigerator temperatures and storage times of chilled food in European countries in order to draw general rules which could be used either in shelf-life testing or risk assessment. In relation to domestic refrigerator temperatures, 15 studies provided pertinent data. Twelve studies presented normal distributions, according to the authors or from the data fitted into distributions. Analysis of temperature distributions revealed that the countries were separated into two groups: northern European countries and southern European countries. The overall variability of European domestic refrigerators is described by a normal distribution: N (7.0, 2.7)°C for southern countries, and, N (6.1, 2.8)°C for the northern countries. Concerning storage times, seven papers were pertinent. Analysis indicated that the storage time was likely to end in the first days or weeks (depending on the product use-by-date) after purchase. Data fitting showed the exponential distribution was the most appropriate distribution to describe the time that food spent at consumer's place. The storage time was described by an exponential distribution corresponding to the use-by date period divided by 4. In conclusion, knowing that collecting data is time and money consuming, in the absence of data, and at least for the European market and for refrigerated products, building a domestic refrigerator temperature distribution using a Normal law and a time-to-consumption distribution using an Exponential law would be appropriate.

Measuring general animal health status: Development of an animal health barometer.

The development of an animal health barometer, an instrument to measure the general health of the Belgian livestock population on a yearly basis and to monitor its evolution over time, is described. The elaboration of a set of 13 animal health indicators (AHIs) as the basis for the animal health barometer is discussed. These indicators were weighted by experts - including scientists, policy makers and agro-industrial representatives - to determine their relative weight in the barometer. The result of the barometer is expressed as a comparison with a previous year. Based on the results of the 13 AHIs, it is concluded that general animal health in Belgium shows a positive evolution since 2008. The animal health barometer provides a composite view of the status of livestock health in Belgium and is a tool to communicate in an intelligible, comprehensible manner on aspects of animal health to consumers and professional stakeholders in the animal production and food chain. Together with the food safety barometer (Baert et al., 2011. Food Res. Int. 44, 940) and the plant health barometer (Wilmart et al., 2014. Eur. J. Plant Pathol. doi: 10.1007/s10658-014-0547-x), the animal health barometer is one of the three instruments to provide a holistic view on the overall status of the safety of the food chain in Belgium.

Detection of noroviruses in shellfish and semiprocessed fishery products from a Belgian seafood company.

Shellfish have been implicated in norovirus (NoV) infection outbreaks worldwide. This study presents data obtained from various batches of shellfish and fishery products from a Belgian seafood company over a 6-month period. For the intact shellfish (oysters, mussels, and clams), 21 of 65 samples from 12 of 34 batches were positive for NoVs; 9 samples contained quantitative NoV levels at 3,300 to 14,300 genomic copies per g. For the semiprocessed fishery products (scallops and common sole rolls with scallop fragments), 29 of 36 samples from all eight batches were positive for NoVs; 17 samples contained quantitative NoV levels at 200 to 1,800 copies per g. This convenience study demonstrated the performance and robustness of the reverse transcription quantitative PCR detection and interpretation method and the added value of NoV testing in the framework of periodic control of seafood products bought internationally and distributed by a Belgian seafood processing company to Belgian food markets.

Microbiological quality and safety assessment of lettuce production in Brazil.

The microbiological quality and safety of lettuce during primary production in Brazil were determined by enumeration of hygiene indicators Escherichia coli, coliforms and enterococci and detection of enteric pathogens Salmonella and E. coli O157:H7 in organic fertilizers, soil, irrigation water, lettuce crops, harvest boxes and worker's hands taken from six different lettuce farms throughout the crop growth cycle. Generic E. coli was a suitable indicator for the presence of Salmonella and E. coli O157:H7, while coliforms and enterococci were not. Few pathogens were detected: 5 salmonellae and 2 E. coli O157:H7 from 260 samples, of which only one was lettuce and the others were manure, soil and water. Most (5/7) pathogens were isolated from the same farm and all were from organic production. Statistical analysis revealed the following environmental and agro-technical risk factors for increased microbial load and pathogen prevalence in lettuce production: high temperature, flooding of lettuce fields, application of contaminated organic fertilizer, irrigation with water of inferior quality and large distances between the field and toilets. Control of the composting process of organic fertilizers and the irrigation water quality appear most crucial to improve and/or maintain the microbiological quality and safety during the primary production of lettuce.

Microbiological quality and safety assessment of the Rwandan milk and dairy chain.

Milk is a valuable and nutritious food product that can partially fulfill the rising food demand of the growing African population. The microbiological status of milk and derived products was assessed throughout the milk and dairy chain in Rwanda by enumeration of the total mesophilic count, coliforms, and Staphylococcus aureus and detection of Salmonella and Listeria monocytogenes. The quality of raw milk was satisfactory for the majority of samples, but 5.2% contained Salmonella. At the processing level, the total mesophilic count and coliform numbers indicated ineffective heat treatment during pasteurization or postpasteurization contamination. Increasing bacterial counts were observed along the retail chain and could be attributed to insufficient temperature control during storage. Milk and dairy products sold in milk shops were of poor and variable microbiological quality in comparison with the pasteurized milk sold in supermarkets. In particular, the microbiological load and pathogen prevalence in cheese were unacceptably high.

A quantitative microbiological exposure assessment model for Bacillus cereus in REPFEDs.

One of the pathogens of concern in refrigerated and processed foods of extended durability (REPFED) is psychrotrophic Bacillus cereus, because of its ability to survive pasteurisation and grow at low temperatures. In this study a quantitative microbiological exposure assessment (QMEA) of psychrotrophic B. cereus in REPFEDs is presented. The goal is to quantify (i) the prevalence and concentration of B. cereus during production and shelf life, (ii) the number of packages with potential emetic toxin formation and (iii) the impact of different processing steps and consumer behaviour on the exposure to B. cereus from REPFEDs. The QMEA comprises the entire production and distribution process, from raw materials over pasteurisation and up to the moment it is consumed or discarded. To model this process the modular process risk model (MPRM) was used (Nauta, 2002). The product life was divided into nine modules, each module corresponding to a basic process: (1) raw material contamination, (2) cross contamination during handling, (3) inactivation during preparation, (4) growth during intermediate storage, (5) partitioning of batches in portions, (6) mixing portions to create the product, (7) recontamination during assembly and packaging, (8) inactivation during pasteurisation and (9) growth during shelf life. Each of the modules was modelled and built using a combination of newly gathered and literature data, predictive models and expert opinions. Units (batch/portion/package) with a B. cereus concentration of 10(5)CFU/g or more were considered 'risky' units. Results show that the main drivers of variability and uncertainty are consumer behaviour, strain variability and modelling error. The prevalence of B. cereus in the final products is estimated at 48.6% (±0.01%) and the number of packs with too high B. cereus counts at the moment of consumption is estimated at 4750 packs per million (0.48%). Cold storage at retail and consumer level is vital in limiting the exposure. Four key points were identified (i) raw material contamination, (ii) recontamination during packaging, (iii) reduction during pasteurisation and cooking and (iv) cold storage at retail and consumer level.

Assessment of the microbial safety and quality of cooked chilled foods and their production process.

Refrigerated processed foods of extended durability (REPFEDs) are a heterogeneous group of food products. This study assesses the microbial safety and quality along the production process in five REPFED companies. Samples were taken of raw materials (n=123), intermediate products (n=123), end products at production day (n=45) and at end of shelf life (n=90), food contact surfaces (n=226) and worker's hands/gloves (n=92). Samples are analysed for total psychrotrophic aerobic count, aerobic spore count, sulphite reducing Clostridia, Bacillus cereus and Listeria monocytogenes. Both L. monocytogenes and B. cereus were detected on the raw materials. Nine of 72 raw materials tested were positive (in 25g) for L. monocytogenes and all but one of these raw materials were raw or minimally processed animal products. Three of 123 raw materials contained high counts (>4log CFU/g) of B. cereus, all of these samples were dried herbs. During production both food contact surfaces (90/226) and gloves (43/92) contained increased levels of total psychrotrophic aerobic counts (≥3log CFU/25cm(2)). This points out a potential source of bacterial recontamination. However, only a four and six of 223 food contact surfaces were positive (per 25cm(2)) for L. monocytogenes and B. cereus respectively. None of the gloves sampled contained L. monocytogenes and only 2 sets of gloves were positive for B. cereus. Of the 123 intermediate products tested twelve tested positive for L. monocytogenes (in 25g) and 5 showed elevated counts of B. cereus (ca. 2.5log CFU/g). Despite the presence of L. monocytogenes in the raw materials, the production area and in some of the intermediate products, none of the end products were positive for L. monocytogenes and only 9 of 135 samples (6.7%) showed to have low numbers of B. cereus (<2.7log CFU/g). This results show that the current pasteurization processes and the food safety management system are adequate to guarantee the production of microbiologically safe foods but that some improvements can still be made with regard to supplier selection, cleaning and disinfection, hygiene training and setting the shelf life duration.

Insight into the prevalence and distribution of microbial contamination to evaluate water management in the fresh produce processing industry.

This study provided insight into the degree of microbial contamination in the processing chain of prepacked (bagged) lettuce in two Belgian fresh-cut produce processing companies. The pathogens Salmonella and Listeria monocytogenes were not detected. Total psychrotrophic aerobic bacterial counts (TPACs) in water samples, fresh produce, and environmental samples suggested that the TPAC is not a good indicator of overall quality and best manufacturing practices during production and processing. Because of the high TPACs in the harvested lettuce crops, the process water becomes quickly contaminated, and subsequent TPACs do not change much throughout the production process of a batch. The hygiene indicator Escherichia coli was used to assess the water management practices in these two companies in relation to food safety. Practices such as insufficient cleaning and disinfection of washing baths, irregular refilling of the produce wash baths with water of good microbial quality, and the use of high product/water ratios resulted in a rapid increase in E. coli in the processing water, with potential transfer to the end product (fresh-cut lettuce). The washing step in the production of fresh-cut lettuce was identified as a potential pathway for dispersion of microorganisms and introduction of E. coli to the end product via cross-contamination. An intervention step to reduce microbial contamination is needed, particularly when no sanitizers are used as is the case in some European Union countries. Thus, from a food safety point of view proper water management (and its validation) is a critical point in the fresh-cut produce processing industry.

Sensitivity analysis of a two-dimensional quantitative microbiological risk assessment: keeping variability and uncertainty separated.

The aim of quantitative microbiological risk assessment is to estimate the risk of illness caused by the presence of a pathogen in a food type, and to study the impact of interventions. Because of inherent variability and uncertainty, risk assessments are generally conducted stochastically, and if possible it is advised to characterize variability separately from uncertainty. Sensitivity analysis allows to indicate to which of the input variables the outcome of a quantitative microbiological risk assessment is most sensitive. Although a number of methods exist to apply sensitivity analysis to a risk assessment with probabilistic input variables (such as contamination, storage temperature, storage duration, etc.), it is challenging to perform sensitivity analysis in the case where a risk assessment includes a separate characterization of variability and uncertainty of input variables. A procedure is proposed that focuses on the relation between risk estimates obtained by Monte Carlo simulation and the location of pseudo-randomly sampled input variables within the uncertainty and variability distributions. Within this procedure, two methods are used-that is, an ANOVA-like model and Sobol sensitivity indices-to obtain and compare the impact of variability and of uncertainty of all input variables, and of model uncertainty and scenario uncertainty. As a case study, this methodology is applied to a risk assessment to estimate the risk of contracting listeriosis due to consumption of deli meats.

Modeling logistic performance in quantitative microbial risk assessment.

In quantitative microbial risk assessment (QMRA), food safety in the food chain is modeled and simulated. In general, prevalences, concentrations, and numbers of microorganisms in media are investigated in the different steps from farm to fork. The underlying rates and conditions (such as storage times, temperatures, gas conditions, and their distributions) are determined. However, the logistic chain with its queues (storages, shelves) and mechanisms for ordering products is usually not taken into account. As a consequence, storage times-mutually dependent in successive steps in the chain-cannot be described adequately. This may have a great impact on the tails of risk distributions. Because food safety risks are generally very small, it is crucial to model the tails of (underlying) distributions as accurately as possible. Logistic performance can be modeled by describing the underlying planning and scheduling mechanisms in discrete-event modeling. This is common practice in operations research, specifically in supply chain management. In this article, we present the application of discrete-event modeling in the context of a QMRA for Listeria monocytogenes in fresh-cut iceberg lettuce. We show the potential value of discrete-event modeling in QMRA by calculating logistic interventions (modifications in the logistic chain) and determining their significance with respect to food safety.