Implementation of hazard analysis and critical control point (HACCP) in yogurt production.

This study aimed to review hazard analysis and critical control points (HACCP) in the dairy industry for the production of yogurt. The food safety management system (FSMS) was implemented over the last several decades with several amendments. The need for practical and proactive procedures in the dairy industry was identified so that HACCP implementation could ensure that consumers would always have safe food. The concept of HACCP is a systemic and science-based method that can result in safe dairy products such as yogurt based on the complete analysis of manufacturing processes, recognition of hazards potentially present at all stages of production, and risk prevention. In yogurt production, raw milk receipt, pasteurization, packaging, and storage are the steps most susceptible to contamination and were considered critical control points. Further steps also need to be implemented to achieve other related control measures, and these will be discussed.

An electrochemical aptasensor for zearalenone detection based on the Co3O4/MoS2/Au nanocomposites and hybrid chain reaction.

An electrochemical aptasensor was used for the fast and sensitive detection of zearalenone (ZEN) based on the combination of Co3O4/MoS2/Au nanocomposites and the hybrid chain reaction (HCR). The glassy carbon electrode was coated with Co3O4/MoS2/Au nanomaterials to immobilize the ZEN-cDNA that had been bound with ZEN-Apt by the principle of base complementary pairing. In the absence of ZEN, the HCR could not be triggered because the ZEN-cDNA could not be exposed. After ZEN was added to the surface of the electrode, a complex structure was produced on the modified electrode by the combination of ZEN and ZEN-Apt. Therefore, the ZEN-cDNA can raise the HCR to produce the long-strand dsDNA structure. Due to the formation of dsDNA, the methylene blue (MB) could be inserted into the superstructure of branched DNA and the peak currents of the MB redox signal dramatically increased. So the concentration of ZEN could be detected by the change of signal intensity. Under optimized conditions, the developed electrochemical biosensing strategy showed an outstanding linear detection range of 1.0×10-10 mol/L to 1.0×10-6 mol/L, a low detection limit (LOD) of 8.5×10-11 mol/L with desirable selectivity and stability. Therefore, the fabricated platform possessed a great application potential in fields of food safety, medical detection, and drug analysis.

Case analysis of kinetics investigations in toxicity studies of pesticides to identify the nonlinearity of internal exposure and the influences of nonlinearity on the toxicological interpretation of pesticide residue.

The nonlinearity of internal exposure to 8 pesticides was investigated in toxicity studies using kinetics to identify nonlinearity visually and to investigate the influence of nonlinearity on toxicological evaluation. Data were obtained from risk assessment reports published by the Food Safety Commission (FSCJ). Nonlinearity was defined using 2 indicators: the lowest visual inflection point (LVIP) and the second lowest visual inflection point (SVIP) of kinetics by drawing a linear distribution chart. The area under the curve and 24-h urine concentrations were stable parameters used to identify the LVIP/SVIP. The sampling timing affected the blood concentrations, and the LVIP/SVIP was detected for 6 pesticides using the parent compounds or their metabolites as analytes. The subproportional nonlinearity was significant for these pesticides. The LVIP/SVIP values were consistent in the same species up to a 1-year period, but the values showed species-specific differences in several compounds. In all compounds found to be nonlinear, apical outcomes were observed at the SVIP or above. The presence of nonlinearity was recognized by the FSCJ. The recognition influenced their judgment of no-observed-adverse-effect levels (NOAELs) for carcinogenicity or health-based guidance values, indicating the importance of appropriate kinetics to identify the nonlinearity for toxicological evaluation of pesticide residue.

High-Efficiency Machine Learning Method for Identifying Foodborne Disease Outbreaks and Confounding Factors.

The China National Center for Food Safety Risk Assessment (CFSA) uses the Foodborne Disease Monitoring and Reporting System (FDMRS) to monitor outbreaks of foodborne diseases across the country. However, there are problems of underreporting or erroneous reporting in FDMRS, which significantly increase the cost of related epidemic investigations. To solve this problem, we designed a model to identify suspected outbreaks from the data generated by the FDMRS of CFSA. In this study, machine learning models were used to fit the data. The recall rate and F1-score were used as evaluation metrics to compare the classification performance of each model. Feature importance and pathogenic factors were identified and analyzed using tree-based and gradient boosting models. Three real foodborne disease outbreaks were then used to evaluate the best performing model. Furthermore, the SHapley Additive exPlanation value was used to identify the effect of features. Among all machine learning classification models, the eXtreme Gradient Boosting (XGBoost) model achieved the best performance, with the highest recall rate and F1-score of 0.9699 and 0.9582, respectively. In terms of model validation, the model provides a correct judgment of real outbreaks. In the feature importance analysis with the XGBoost model, the health status of the other people with the same exposure has the highest weight, reaching 0.65. The machine learning model built in this study exhibits high accuracy in recognizing foodborne disease outbreaks, thus reducing the manual burden for medical staff. The model helped us identify the confounding factors of foodborne disease outbreaks. Attention should be paid not only to the health status of those with the same exposure but also to the similarity of the cases in time and space.

Application of Whole-Genome Sequencing in the National Molecular Tracing Network for Foodborne Disease Surveillance in China.

National Molecular Tracing Network for Foodborne Disease Surveillance (TraNet) was launched in 2013, which is the only real-time whole-genome sequencing (WGS)-based subtyping network in China for effective foodborne disease surveillance. TraNet covers three levels of public health laboratories, national, provincial, and municipal. The TraNet national databases have a total of more than 54,000 entries representing seven common foodborne bacteria from humans, food, and environments. Raw sequence data are uploaded to TraNet by Data Delivery Center. Assembled sequence data, pulsed-field gel electrophoresis (PFGE) profiles, antibiotic resistance patterns, and epidemiological data are submitted to national pathogen-specific databases managed by China National Center for Food Safety Risk Assessment. PFGE patterns and WGS-based subtyping are compared for rapid differentiation of clusters of geographically diverse foodborne infections. WGS-based TraNet has played significant roles in improving foodborne disease surveillance in China for rapid outbreak investigation, source tracking, and cluster analysis of particular pathogens across the country.

A microbial challenge study for validating continuous radio-frequency assisted thermal processing pasteurization of egg white powder.

Spray dried egg white powder (EWP) is traditionally processed by hot room treatment for a prolonged period of time (67 °C for 15 days) to enhance its functionality (foaming and gelling) and to improve microbial safety of EWP. Our prior research demonstrated that radio-frequency (RF) assisted thermal processing can considerably reduce the processing time, without compromising the functional properties of EWP. In this study, continuous RF processing was evaluated for pasteurization of EWP. EWP samples were inoculated with a 5-strain Salmonella cocktail or Enterococcus faecium NRRL B-2354 for the microbial challenge studies. To evaluate the inoculation method, stability and homogeneity tests were conducted for both Salmonella and E. faecium in EWP. Continuous RF heating of EWP was conducted in a 6-kW, 27.12 MHz pilot-scale parallel-plate RF heating system. RF-assisted thermal processing of EWP at 80 °C for 2 h provided >6.69 log reduction for Salmonella. E. faecium was found to be a suitable surrogate for Salmonella due to its higher resistance and similar inactivation kinetics during RF heating of EWP. The validated RF-assisted thermal process can be scaled up for use in the egg industry.

Inkjet Printed Nanopatterned Aptamer-Based Sensors for Improved Optical Detection of Foodborne Pathogens.

The increasing incidence of infectious outbreaks from contaminated food and water supply continues imposing a global burden for food safety, creating a market demand for on-site, disposable, easy-to-use, and cost-efficient devices. Despite of the rapid growth of biosensors field and the generation of breakthrough technologies, more than 80% of the platforms developed at lab-scale never will get to meet the market. This work aims to provide a cost-efficient, reliable, and repeatable approach for the detection of foodborne pathogens in real samples. For the first time an optimized inkjet printing platform is proposed taking advantage of a carefully controlled nanopatterning of novel carboxyl-functionalized aptameric ink on a nitrocellulose substrate for the highly efficient detection of E. coli O157:H7 (25 colony forming units (CFU) mL-1 in pure culture and 233 CFU mL-1 in ground beef) demonstrating the ability to control the variation within ±1 SD for at least 75% of the data collected even at very low concentrations. From the best of the knowledge this work reports the lowest limit of detection of the state of the art for paper-based optical detection of E. coli O157:H7, with enough evidence (p > 0.05) to prove its high specificity at genus, species, strain, and serotype level.

Are food exposures obtained through commercial market panels representative of the general population? Implications for outbreak investigations.

Current methods of control recruitment for case-control studies can be slow (a particular issue for outbreak investigations), resource-intensive and subject to a range of biases. Commercial market panels are a potential source of rapidly recruited controls. Our study evaluated food exposure data from these panel controls, compared with an established reference dataset. Market panel data were collected from two companies using retrospective internet-based surveys; these were compared with reference data from the National Diet and Nutrition Survey (NDNS). We used logistic regression to calculate adjusted odds ratios to compare exposure to each of the 71 food items between the market panel and NDNS participants. We compared 2103 panel controls with 2696 reference participants. Adjusted for socio-demographic factors, exposure to 90% of foods was statistically different between both panels and the reference data. However, these differences were likely to be of limited practical importance for 89% of Panel A foods and 79% of Panel B foods. Market panel food exposures were comparable with reference data for common food exposures but more likely to be different for uncommon exposures. This approach should be considered for outbreak investigation, in conjunction with other considerations such as population at risk, timeliness of response and study resources.

Oriented assembly of surface plasmon resonance biosensor through staphylococcal protein A for the chlorpyrifos detection.

In this study, we report a direct surface plasmon resonance (SPR) biosensor based on an oriented assembly of antibody for the rapid detection of chlorpyrifos residue in agricultural samples. In this covalent-orientated strategy, staphylococcal protein A (SPA) was first covalently bound to the surface for monitoring chlorpyrifos residue, with subsequent binding of the antibody in an orientated fashion via its fragment crystallizable (Fc) region. Consequently, the SPA-modified biosensor exhibited a satisfactory specificity and a low detection limit of 0.056 ng mL-1 for chlorpyrifos, with a linear detection range of 0.25-50.0 ng mL-1. Under optimal conditions, the sensor chip could be regenerated for at least 210 cycles. The results presented here indicate that the SPA-modified sensor chip can successfully improve the sensitivity and obviating the need of the modification of the antibody. The developed SPR biosensor method has the great potential for rapid, sensitive, and specific detection with broad applications in areas of environmental monitoring and food safety. Graphical abstract.

Evaluation of new approaches to the access of official monitoring results for live bivalves molluscs.

BACKGROUND: Live bivalve molluscs, echinoderms, tunicates and marine gastropod are referred in EU food laws, and require member states to implement official controls in classified production areas, with the monitoring and classification of those areas. If, due to contaminant tests results, a production area is closed, any product from there is prohibited to be commercialized. Mobile applications optical character recognition (OCR) functionalities could ease the access to contaminant levels and production area classifications. This study verifies what information is available in live bivalves' labels, describes an OCR algorithm for those labels and evaluates it. METHODS: 86 labels were selected from four sale points in Lisbon, and photographed using smartphones. Each label was evaluated by a human to determine what data was available (either required or not). An OCR algorithm was developed and applied on the collected labels and validated against the data extracted by the human analysis. RESULTS: The analysis shows that all the labels included the required information, and 63% of the labels included the identifier for the production zone. The label-reading algorithm performs with an accuracy of 79.85% for the individual values. CONCLUSION: High accuracy of the developed label-reading algorithm shows potential for providing instant automatic access to the date and production area, but is affected by the variability on the label structure. Although not required by food laws, the majority of the sampled labels included complementary information (classified production area) that will allow access to more precise information about the existing biotoxin tests and analysis results.

Toxicological safety evaluation of the human-identical milk oligosaccharide 6'-sialyllactose sodium salt.

Human milk oligosaccharides (HMOs) are abundant in breastmilk, but their presence in infant formula is negligible. Sialylated HMOs, such as 6'-sialyllactose, constitute a significant portion of the HMO fraction of human milk and are linked to important biological functions. To produce infant formula that is more comparable with human milk, biosynthesized sialyllactoses known as human-identical milk oligosaccharides (structurally identical counterparts to their respective naturally occurring HMOs in breastmilk) are proposed for use in infant formula and other functional foods for the general population. To support the safety of 6'-sialyllactose sodium salt (6'-SL), a 90-day oral (gavage) toxicity study and in vitro genotoxicity tests were conducted. The 90-day study is the first to be conducted with 6'-SL using neonatal rats (day 7 of age at the start of dosing), thus addressing safety of 6'-SL for consumption by the most sensitive age group (infants). In the 90-day study, neonatal rats received 6'-SL at doses up to 5000 mg/kg body weight (BW)/day and reference controls received 5000 mg/kg BW/day of fructooligosaccharide (an ingredient approved for use in infant formula) for comparison with the high-dose 6'-SL group, followed by a 4-week recovery period. There was no evidence of genotoxicity in vitro. No test item-related adverse effects were observed on any parameter in the 90-day study, thus the high dose (5000 mg/kg BW/day) was established as the no-observed-adverse-effect level. These results confirm that 6'-SL is safe for use in formula milk for infants and in other functional foods for the general population.

Toxicological safety assessment of the human-identical milk oligosaccharide 3'-sialyllactose sodium salt.

Human breastmilk is a mixture of nutrients, hormones and bioactive molecules that are vital for infant growth and development. Infant formula (IF) lacks many of these compounds, most notably human milk oligosaccharides (HMOs), which are abundant in breastmilk but scarce in IF. Sialyllactoses, such as 3'-sialyllactose, constitute a large portion of the HMO fraction. To produce IF that matches breastmilk more closely, biosynthesized human-identical milk oligosaccharides (structurally identical to HMOs) such as 3'-sialyllactose sodium salt (3'-SL) are proposed for use in IF and foods for the general population. The safety assessment of 3'-SL comprised in vitro genotoxicity tests and a 90-day oral (gavage) toxicity study. This is the first 90-day study conducted with 3'-SL using neonatal rats (7 days old at the start of dosing-equivalent age to newborn human infants in terms of central nervous system and reproductive development), demonstrating the safety of 3'-SL for consumption by infants, the most sensitive age group. The neonatal rats received 3'-SL at doses up to 5,000 mg/kg body weight (BW)/day and reference controls received 5,000 mg/kg BW/day of fructooligosaccharide (an ingredient approved for use in IF) for comparison with the high-dose 3'-SL group, followed by a 4-week recovery period. There was no evidence of genotoxicity in vitro. In the absence of any test item-related adverse effects in the 90-day study, the high dose (5,000 mg/kg BW/day) was established as the no-observed-adverse-effect level. This confirms the safety of 3'-SL for use in IF for infants, as well as in functional foods for the general population.

Emerging Point-of-care Technologies for Food Safety Analysis.

Food safety issues have recently attracted public concern. The deleterious effects of compromised food safety on health have rendered food safety analysis an approach of paramount importance. While conventional techniques such as high-performance liquid chromatography and mass spectrometry have traditionally been utilized for the detection of food contaminants, they are relatively expensive, time-consuming and labor intensive, impeding their use for point-of-care (POC) applications. In addition, accessibility of these tests is limited in developing countries where food-related illnesses are prevalent. There is, therefore, an urgent need to develop simple and robust diagnostic POC devices. POC devices, including paper- and chip-based devices, are typically rapid, cost-effective and user-friendly, offering a tremendous potential for rapid food safety analysis at POC settings. Herein, we discuss the most recent advances in the development of emerging POC devices for food safety analysis. We first provide an overview of common food safety issues and the existing techniques for detecting food contaminants such as foodborne pathogens, chemicals, allergens, and toxins. The importance of rapid food safety analysis along with the beneficial use of miniaturized POC devices are subsequently reviewed. Finally, the existing challenges and future perspectives of developing the miniaturized POC devices for food safety monitoring are briefly discussed.

Optimizing the Use of Zebrafish Feeding Trials for the Safety Evaluation of Genetically Modified Crops.

In Europe, the toxicological safety of genetically modified (GM) crops is routinely evaluated using rodent feeding trials, originally designed for testing oral toxicity of chemical compounds. We aimed to develop and optimize methods for advancing the use of zebrafish feeding trials for the safety evaluation of GM crops, using maize as a case study. In a first step, we evaluated the effect of different maize substitution levels. Our results demonstrate the need for preliminary testing to assess potential feed component-related effects on the overall nutritional balance. Next, since a potential effect of a GM crop should ideally be interpreted relative to the natural response variation (i.e., the range of biological values that is considered normal for a particular endpoint) in order to assess the toxicological relevance, we established natural response variation datasets for various zebrafish endpoints. We applied equivalence testing to calculate threshold equivalence limits (ELs) based on the natural response variation as a method for quantifying the range within which a GM crop and its control are considered equivalent. Finally, our results illustrate that the use of commercial control diets (CCDs) and null segregant (NS) controls (helpful for assessing potential effects of the transformation process) would be valuable additions to GM safety assessment strategies.

Microbial bioinformatics for food safety and production.

In the production of fermented foods, microbes play an important role. Optimization of fermentation processes or starter culture production traditionally was a trial-and-error approach inspired by expert knowledge of the fermentation process. Current developments in high-throughput 'omics' technologies allow developing more rational approaches to improve fermentation processes both from the food functionality as well as from the food safety perspective. Here, the authors thematically review typical bioinformatics techniques and approaches to improve various aspects of the microbial production of fermented food products and food safety.

Carbon dots: Principles and their applications in food quality and safety detection.

In the past ten years, as a novel and prospective nanomaterials, carbon dots have acquired tremendous attention for their unique optical and physicochemical properties, high compatibility and low cost, as well as great potential in sensing area. This review aims to present the current detecting principles based on carbon dots and other nano biological technologies, involving fluorescence quenching and recovery mechanisms. The synthetic and modificatory approaches in making carbon dots including top-down and bottom-up methods, as well as surface passivation and heteroatom doping ways are introduced. Their applications in food area, concerning detection of nutrients, restricted or banned substances as well as foodborne pathogenic bacteria and the toxins secreted are discussed. Finally, the difficulties to be overcome or problems to be solved are presented, and other novel techniques to combine with carbon dots to obtain more stable and specific nanosensors in various fields are proposed. Although carbon dots based sensors have shown the potential in sensing aspect of food area, as food samples are complex in compositions that may cause interferences, more novel techniques are needed to combine with carbon dots to develop sensitive and specific sensing probes.

Elucidation of non-intentionally added substances migrating from polyester-polyurethane lacquers using automated LC-HRMS data processing.

An untargeted strategy aiming at identifying non-intentionally added substances (NIAS) migrating from coatings was developed. This innovative approach was applied to two polyester-polyurethane lacquers, for which suppliers previously provided the identity of the monomers involved. Lacquers were extracted with acetonitrile and analyzed by liquid chromatography-high resolution mass spectrometry (LC-HRMS). Data, acquired in the full scan mode, were processed using an open-source R-environment (xcms and CAMERA packages) to list the detected features and deconvolute them in groups related to individual compounds. The most intense groups, accounting for more than 85% of cumulated feature intensities, were then investigated. A homemade database, populated with predicted polyester oligomer combinations from a relevant selection of diols and diacids, enabled highlighting the presence of 14 and 17 cyclic predicted polyester oligomers in the two lacquers, including three mutual combinations explained by common known monomers. Combination hypotheses were strengthened by chromatographic considerations and by the investigation of fragmentation patterns. Regarding unpredicted migrating substances, four monomers were hypothesised to explain several polyester or caprolactam oligomer series. Finally, considering both predicted and tentatively elucidated unpredicted oligomers, it was possible to assign hypotheses to features representing up to 82% and 90% of the cumulated intensities in the two lacquers, plus 9% and 3% (respectively) originating from the procedural blank. Graphical abstract Elucidation of non-intentionally added substances.

Application and evaluation of a high-resolution mass spectrometry screening method for veterinary drug residues in incurred fish and imported aquaculture samples.

The ability to detect chemical contaminants, including veterinary drug residues in animal products such as fish, is an important example of food safety analysis. In this paper, a liquid chromatography high-resolution mass spectrometry (LC-HRMS) screening method using a quadrupole-Orbitrap instrument was applied to the analysis of veterinary drug residues in incurred tissues from aquacultured channel catfish, rainbow trout, and Atlantic salmon and imported aquacultured products including European eel, yellow croaker, and tilapia. Compared to traditional MS methods, the use of HRMS with nontargeted data acquisition and exact mass measurement capability greatly increased the scope of compounds that could be monitored simultaneously. The fish samples were prepared for analysis using a simple efficient procedure that consisted of an acidic acetonitrile extraction followed by solid phase extraction cleanup. Two different HRMS acquisition programs were used to analyze the fish extracts. This method detected and identified veterinary drugs including quinolones, fluoroquinolones, avermectins, dyes, and aminopenicillins at residue levels in fish that had been dosed with those compounds. A metabolite of amoxicillin, amoxicillin diketone, was also found at high levels in catfish, trout, and salmon. The method was also used to characterize drug residues in imported fish. In addition to confirming findings of fluoroquinolone and sulfonamide residues that were found by traditional targeted MS methods, several new compounds including 2-amino mebendazole in eel and ofloxacin in croaker were detected and identified. Graphical Abstract Aquacultured samples are analyzed with a high-resolution mass spectrometry screening method to detect and identify unusual veterinary drug residues including ofloxacin in an imported fish.

Analysis of a variety of inorganic and organic additives in food products by ion-pairing liquid chromatography coupled to high-resolution mass spectrometry.

A reversed-phase ion-pairing chromatographic method was developed for the detection and quantification of inorganic and organic anionic food additives. A single-stage high-resolution mass spectrometer (orbitrap ion trap, Orbitrap) was used to detect the accurate masses of the unfragmented analyte ions. The developed ion-pairing chromatography method was based on a dibutylamine/hexafluoro-2-propanol buffer. Dibutylamine can be charged to serve as a chromatographic ion-pairing agent. This ensures sufficient retention of inorganic and organic anions. Yet, unlike quaternary amines, it can be de-charged in the electrospray to prevent the formation of neutral analyte ion-pairing agent adducts. This process is significantly facilitated by the added hexafluoro-2-propanol. This approach permits the sensitive detection and quantification of additives like nitrate and mono-, di-, and triphosphate as well as citric acid, a number of artificial sweeteners like cyclamate and aspartame, flavor enhancers like glutamate, and preservatives like sorbic acid. This is a major advantage, since the currently used analytical methods as utilized in food safety laboratories are only capable in monitoring a few compounds or a particular category of food additives. Graphical abstract Deptotonation of ion pair agent in the electrospray interface.

Non-targeted analysis of unexpected food contaminants using LC-HRMS.

A non-target analysis method for unexpected contaminants in food is described. Many current methods referred to as "non-target" are capable of detecting hundreds or even thousands of contaminants. However, they will typically still miss all other possible contaminants. Instead, a metabolomics approach might be used to obtain "true non-target" analysis. In the present work, such a method was optimized for improved detection capability at low concentrations. The method was evaluated using 19 chemically diverse model compounds spiked into milk samples to mimic unknown contamination. Other milk samples were used as reference samples. All samples were analyzed with UHPLC-TOF-MS (ultra-high-performance liquid chromatography time-of-flight mass spectrometry), using reversed-phase chromatography and electrospray ionization in positive mode. Data evaluation was performed by the software TracMass 2. No target lists of specific compounds were used to search for the contaminants. Instead, the software was used to sort out all features only occurring in the spiked sample data, i.e., the workflow resembled a metabolomics approach. Procedures for chemical identification of peaks were outside the scope of the study. Method, study design, and settings in the software were optimized to minimize manual evaluation and faulty or irrelevant hits and to maximize hit rate of the spiked compounds. A practical detection limit was established at 25 µg/kg. At this concentration, most compounds (17 out of 19) were detected as intact precursor ions, as fragments or as adducts. Only 2 irrelevant hits, probably natural compounds, were obtained. Limitations and possible practical use of the approach are discussed.