Microplastics (MPs) are a global concern due to their persistence in the environment and capacity to carry pollutants and pathogenic microorganisms. Given recent evidence on the co-occurrence of MPs and Campylobacter spp., the leading cause of foodborne gastrointestinal infections worldwide, this study investigates the role of MPs in Campylobacter jejuni contamination and their impact on antimicrobial susceptibility. The potential of five C. jejuni isolates from different origin (poultry, water, and human) to form biofilms on MPs over time (24 h, 48 h, and 72 h) was evaluated using traditional culture-dependent methods, including the reference strain C. jejuni subsp. jejuni strain NCTC 11168. The effect of MPs on the antimicrobial susceptibility of C. jejuni cells detached from MP-associated biofilms was also assessed using Etest strips. To comprehensively understand the interactions between the MPs and the bacteria, whole-genome sequencing was performed to explore the presence of adherence and biofilm-associated genes, as well as antibiotic resistance genes. The strains rapidly colonized the MPs within 24 h, exhibiting varying attachment densities over time ranging from 1.09 to 5.78 log CFU/MP, with strain NCTC 11168 identified as the strongest biofilm former overall. Furthermore, the abundance of adherence and biofilm formation genes was consistent with their abilities to form biofilm on MPs. The gene peb3 played a critical role in determining biofilm formation levels on MPs, while specific combinations of capA, capB, cj1725, and porA were linked to enhanced biofilm development. Similarly, the presence of antibiotic resistance determinants aligned with the phenotypic resistance, and only one strain (ST-6209/CC464) exhibited resistance to ciprofloxacin, nalidixic acid, and tetracycline. Notably, antimicrobial susceptibility of cells detached from MP-biofilms was increased by up to 4.6 log2-fold compared to planktonic counterparts. The findings indicate that MPs can facilitate the persistence of C. jejuni in the environment while simultaneously increasing their susceptibility to antibiotics. Further research with larger cohorts is needed to validate these preliminary observations in order to support the development of effective policies addressing MP pollution and food safety.

Micro- and nanoplastics (MNPs) represent an emerging class of contaminants, yet their biological impacts remain poorly understood. Mechanistic research comparing polymers such as polypropylene (PP) and polyethylene terephthalate (PET) is challenging because of strong differences in hydrophobicity, buoyancy or sedimentation behavior, size distribution (aggregation) and lack of strong acute cytotoxic effects. This study introduces an innovative in vitro dosing platform with 3D hepatic spheroids, providing a dosimetrically robust model to investigate metabolic and bioenergetic effects of chronic MNP exposure. Gel encapsulation was used to expose spheroids for up to 21 days to MNPs of different composition and size. MNPs accumulated in spheroids with polymer-specific localization. Cytotoxicity was limited, but metabolic profiling revealed consistent bioenergetic shifts. This work highlights the importance of particle size distributions and introduces alginate bead encapsulation as a reproducible dosing strategy for comparing MNPs and defining toxic doses linked to biologically relevant molecular and cellular events.

The food industry's focus on sustainability has driven efforts to replace multilayer packaging with recyclable monolayers. Monolayers face challenges in providing adequate gas barriers in modified atmosphere packaging (MAP), making oxygen's impact on food safety a critical concern. This study investigated the effects of oxygen on the surface growth of Listeria monocytogenes under refrigerated conditions. Both the absence and presence of mild acid stress were considered, with particular attention to strain-specific differences. Growth experiments were conducted using three L. monocytogenes strains (LMG 23905, LMG 23194 and ADQP 105) at 7 °C on brain heart infusion (BHI) agar under controlled oxygen (0 % and 21 %) and pH conditions (6.2 and 7.6). Significant differences in growth rates were observed for strain LMG 23905 under mildly acidic conditions (pH 6.2) at 0 % oxygen versus 21 % oxygen (p = 1.99e-13). Under neutral conditions (pH 7.6) however, no significant differences were observed (p = 0.09). Strain ADQP 105 exhibited no oxygen sensitivity under both mildly acidified (p = 0.1), and neutral pH conditions (p = 0.52). The same holds for strain LMG 23194, having identical growth rates under air and 100 % nitrogen at neutral (p = 0.28) and mildly reduced pH (p = 0.38). These findings highlight the strain-dependent effects of oxygen on the surface growth of L. monocytogenes. This also demonstrates that there is interaction between mild acid stress and oxygen availability, offering important insights for improving shelf-life predictions and better reflecting real-world conditions for bacterial contamination in solid foods.

Metabolic adaptation enables cancer cells to persist under fluctuating nutrient conditions and is a major driver of tumor progression, including colorectal cancer (CRC). The contribution of diet, such as red and processed meat, in tumor progression remains poorly understood. Here, we investigated how repeated exposure to hemin (HEM) and kynurenine (KYN), alone and in combination, shapes CRC cell metabolism under nutrient deprivation. Using respirometry and transcriptomics in 3D spheroids, we found that HEM and KYN induced distinct, context-dependent metabolic adaptations, differentially affecting glycolytic and oxidative energy pathways depending on nutrient availability. Co-exposure to HEM + KYN maintained energy production during glucose deprivation through enhanced lipid storage, revealing a mechanism by which red meat–derived metabolites reinforce metabolic plasticity. These findings demonstrate that red meat–derived metabolites modulate key bioenergetic pathways in a complementary manner and that their combined presence amplifies cancer cell adaptability, highlighting a potential mechanism linking diet to CRC progression.

Aflatoxin M1 (AFM1) is a carcinogenic milk contaminant and a persistent food safety concern in Serbia, especially under changing climate conditions that exacerbate contamination risks. This review synthesizes national research conducted between 2012 and 2024, covering more than thirty thousand analyzed milk and dairy samples, to evaluate AFM1 contamination, public health risks, and the need for structured risk ranking and prioritization frameworks recommended by the Food and Agriculture Organization (FAO) and the European Food Safety Authority (EFSA). A systematic analysis of Serbian studies explored AFM1 occurrence, dietary exposure, and health risk estimates across population groups. The evidence reveals persistent AFM1 contamination with pronounced seasonal peaks during drought years and winter months, frequently exceeding the EU maximum limit of 0.05 µg/kg. Recent multi-year studies confirm that climate-driven AFB1 contamination in maize and compound feed remains a significant and recurring source of AFM1 in milk, highlighting the necessity of structured risk prioritization frameworks. Exposure assessments highlight children and students as the most vulnerable groups, displaying the highest estimated daily intake. Although current margin of exposure (MOE) values remain within acceptable limits, the persistence of contamination underscores a need for proactive risk management. Adoption of FAO and EFSA risk-ranking methodologies would enhance monitoring efficiency, protect high-risk populations, and support alignment with EU standards. Implementing structured risk prioritization is crucial for strengthening Serbia’s food safety governance, guiding policy decisions, and reducing the health burden of AFM1 in the dairy sector.

Microplastics (MPs) are ubiquitous in aquatic ecosystems, where they are colonized by microbial communities, called the plastisphere. Of great concern is the detection of potential pathogens and antimicrobial resistance genes (ARG) in the plastisphere, which might be transported across ecosystems through MPs drifting. We used shotgun metagenomic profiling to assess taxa diversity, ARG and virulence genes (VG), within biofilm formed on polypropylene (PP) particles incubated in situ in five locations, following an anthropic gradient around Ostend (Belgium). Our results demonstrated significant variability of the plastisphere across incubation sites, but not between PP and control glass beads. Potential pathogenic bacteria (PPB) represented about 7% of bacterial reads within biofilms and VG were mainly involved in nutrition and adherence. Using dqPCR results to normalize metagenomic reads, we demonstrated a selective enrichment of ARG and VG in biofilms, while these were less abundant but more diverse in surrounding water. These findings highlight the presence of PPB, ARG and VG across all sites, likely driven by anthropogenic pressures. Although no substrate-specific effect was detected, the ability of PP particles to act as microbial reservoirs, coupled with their high mobility, reinforces concerns about their potential role in the transport and dissemination of microbial hazards.

Duchenne muscular dystrophy (DMD) is an X-linked neuromuscular disorder characterized by progressive muscle degeneration resulting in severe muscle weakness and motor function loss. Historically, therapeutic innovation has been facilitated by research in the mdx mouse, which is the most commonly used model for studying DMD. However, the inherent limitations of this model necessitate the formulation of alternative strategies. The present study proposes the zebrafish as an additional pre-clinical disease model in which to evaluate the benefits of nutraceuticals for DMD. The focus of this study will be taurine, a non-protein building amino acid that has emerged as a promising disease-modifying supplement for muscle wasting conditions, including aging-associated sarcopenia and DMD. The present study will evaluate the potential of early taurine supplementation to enhance the efficiency of autophagy and to improve mitochondrial health in a zebrafish DMD model. The study will facilitate a more rigorous examination of the claims pertaining to the beneficial effects of taurine on DMD progression, thereby informing the decision-making process regarding its potential implementation in human disease management.

Low water activity ingredients, including dried fruits, vegetables, herbs, and spices, are widely used in ready-to-eat (RTE) and non-RTE foods. Although low water activity limits microbial growth, bacterial pathogens can survive for extended periods in these products. Pathogen virulence, incorporation of dried ingredients in foods, and subsequent handling of the finished foods can collectively contribute to foodborne illness risk. This review summarizes the microbiological hazards associated with dried ingredients and discusses the effectiveness of conventional and emerging decontamination treatments for reducing pathogen levels. To support the evaluation of dried ingredients for RTE and non-RTE food applications, a qualitative risk-based microbiological decision framework is presented, along with practical examples demonstrating its use to rank risks and identify appropriate mitigation strategies. The framework considers the ingredient supply chain from production and harvest through packaging of finished products and integrates available outbreak, pathogen prevalence and levels, and product-specific data. This framework is intended to support the food manufacturing and food service sectors by providing a transparent, structured approach for evaluating, managing, and communicating pathogen risks associated with dry ingredients.

Risk assessment is a dynamic and continuously evolving process aimed at characterizing the potential adverse effects on life and health arising from exposure to hazards. Among these hazards, mycotoxins represent one of the most significant and widely investigated contaminants in food and feed. This review aims to summarize current knowledge on mycotoxin risk assessment practice, both for single substances and for cumulative risk assessment, and to provide practical guidance for future researchers. As individual substances, mycotoxins are well characterized from a toxicological perspective, and numerous risk assessments have been conducted globally across a wide range of food products. Although well-established methodological frameworks exist to support the cumulative risk assessment of mycotoxins, further efforts are needed to define common assessment groups, especially considering the pleiotropic nature of these compounds.

Bacillus cereus sensu lato is frequently involved in foodborne toxico-infections and is found in various foodstuff. It is unclear whether certain strains have a higher affinity for specific food matrices, which can be of interest for risk assessment. This study reports the characterization by whole-genome sequencing of 169 B. cereus isolates, isolated from 12 food types and soil over two decades. Any potential links between the food matrix of isolation, the isolate's genetic lineage and/or their (putative) virulence gene reservoir were investigated. More than 20 % of the strains contained the genes for the main potential enterotoxins (nheABC, hblCDA and cytK_2). Cereulide biosynthesis genes and genes encoding hemolysins and phospholipases, were detected in multiple isolates. Strain typing revealed a high diversity, as illustrated by 84 distinct sequence types, including 26 not previously described. This diversity was also reflected in the detection of all seven panC types and 71 unique virulence gene profiles. Core-genome MLST was used for phylogenomic investigation of the entire collection and SNP-based clustering was performed on the four most abundant sequence types, which did not reveal a clear affinity for specific B. cereus lineages or (putative) virulence genes for certain food matrices. Additionally, minimal genetic overlap was observed between soil and foodborne isolates. Clusters of closely-related isolates with common epidemiological metadata were detected. However, some isolates from different food matrices or collected several years apart were found to be genetically identical. This study provides elements that can be used for risk assessment of B. cereus in food.

Recent studies have detected microplastics (MPs) in seafood and various food products worldwide, including poultry, fish, salt, beverages, fruits, and vegetables. This widespread contamination makes human exposure through consumption unavoidable and raises concerns for food safety and human health. MPs provide physical support to microorganisms for biofilm formation, protecting them from extreme conditions and facilitating their persistence in the environment. However, little is known about the impact of MPs in the transmission of foodborne pathogens and subsequent spread of infectious diseases like campylobacteriosis, the most common foodborne illness caused by a bacterium, Campylobacter. This review explores the sources of MP contamination in the food chain and offers a comprehensive overview of MP presence in animals, food products, and beverages. Moreover, we compile the available studies linking MPs and Campylobacter and examine the potential impact of these particles on the transmission of Campylobacter along the food chain with a particular focus on poultry, the main source and reservoir for the pathogen. While the environmental and toxicological effects of MPs are increasingly understood, their influence on the virulence of Campylobacter and the spread of antimicrobial resistance remains underexplored. Further studies are needed to develop standardized methods for isolating and identifying MPs, enabling comprehensive investigations and more effective monitoring and risk mitigation strategies.

Fresh-cut iceberg lettuce is gaining popularity for its convenience, but it is highly susceptible to microbial spoilage. This study aimed to investigate the microbial community dynamics in commercially available fresh-cut iceberg lettuce packaged under controlled atmosphere in Belgium during storage at 7 °C for up to 12 days. To achieve a comprehensive understanding of the spoilage microorganisms and their interactions, a culture-complemented metataxonomic approach was conducted, including plating on non-selective and selective culture media for enumeration of total psychrotrophic counts (TPC), enterobacteria, Pseudomonas spp., psychrotrophic lactic acid bacteria (LAB) and yeasts. Throughout storage, TPC remained the predominant group, followed by Pseudomonas spp. until day 2, and by enterobacteria thereafter. The initial oxygen present in the commercial lettuce bags (1.5-2.0 %) was rapidly consumed within the first 24 h of storage (<0.1 %), resulting in a decrease in the dominance of Pseudomonas spp. (38.9 %) since then. By the end of storage, Pseudomonas spp. represented a minor group (<2 %), yet retained the capacity to grow. In contrast, Lactococcus spp. increased in abundance as CO2 rose (1.1-25.8 %), eventually dominating the bacterial community (>44 %) together with Serratia (21 %) and Rahnella (9.5-11 %) species. In fact, LAB showed the highest increase in population throughout storage, with levels rising from ∼1.8 log on day 0 to ∼7.5 log colony forming units per gram on day 12. These findings offer new insights into the deterioration mechanisms of fresh-cut lettuce stored under controlled atmosphere packaging, providing a basis for developing new strategies to prevent lettuce spoilage and thereby help the industry and retailers reduce food waste and associated economic losses.

An accurate, sensitive, low-cost, portable, and easy-to-use method for the quantitative detection of Clostridium perfringens (Cp), a zoonotic pathogen widely found in nature and capable of spreading through contaminated food or environments, is essential for epidemiology, prevention, and diagnostics. Here, we have designed a smartphone-based electrochemical DNA biosensor, which utilizes carboxyl-functionalized multi-walled carbon nanotubes (COOH-MWCNTs) as the substrate, L-cysteine (L-Cys) as the linker, and a synergistic modification of the bioelectrode with gold nanoparticles (AuNPs) and polyamidoamine dendrimers (PAMAM). This design enables highly sensitive, low-cost, label-free, and portable detection of the cpb2 gene in Cp. The sensor not only enhances detection performance but also improves convenience and practicality. Differential pulse voltammetry (DPV) was used to monitor the electrochemical signal response to changes in target DNA concentration, enabling the tracking of the DNA hybridization process. Under optimized conditions, the biosensor exhibited a linear detection range from 10⁻14 to 10⁻⁸ M, with a detection limit of 1.5 fM. It demonstrated excellent selectivity for the cpb2 gene and was successfully applied to detect variations in cpb2 gene content in wastewater and fecal DNA samples. Compared with the traditional method, the detection time of this method is short, the operation of professional and technical personnel is not required, the instrument is small and portable, and the single detection cost is significantly reduced. This study provides a new strategy for the rapid, portable, and highly sensitive detection of bacterial toxin genes in livestock and aquaculture.

The potential health risks of consuming crops irrigated with microcystin-contaminated water are increasingly recognized. While previous studies demonstrated microcystin accumulation in crops, they do not fully reflect real-world conditions and have data gaps. Moreover, some available studies also obtained data with suboptimal analytical methods.Therefore, this study evaluated microcystin-LR (MC-LR) accumulation in strawberries, carrots, and lettuce under spray and drip irrigation, using a state-of-the-art, validated UHPLC-MS/MS method in a medium-scale setup similar to commercial greenhouse production.MC-LR was detected in the outer and middle leaves of lettuce after spray irrigation. In carrots, only a few replicates showed MC-LR in the greens or taproot, yielding inconclusive results. In strawberries, MC-LR accumulated in the roots, foliage, and fruit after spray irrigation. Additionally, drip irrigation led to MC-LR accumulation in strawberry roots and greens, even though only the roots were directly exposed.The findings suggest that strawberry plants are particularly susceptible to microcystin accumulation when irrigated with contaminated water, highlighting the need for risk mitigation measures.

Despite extensive investigation into the anti-cancer activity of the natural polyphenol curcumin, its therapeutic application is restricted by its inherent physicochemical properties. Synthetic curcumin analogues, however, offer a promising strategy to improve the drug-like potential of curcumin. In this study, we evaluated three curcumin-based benzothiazepane analogues for their ability to selectively target colon cancer cells. Their cytotoxicity was assessed on intestinal cancerous HCT-116 and non-cancerous IPEC-J2 cells using cell viability assays and microscopic imaging. Two analogues, AT007 and AT096, demonstrated enhanced anti-cancer selectivity compared to curcumin. Interestingly, this effect correlated with the aggregation of these compounds in cell medium, which was influenced by compound concentration and medium composition (particularly the presence of albumin). Confocal microscopy confirmed the presence of particles up to 12 µm inside both cell lines, yet downstream metabolic and transcriptomic responses revealed distinct coping mechanisms that may underlie the higher survival of IPEC-J2 cells. Rather than direct molecular interactions typical of soluble compounds, the observed selectivity appears to result from indirect, particle-driven physical effects, potentially involving (intracellular) membrane disruption. Our findings suggest that aggregation behaviour can be a key determinant in improving the potency and selectivity of bioactive compounds, opening new opportunities for the design and screening of more selective anti-cancer therapeutics.

Abstract The Scientific Committee is asked to address the following questions: – From a food safety point of view, can it be accepted to deviate from the legal storage temperature of food products that must be kept refrigerated, including poultry roulade, processed cheese and fried mushrooms, intended for the preparation of tacos? Specifically, may these food products be kept at a temperature of up to 30°C (instead of up to 7°C) for maximum 3 hours? – If a derogation can be granted, it is asked whether a deviation from the legal storage temperature is also acceptable for other food products, from the categories of deli meats, cheese and vegetables intended for taco preparation, without carrying out a full risk assessment for these additional food products. Salmonella spp., Shiga toxin‐producing E. coli (STEC), L. monocytogenes and B. cereus have been identified as the most relevant hazards in this context. Based on the available information (shelf‐life tests, challenge tests and simulations), the requested deviation (storage for 3 hours at maximum 30°C instead of the legal temperature of 7°C) is not acceptable in terms of food safety. Based on the simulations of the growth potential of microorganisms performed using the Combase interface, a number of time/temperature combinations are identified as acceptable for the food products concerned (cooked deli meats, pasteurized cheese and heated vegetables) during taco preparation.

Iminosugars have a carbohydrate-like backbone in which the ring oxygen is replaced by nitrogen. They are naturally found in foods such as rice, buckwheat, mulberries, and fermented vegetables, and are reported to exert anti-hyperlipidemic and anti-hyperglycemic effects due to the inhibition of cellular glycosidases. This mechanism suggests their potential role in cancer treatment and prevention. In this study, two natural iminosugars, D-fagomine (FGM) and 1-deoxynojirimycin (DNJ), and their synthetic derivatives were screened for potential anticancer properties using Caco-2 and HCT-116 cells as models for the early and late stages of colon cancer, respectively. Iminosugars were found to decrease cell viability, with effects varying based on the type of iminosugar, cell type, growth condition (glucose concentration), exposure time (1 vs. 13 days), and tissue architecture (monolayer vs. spheroid). The combined use of innovative techniques, such as IncuCyte® live cell imaging and Seahorse real-time cellular metabolic analysis, and microscopic observation after staining enabled us to detect changes in substrate utilization for energy metabolism, including increased glycolysis and alterations in lipid and glycogen stores. The evidence that iminosugars, both natural and synthetic, influence cellular bioenergetics paves the way for their potential use in various applications, including cancer treatment.

Food safety is a significant issue of global concern. Consumer safety and government regulations drive the need for the accurate analysis of food contaminants, residues and other chemical constituents of concern. Traditional methods for the detection of food contaminants often present challenges, including lengthy processing times and food matrix interference; they often require expensive equipment, skilled personnel or have limitations in sensitivity or specificity. Developing novel analytical methods that are sensitive, specific, accurate and rapid is therefore crucial for ensuring food safety and the protection of consumers. The immuno-polymerase chain reaction (IPCR) method offers a promising solution in the analysis of food contaminants by combining the specificity of conventional immunological methods with the exponential sensitivity of PCR amplification. This review evaluates the current state of IPCR methods, describes a variety of existing IPCR formats and explores their application in the analysis of food contaminants, including pathogenic bacteria and their toxins, viruses, mycotoxins, allergens, polycyclic aromatic hydrocarbons, polychlorinated biphenyls, phthalic acid esters, pesticides, antibiotics and other food contaminants. Depending on the type of analyte, either sandwich or competitive format IPCR methods are predominantly used. This review also examines limitations of current IPCR methods and explores potential advancements for future implementation in the field of food safety.

In Serbia, contamination of milk and dairy products with aflatoxin M1 (AFM1) represents a recurrent food safety challenge, reflecting the dairy chain's susceptibility to climate extremes, feed contamination, and improper storage conditions. Protecting public health, particularly that of children, requires systematic approaches that move beyond descriptive monitoring towards structured risk prioritisation. The aim of this study was to apply the FAO (2020) and EFSA (2015) methodologies for risk ranking and prioritisation to AFM1 in Serbia, thereby providing an evidence-based framework to guide risk-based monitoring and food safety governance. The analysis was based on data synthesised from peer-reviewed studies on AFM1 occurrence, dietary exposure, and risk characterisation in different dairy product categories. Risk matrices were constructed by combining likelihood of contamination with severity of health outcomes, expressed through Margin of Exposure (MOE) values. In addition, binomial probability principles were applied to design optimised sampling strategies. Results showed that raw and pasteurised/UHT milk represent the highest-priority categories, reflecting both high contamination frequencies and dietary importance for children. Average MOE values exceeded the health-based threshold, but high consumers, particularly toddlers, were at risk of falling below safe levels. Probability-based sampling demonstrated how resources can be rationally allocated to maximise detection assurance. This study concludes that structured risk ranking and prioritisation provide a transparent foundation for proportionate AFM1 monitoring and control.