Food Safety Research Research Articles

Genomic and phenotypic polymorphism of Clostridium botulinum Group II strain Beluga through laboratory domestication

Laboratory domestication is the result of genetic and physiological changes of organisms acquired during numerous passages in vitro. This phenomenon has been observed in bacteria as well as in higher organisms. In an effort to understand the impact of laboratory domestication on the foodborne pathogen Clostridium botulinum and related microbial food safety research, we investigated multiple spore stocks of C. botulinum Group II Beluga from our collection, as that is a widely applied model strain used in laboratories over decades. An acquired nutrient auxotrophy was confirmed as thymidine dependency using phenotypic microarrays. In parallel, whole-genome re-sequencing of all stocks revealed a mutation in thyA encoding thymidylate synthase essential for de-novo synthesis of dTMP from dUMP in the auxotrophic stocks. A thyA-deficient Beluga variant stock was successfully complemented by introducing an intact variant of thyA and thymidine prototrophy was restored, indicating that the thymidine auxotrophy was solely due to the presence of a SNP in thyA. Our data suggested that this mutation, deleterious under nutrient-poor growth conditions in a chemically defined medium, has been present and maintained in laboratory stocks for nearly 30 years. Yet, the mutation remained unidentified since receiving the strain, most likely due to routine use of culture conditions optimized for growth performance. This work pinpoints the need for careful monitoring of model strains extensively used in laboratory settings at both phenotypic and genomic level. In applications like food safety challenge tests, compromised strains could cause incorrect predictions and thereby have deleterious consequences. To mitigate the risk of acquiring mutations, we recommend keeping passage numbers of laboratory strains low and to avoid single-colony passaging. In addition, relevant strains should be subjected to regular WGS checks and physiological validation to exclude DNA mutations with potential negative impacts on research data integrity and reproducibility.

Modernization Data Analysis and Visualization for Food Safety Research Outcomes

Appropriate data collection and using reliable and accurate procedures are the first steps in conducting an experiment that will provide trustworthy outcomes. It is key to perform an assertive statistical analysis and data visualization for a correct interpretation and communication of results. A clear statistical summary and presentation of the data is critical for the reader to easily process and comprehend experimental results. Nowadays, there are a series of different tools to perform proper statistical analysis and create elaborate graphs that will help readers to understand the data, identify trends, detect outliers, evaluate statistical outputs, etc. However, researchers that are beginning to navigate experiments do not frequently encounter a guide that can provide basic principal concepts to begin their statistical analysis and data presentation. Therefore, the objective of this article is to provide a guide or manual to analyze and presents results focused on different types of common food safety experiments, including method comparisons, intervention studies, pathogen presence experiments, bio-mapping, statistical process control, and shelf life experiments. This review will provide information about data visualization options and statistical analysis approaches for different food safety experiments. In addition, basic concepts about descriptive statistics and possible solutions for issues related to microbiological measurements will be discussed.

Development and evaluation of a modified most probable number (MPN) method for enumerating rifampicin‐resistant Escherichia coli in agricultural, food, and environmental samples

AbstractLow level of rifampicin‐resistant E. coli is frequently used in food safety research as a model microorganism and is typically enumerated using traditional Most Probable Number (MPN) method. To simplify the process, we developed and validated a modified MPN method based on lactose peptone broth (LPB) containing rifampicin and bromocresol purple, the broth changes to a yellow color due to pH shifts induced by lactose fermentation from E. coli. E. coli TVS353 were prepared in following suspensions including healthy cells, injured cells, competitive bacteria with or without rifampicin resistance, and unknown natural microflora. E. coli TVS353 was quantified by traditional MPN method and our improved method. At a predicted level of 1 log CFU/mL, there was no significant difference in the MPN index of LPBR compared with TSB and TSBR in these suspensions (p > 0.05). Confirmation results generated from this new color change MPN method were analogous to traditional MPN methods.

Navigating Leadership and Communication: A Study of Emerging Leaders in Indian Organizations

Introduction: The research explores the dynamic interplay between the leadership styles and communication strategies in Indian context. In order to understand the connection between leadership and communication this research will use SPSS. 
 Literature review: This section delves into unraveling the diverse spectrum of leadership styles that are prevalent in Indian organizations. The importance of the problem-solving and informational-sharing studies has emphasized the complexity of the evaluative steps that have guided the theoretical framework to enable the research methodology.
 Methodology: The primary quantitative methodology employs a survey tool which has been administered on the 55 participants. SPSS solution has recognised so many attributes that have benefitted the organizational outcomes. The application of correlation, descriptive analysis and ANOVA has accessorized the vital values by making qualitative and quantitative assessments yielding a broad perception.
 Findings: This section illuminates the prevalence of diverse leadership styles incorporating transformational leaderships. It adopts a clear and delicate model which places emphasis on women as key players to the economic emancipation of citizens. From the findings, it has been obtained that there is a significant relationship between employee commitment and organizational success which is -0.74 at 0.01 level of significance.
 Discussion: From the discussion section, it has been examined that for organizational practices and leadership developments, as a result of these actionable insights, the young leader in India has been able to initiate the social media revolution. Moreover, tailoring a clear and transparent organization has emerged as a common factor to acknowledge the quantitative approaches
 Conclusion: The conclusion restates the main findings, which would in turn conclude any contributions to food safety research which should be derived from the study. Moreover, the dynamics and organizations landscape has been considered to be increasingly successful in empowering the leaders by means of expertise irrespective of relevance. Such strategy works through the complex nature of Indian corporate and organizational paradigm principles, which provides the foundation for leadership development programs to be conducted in future among researchers broadly.
 

Mircrofabricating double-sided polydimethylsiloxane (PDMS) artificial phylloplane for microbial food safety research

Leafy green surface microbiology studies often experience significant variations in results due to the heterogeneous nature of leaf surfaces. To provide a precise and controllable substitute, we microfabricated double-sided artificial leafy green phylloplanes using polydimethylsiloxane (PDMS) with a vinyl-terminated polyethylene glycol chain-based hydrophobicity modifier (PDMS-PEG) to modify PDMS hydrophobicity. We further tested the properties and applications of these artificial leaves, by examining the function of epicuticular wax, growth and survival of E. coli O157:H7 87–23 on the surface, and removal of attached E. coli cells via sanitation. The double-sided PDMS-PDMS-PEG leaves well-replicated their natural counterparts in macroscopic and microscopic structure, hydrophobicity, and E. coli O157:H7 87–23 attachment. After depositing natural epicuticular wax onto artificial leaves, the leaf surface wetting ability decreased, while E. coli O157:H7 87–23 surface retention increased. The artificial leaves supplied with lettuce lysate or bacterial growth media supported E. coli O157:H7 87–23 growth and survival similarly to those on natural leaves. In the sanitation test, the artificial lettuce leaves also displayed patterns similar to those of natural leaves regarding sanitizer efficiency. Overall, this study showcased the microfabrication and applications of double-sided PDMS-PDMS-PEG leaves as a replicable and controllable platform for future leafy green food safety studies.

The risk of hazards in laboratories of educational groups in the Faculty of Health, Shahid Sadougi University of Medical Sciences, Yazd-2023

Introduction: Improving the safety of working environments is one of the duties of any efficient system. Evaluating the risk of hazards is one of the most efficient ways for increasing safety important issues. Paying attention to the statistics of laboratory accidents and the positive effects of risk assessment in order to improve safety is of great importance. Materials and Methods: The present study was practical in terms of purpose, and cross-sectional in terms of descriptive-analytical method, which to evaluate the laboratories of Shahid Sadoqhi University of Medical Sciences, Yazd, using a quantitative three-dimensional cost equation. Based on the regular observation of existing processes and equipment, the evaluation team used a quantitative risk equation as an indication of safety performance at the level of laboratories. The ranking was based on the probability of risk, the frequency of effective exposure, and the maximum damage potential through a quantitative equation. Finally, the risk rate score of each laboratory was determined and the ranking of the risk level of the laboratories was announced. Suggested corrective solutions were presented and the evaluation was repeated. After collecting the data, it was entered into SPSS 20 software and interpreted using descriptive and analytical statistics. Results: In general, 52% of the risks identified in laboratories are at a low risk level. The highest and lowest average evaluation scores were 55.86 ± 30.89 for environmental chemistry and 28.38 ± 13.51 for food safety and health research laboratory, respectively. The highest evaluation score was given to the environment laboratory. There was a significant relationship between the risk components before and after providing the suggested solutions. Conclusion: According to the results of the research, the overall safety status of the educational laboratories in the Faculty of Health is at a relatively favorable level. However, chemical hazards and thermal safety are the highest priority to control and perform better regarding improvement.

A high-throughput gut-on-chip platform to study the epithelial responses to enterotoxins

Enterotoxins are a type of toxins that primarily affect the intestines. Understanding their harmful effects is essential for food safety and medical research. Current methods lack high-throughput, robust, and translatable models capable of characterizing toxin-specific epithelial damage. Pressing concerns regarding enterotoxin contamination of foods and emerging interest in clinical applications of enterotoxins emphasize the need for new platforms. Here, we demonstrate how Caco-2 tubules can be used to study the effect of enterotoxins on the human intestinal epithelium, reflecting toxins’ distinct pathogenic mechanisms. After exposure of the model to toxins nigericin, ochratoxin A, patulin and melittin, we observed dose-dependent reductions in barrier permeability as measured by TEER, which were detected with higher sensitivity than previous studies using conventional models. Combination of LDH release assays and DRAQ7 staining allowed comprehensive evaluation of toxin cytotoxicity, which was only observed after exposure to melittin and ochratoxin A. Furthermore, the study of actin cytoskeleton allowed to assess toxin-induced changes in cell morphology, which were only caused by nigericin. Altogether, our study highlights the potential of our Caco-2 tubular model in becoming a multi-parametric and high-throughput tool to bridge the gap between current enterotoxin research and translatable in vivo models of the human intestinal epithelium.

Draft genomic sequence for uropathogenic Staphylococcus saprophyticus ATCC 49453.

Staphylococcus saprophyticus, a common contaminant of foods, causes urinary tract infections in humans. Here, we report the draft genomic sequence for S. saprophyticus ATCC 49453, which is currently being used in food safety research.

Patulin removal in fruit-based products using probiotics and potential probiotics

<p>Patulin (PAT) is a prevalent mycotoxin frequently found in fruit and its derivatives, such as apple, pear, and juices. Despite worldwide attempts to diminish the levels of PAT at every stage of the fruit production process, its contamination rate remains high. This mycotoxin is worrisome due to its potential adverse impacts on human health. Eating PAT-contaminated fruit can lead to acute and chronic health issues. It is established, by the Joint FAO/WHO, a maximum tolerable daily intake for PAT at 0.4 µg/kg/day. Therefore, monitoring for PAT contamination is essential for the safe consumption of fruits and fruit-related products such as juices, purees, ciders, jams, marmalades, vinegar, and dried fruits. PAT has physiochemical properties that enable its survival in cold, hypoxic, acidic, or high-temperature conditions. Ideally, detoxification procedures should aim to reduce the level of toxins to safe levels whilst preserving the nutritional and palatable values of the treated commodity. There are several physical, chemical, and biological techniques available for PAT detoxification. However, while physical and chemical methods can remove PAT, they may also lower the nutrient quality and organoleptic properties of the food. Biological detoxification is an effective, environmentally friendly, easy, and cost-effective method, as established by existing research. It has proven efficacy in food safety research and regulatory compliance programs. Probiotics have been studied for their potential to reduce PAT in foods via different mechanisms (such as adsorption, degradation, and transformation), as well as all their health-beneficial effects. In this review, the reduction of PAT in fruit-based products using probiotics or potential probiotics is widely discussed.<strong></strong></p>

Methods and measures in food service food safety research: A review of the published literature

PurposeFood handlers working in food service establishments need to perform appropriate and consistent food safety practices to ensure that the prepared food is safe for consumers to eat, as a failure to do so may lead to foodborne illness. It is important to have an in-depth understanding of food handler food safety behaviour in the food service sector to design targeted educational interventions and approaches to safeguard consumers. The purpose of this review is to provide insight to the existing body of research regarding food handler food safety in the food service sector, and to identify future research needs in terms of methods and measures. MethodsThis study is a review of published food safety research studies (n = 118) focusing on food handlers in the food service sector conducted between 2001 and 2021. Data were collated regarding the methods used and the measures assessed in published studies to identify the gaps in current knowledge related to food safety in food service. FindingsThe majority of studies were conducted in the United States (29%), Brazil (17%) and United Kingdom (7%). A large proportion were carried out in restaurants (70%). Less than a third of studies (28%) relied on a mixed method approach for data collection; and only 12% of studies assessed a combination of cognitive and behavioural measures. Observation was used in only 29% of studies. A lack of observational data detailing food safety practices such as handwashing, temperature control, separation and cleaning was determined. SignificanceSuch findings indicate a greater need for mixed method approach in future food safety research. It is recommended that the understanding of food handler food safety performance is best obtained through a combined assessment of cognitive and behavioural measures with subsequent triangulation of findings to reduce bias and to obtain reliable results.

First report for fluorometric determination of kasugamycin based on amino acid-functionalized bimetallic nanoclusters

A rapid and straightforward turn-off fluorescence sensor has been developed for the detection of kasugamycin (KASN) using tryptophan-capped silver/gold bimetallic nanoclusters (Trp@Ag-Au NCs). The sensor's parameters, including reaction time and solvent type, were optimized to enhance its performance. The fluorescence emission of Trp@Ag-Au NCs is quenched upon the addition of KASN due to the aggregation of the Trp@Ag-Au NCs induced by the electrostatic and non-covalent interactions between the negatively charged Trp@Ag-Au NCs and the positively charged KASN. The sensor exhibited excellent sensitivity for KASN detection, demonstrating a linear concentration range from 0.5 to 500 µM and a low limit of detection (LOD) of 0.14 µM, determined at a signal-to-noise ratio of 3. These values indicate the sensor's ability to accurately detect KASN even at low concentrations. To evaluate its practical utility, spike recovery analysis was performed in synthetic samples, confirming its effectiveness in real-world sample matrices. The optimized sensor parameters, wide linear range, low LOD, and successful application in various samples highlight the sensor's potential for practical use in various applications, including food safety and biomedical research. Its simplicity, rapidity, and sensitivity make it a promising tool for the detection of KASN.

Boron nitride nanoplate-based strand exchange amplification with enhanced sensitivity and rapidity for quantitative detection of Staphylococcus aureus in food samples.

Staphylococcus aureus is one of the most common foodborne pathogens that can cause serious food poisoning and infectious diseases in humans. Standard identification approaches include nucleic acid amplification, but current amplification tools suffer from low amplification efficiency, resulting in the risk of low sensitivity and long detection time. Herein, boron nitride nanoplates (BNNPs) were chosen as an additive for enhancing the sensitivity and rapidity of strand exchange amplification (SEA), thereby successfully expanding the application of nucleic acid detection for detecting Staphylococcus aureus in food samples. As a result, SEA based on boron nitride nanoplates (BNNP-SEA) was employed for sensitive and rapid detection of foodborne pathogen Staphylococcus aureus. Compared with classical SEA, the BNNP-based SEA assay was more than 10-fold sensitive, and the detection time was reduced by 15 minutes. The optimized BNNP-based SEA shows a wide linear range from 40 pg to 50 ng in a diluted solution of the target DNA with a low detection limit of 40 pg. Moreover, the BNNP-based SEA achieves the quantitative detection of Staphylococcus aureus in different food samples (pork, beef, mutton, duck, milk and shrimp). In contrast to the classical SEA, the BNNP-based SEA method enabled sensitive and rapid detection of Staphylococcus aureus in the above food samples at concentrations as low as 5 × 103 CFU mL-1. The BNNP-based SEA assay is specific, sensitive and reliable, offering a valuable diagnostic technology for routine analysis in food safety research.

Holistic insights into carbon nanotubes and MXenes as a promising route to bio-sensing applications.

Essential biosensor use has become increasingly important in drug discovery and recognition, biomedicine, food safety, security, and environmental research. It directly contributed to the development of specialized, reliable diagnostic instruments known as biosensors, which use biological sensing components. Traditional biosensors have poor performance, so scientists need to develop advanced biosensors with promising selectivity, sensitivity, stability, and reusability. These are all parameter modifications associated with the characteristics of the sensing material. Carbon nanotubes (CNTs) and MXenes are promising as targeted sensing agents in advanced functional materials because of their promising chemical and physical properties and limited toxic effects. Based on available data and sensing performance, MXene is better for biosensing applications than CNTs. Because of their large specific surface area (SSA), superior electrical conductivity, and adaptable surface chemistry that facilitates simple functionalization and robust interactions with biomolecules, MXenes are typically regarded as the superior option for biosensors. Additionally, because of their hydrophilic nature, they are more suited to biological settings, which increases their sensitivity and efficacy in identifying biological targets. MXenes are more suitable for biosensing applications due to their versatility and compatibility with aquatic environments, even if CNTs have demonstrated stability and muscular mechanical strength. However, MXenes offer better thermal stability, which is crucial for applications in diverse temperature environments. This study reviews and compares the biosensing capabilities, synthesis methods, unique properties, and toxicity of CNTs and MXenes. Both nanomaterials effectively detect various pollutants in food, biological substances, and human bodies, making them invaluable in environmental monitoring and medical diagnostics. In conclusion, CNTs work better for biosensors that must be strong, flexible, and long-lasting under different conditions. MXenes, on the other hand, work better when chemical flexibility and compatibility with wet environments are essential.

Growing a Domestic Workforce for Oyster Aquaculture Through Operation of a Demonstration Oyster Farm in Maryland

The oyster aquaculture industry presents employment opportunities for a range of skill sets, education levels, and backgrounds (Engle, 2021), including but not limited to farm owners/operators, farm workers, food safety specialists, engineers, regulators, and researchers. Labor challenges pose an obstacle to growth, driven in part by lack of awareness of job opportunities in domestic aquaculture and lack of a trained workforce prepared to drive the industry forward (Sandolo, 2021).

The Influence of Storage Temperature and Packaging Technology on the Durability of Ready-to-Eat Preservative-Free Meat Bars with Dried Plasma.

Food business operators must include the results of shelf life testing in their HACCP plan. Ready-to-eat preservative-free meat products enriched with blood plasma are an unfathomable area of research in food safety. We tested modified atmosphere (80% N2 and 20% CO2) and vacuum packaged RTE preservative-free baked and smoked pork bars with dried blood plasma for Aerobic Plate Count, yeast and mould, lactic acid bacteria, Staphylococcus aureus, Enterobacteriaceae, Escherichia coli, and Campylobacter spp., and the presence of Listeria monocytogenes and Salmonella spp. during storage (temperatures from 4 to 34 °C) up to 35 days after production. The obtained data on the count of individual groups of microorganisms were subjected to analysis of variance (ANOVA) and statistically tested (Student's t-test with the Bonferroni correction); for temperatures at which there were statistically significant differences and high numerical variability, the trend of changes in bacterial counts were visualised using mathematical modelling. The results show that the optimal storage conditions are refrigerated temperatures (up to 8 °C) for two weeks. At higher temperatures, food spoilage occurred due to the growth of aerobic bacteria, lactic acid bacteria, yeast, and mould. The MAP packaging method was more conducive to spoilage of the bars, especially in temperatures over 8 °C.

European food safety research: An explorative study with funding experts’ consultation

The European research area exhibits considerable opacity and fragmentation in food safety research funding and organizational structures, impeding the exploitation of existing research potential across European countries. Given that food safety is inherently linked to the societal challenges of our time, identifying and removing existing barriers to research funding in this area is crucial. Towards investigating this matter, interviews were conducted with funding bodies from six European countries to assess key issues related to research funding in general and food safety in particular. Funding experts were then invited to a workshop to jointly discuss the challenges identified and explore strategies to address them. Evaluation of the food safety research funding situation in selected European countries revealed both convergences and significant differences among national funding bodies. Engaging with funding experts provided invaluable insights into the issues encountered with research funding, such as inadequate call management staff or insufficient research funds, culminating in a set of recommendations for action to remedy the situation.

Effect of Sonication Associated with Pasteurization on the Inactivation of Methicillin-Resistant Staphylococcus aureus in Milk Cream

Methicillin-resistant Staphylococcus aureus (MRSA) poses a significant challenge to the dairy industry, necessitating robust strategies to ensure food safety. This study focuses on the efficacy of thermosonication, a novel technology combining ultrasound and heat, in reducing MRSA in milk cream. Comparative analysis is conducted with conventional pasteurization, the industry standard. Results indicate that thermosonication effectively reduces MRSA counts by up to 4.72 log CFU/mL, akin to pasteurization’s reduction of 4.82 log CFU/mL. This finding highlights the potential of thermosonication as a rapid, energy-efficient alternative to pasteurization in the dairy industry, significantly reducing processing time while maintaining microbial safety. Further exploration and optimization of these techniques promise enhanced food safety and quality control in dairy products, addressing the growing concern of antibiotic-resistant strains like MRSA. This research lays a foundation for innovative approaches and underscores the significance of quantitative data in food safety research.

Design and synthesis of a novel fluorescent probe for the detection of bisulfite in plant and zebrafish

Sulfur dioxide (SO2) and its derivative bisulfite (SO32−/HSO3−) have a serious impact on the human health and environment. Therefore, the development of highly sensitive and specific detection methods for SO2 and its derivative is of great significance for environmental monitoring, food safety, and related disease research. In this paper, probe TPN-DP based on the triphenylamine fluorophore was developed to enable the imaging and detection of HSO3− in plant and zebrafish. In PBS buffer solution, the designed probe TPN-DP can react with HSO3− by highly selective nucleophilic addition reaction, leading to a sensitive change in the color of probe TPN-DP solution from pink to colorless. The changes in fluorescence and color can be easily recognized by the naked eye. Moreover, probe TPN-DP exhibited a fast response time of 8 min, high sensitivity of 19.4 nM, and excellent selectivity for sensing HSO3−. With the help of probe TPN-DP, the detection of HSO3− was successfully achieved in water sample and various biological systems, including onion epidermal cells, tobacco seedling roots, and zebrafish. These findings not only provide valuable insights into the biological role of HSO3− but also offer a promising analytical method for efficient HSO3− recognition.

Unveiling the potential of proteomics in addressing food and feed safety challenges.

The food and feed sector in Europe is rapidly evolving to address contemporary challenges, striving for fairer, safer, greener and more sustainable food systems. This includes the exploration of new protein sources for human consumption and animal feed such as protein derived from insects, algae or novel plant-derived proteins, and the re-evaluation of existing sources like processed animal protein (PAP). To generate reliable data on the diverse array of emerging protein sources for future food and feed safety assessments, a growing demand for the development and implementation of advanced analytical techniques exists. New approach methodologies (NAMs) including, mass spectrometry (MS)-based proteomics methods have been emerging as valuable techniques which potentially can be implemented in regulatory laboratory settings to complement conventional approaches in this realm. These MS-driven strategies have already proven their utility in diverse applications, including the detection of prohibited substances in feed, identification of allergens, differentiation of fish species in complex mixtures for fraud detection and the verification of novel foods and alternative protein sources. This EU-FORA programme was focused on three core objectives namely: (i) the training of the fellow in utilising MS-based proteomics for food and feed safety analyses, (ii) the involvement of the fellow in the development of standardised operating procedures (SOP) for targeted and non-targeted proteomic MS-based workflows for species and tissues specific PAP identification in a national reference laboratory (NRL) and (iii) the transfer and implementation of MS-based approaches and standardised protocols for PAP analysis at the fellow's home institution. Altogether, this programme facilitates the broadening and diversification of use of MS-based proteomic methodologies for reinforcing their significance within the domains of food and feed safety research and regulatory science applications.

Unveiling heterocyclic aromatic amines (HAAs) in thermally processed meat products: Formation, toxicity, and strategies for reduction - A comprehensive review.

This comprehensive review focuses on heterocyclic aromatic amines (HAAs), a class of chemicals that commonly form during the cooking or processing of protein-rich foods. The International Agency for Research on Cancer (IARC) has categorized certain HAAs as probable human carcinogens, highlighting the significance of studying their formation and control in food safety research. The main objective of this review is to address the knowledge gaps regarding HAAs formation and propose approaches to reduce their potential toxicity during thermal processing. By summarizing the mechanisms involved in HAAs formation and inhibition, the review encompasses both conventional and recent detection methods. Furthermore, it explores the distribution of HAAs in thermally processed meats prepared through various cooking techniques and examines their relative toxicity. Additionally, considering that the Maillard reaction, responsible for HAAs formation, also contributes to the unique flavors and aromas of cooked meat products, this review investigates the potential effects of inhibiting HAAs formation on flavor substances. A thorough understanding of these complex interactions provides a foundation for developing targeted interventions to minimize the formation of HAAs and other harmful compounds during food processing.