Abstract Food chemical safety assessment is increasingly challenged by diverse exposure scenarios arising from environmental contamination, food processing, food-contact materials (FCMs), and emerging food systems. These challenges highlight the need for approaches with greater scalability, mechanistic resolution, and human relevance. This review examines how high-throughput toxicity testing (HTT), integrated with mechanistic interpretation and kinetic modeling, can contribute to next-generation risk assessment (NGRA) of food-relevant chemicals. We summarize advances in omics-based toxicology, phenotypic profiling, adverse outcome pathway (AOP) frameworks, benchmark dose (BMD) modeling, and physiologically based kinetic modeling (PBK) coupled with quantitative in vitro to in vivo extrapolation (QIVIVE). Case studies involving contaminants, additives, food-contact material migrants, nanomaterials, and novel foods illustrate how HTT can identify early molecular, cellular, and phenotypic perturbations and provide mechanistic evidence for hazard prioritization. AOP frameworks provide mechanistic context for interpreting HTT-derived bioactivity signals, while BMD modeling and PBK-QIVIVE enable quantitative translation toward human-relevant dose metrics. However, regulatory application remains limited by insufficient quantitative AOPs, dose–response uncertainty, mixture complexity, and data standardization challenges. Coupling HTT with mechanistic and kinetic modeling provides an integrative decision support framework to advance more human-relevant, transparent, and adaptive food chemical safety assessment.

The requirement of protospacer-adjacent motif (PAM) and nucleic acid amplification have restricted the application of CRISPR/Cas12a system in bacteria detection. Herein, we proposed a restriction endonuclease mediated, "one to more", PAM-independent and amplification-free CRISPR/Cas12a strategy for bacteria detection. The bacterial genomic DNA can be cleaved by specific endonuclease, exposing a large number of sticky ends. The combination of these exposed sticky ends with a short ssDNA could effectively activate the trans-cleavage activity of Cas12a to break a FQ reporter probe, achieving the purpose of detection. This approach not only overcomes the dependence of PAM sequence but also enables one-pot detection of target bacteria within 60 min without the requirement of nucleic acid amplification. The detection sensitivity of two typical food contaminated bacteria, Alicyclobacillus acidoterrestris and Cronobacter sakazakii was 13.38 CFU/mL and 18.67 CFU/mL, respectively. In summary, this work provided a novel strategy and powerful tool for bacteria analysis and molecular diagnosis.

A portable conjugated microporous polymer (THDH-DMA)-based fluorescent sensor coupled with a smartphone enables rapid, selective, and sensitive on-site detection of Fe3+ in water.

Antibacterial hydrogels represent a class of biomaterials that integrate multicomponent synergy and multimodal antibacterial mechanisms. Precise engineering of their composition and modular architecture enables the integration of potent antibacterial action, versatility, and enhanced biocompatibility, demonstrating remarkable potential for advanced antimicrobial applications. This review provides a comprehensive and critical analysis of cross-linking design paradigms, classification, antibacterial mechanisms, and applications of antibacterial hydrogels. We emphasize that elucidating structure-activity relationships and deeper understanding of antibacterial mechanisms are critical for developing novel antibacterial hydrogels, especially for food applications. The review culminates in identifying pivotal research directions and persistent technical challenges that must be addressed to propel the field forward.

The overuse or abuse of antibiotics drives the global health threat of antimicrobial resistance. Although bans on certain veterinary antibiotics, such as colistin, have proven effective, the impact of fluoroquinolone stewardship on the evolution of the foodborne pathogen Salmonella enterica serovar Enteritidis (S. Enteritidis) remains unclear. Here, we conducted a decade-long (2014-2023) retrospective longitudinal genomic epidemiological analysis of 441 ST11 S. Enteritidis isolates from Guangxi, China, alongside a global reference dataset of 4297 genomes. Our aim was to elucidate the effect of real-world antibiotic stewardship on the shift of gyrA point mutations and lineage distribution. Surveillance identified three global epidemic clade sublineages (GEC-L2, L3, L4), with the multidrug-resistant GEC-L4 (i.e., GC-c or MMC2), characterized by the gyrA mutation with amino acid substitution D87Y, being domestically dominant (70.07%, 309/441). Following China's 2016 ban on the veterinary use of critical fluoroquinolones, the proportion of the highly resistant GEC-L4 sublineage decreased continuously (from 86.84% in 2017 to 56.00% in 2023), while the less resistant GEC-L3 sublineage (i.e., GC-b or MMC1), mainly characterized by gyrA D87G, increased simultaneously (from 13.16% to 44.00%). This phenomenon might be attributed to the fact that the GEC-L4 sublineage exhibited a higher fitness cost compared with the GEC-L3 sublineage, as confirmed by the competition assay. A Random Forest Model validated that the gyrA mutation with amino acid substitution D87Y was the paramount feature for these sublineages' identification. In contrast, global data showed a continuous increase in gyrA mutations (from 8.63% in 2006 to 68.85% in 2024), primarily D87Y (from 1.44% to 31.15%) and D87N (from 4.32% to 22.95%), correlating with rising average fluoroquinolone consumption. This study provides direct genomic evidence that national-level antibiotic stewardship can drive the replacement of highly resistant sublineages with moderately resistant ones. These findings offer crucial scientific evidence for evaluating the impact of antibiotic management policies and inform strategies for the rational use of antimicrobials.

What is already known about this topic?:The increasing consumption of preprepared dishes in China has raised public health concerns regarding potential exposure to per- and polyfluoroalkyl substances (PFASs). What is added by this report?:Trifluoroacetic acid was the predominant PFAS in all samples, while long-chain PFASs were mainly detected in aquatic foods. A conservative scenario-based dietary exposure assessment showed that the estimated daily intake of total PFASs ranged from 1.14 to 98.63 ng/kg body weight/day across various preprepared dishes. What are the implications for public health practice?:Current PFASs exposure from preprepared meat and aquatic dishes poses a low health risk for consumers, but continued monitoring and risk assessment remain necussary.

DNA repair intermediates such as abasic (AP) sites are among the most abundant and reactive DNA lesions. Despite their central roles in repair fidelity and as therapeutic targets, their transient dynamics and covalent reactivity remain difficult to probe with existing methods. Here, we introduce a molecular anchor-assisted strategy that immobilizes AP-containing ssDNA within the α-hemolysin (α-HL) nanopore, enabling real-time monitoring of its reactivity dynamics. Vitamin B12 is covalently attached to thiol-modified ssDNA through a cobalt-sulfur bond, serving as a molecular anchor to tether ssDNA inside the α-HL nanopore. The resulting B12-anchored ssDNA produces uniform current blockages of defined amplitude. This approach allows high-resolution discrimination of the four canonical nucleobases and dynamic tracking of AP site formation. By recording the reactions of hydroxylamine compounds with AP sites, we discovered that different substituent groups on hydroxylamines modulate their reactivity toward AP sites. Furthermore, this B12-anchored ssDNA strategy functions as a single-molecule reactor, enabling the capture of transient chemical structural changes at AP sites and direct observation of their reaction processes with nucleophilic agents at the single-molecule level. The results demonstrate that nanopore-based single-molecule profiling enables detailed kinetic and mechanistic studies of reactive DNA intermediates, which are otherwise difficult to monitor, thereby highlighting its potential for molecular diagnostics and targeted therapeutic development.

Objective Hospital wastewater serves as a critical source of antimicrobial resistance genes (ARGs), particularly those associated with opportunistic pathogens like Raoultella ornithinolytica. This study aimed to characterize carbapenem-resistant R. ornithinolytica (CR-ROR) isolates carrying the tmexCD2-toprJ2 gene cluster from hospital sewage in Zhejiang province, China. Methods Wastewater samples collected from four hospitals in Zhejiang Province were screened for carbapenem-resistant isolates. Whole-genome sequencing using the Illumina platform, combined with bioinformatic analysis, was performed to investigate phylogenetic relationships, associated ARGs, and the structural features of plasmids in tmexCD2-toprJ2-positive CR-ROR isolates. Results Five genetically distantly related tmexCD2-toprJ2-positive CR-ROR isolates were identified. All exhibited multidrug-resistant (MDR) phenotypes and carried various ARGs, including carbapenemase genes such as blaKPC-2, blaNDM-1, and blaIMP-4. Genomic analysis revealed that tmexCD2-toprJ2 was plasmid-borne and frequently flanked by mobile genetic elements (MGEs), suggesting a high risk of horizontal gene transfer. Conclusions The presence of tmexCD2-toprJ2-carrying CR-ROR in hospital sewage underscores the role of wastewater as a potential environmental reservoir for clinically resistant genes. Ongoing surveillance of hospital effluents is crucial for tracking the dissemination of high-risk antimicrobial resistance determinants and informing timely public health interventions.

Given the serious harm that microcystins (MCs) cause to public health and ecosystems (e.g., freshwater systems), there is a great need for an on-site MCs environmental assay that is cost-effective, easy-to-use, reliable and precise. Regarding this point, we here proposed a molecular imprinting-based selectively regulated laccase-like copper-doped carbon dot nanozyme (MIP@Cu-CD) to engineer a smartphone-assisted portable device, as a proof of concept, yielding the on-site visual colorimetric/fluorescence imaging-driven precise and reliable detection of MC-LR (a representative MCs) in real environmental waters. The detection mechanism was derived from the combination of MC-LR with its recognizable molecularly imprinted cavity, blocking the laccase-like activity-triggered MIP@Cu-CD to oxidize 2,4-dichlorophenol (2,4-DP) into a red quinoneimine product in the presence of 4-aminophenol (4-AP). At this moment, a decrease in colorimetric absorbance at 510 nm and a recovery of intrinsic blue fluorescence at 420 nm were observed in the MIP@Cu-CD + 2,4-DP + 4-AP system, with an increase in the MC-LR concentration. On this basis, the system of MIP@Cu-CD + 2,4-DP + 4-AP was integrated with a home-made smartphone-assisted portable device to perform on-site visual colorimetric/fluorescence image-based RGB detection of MC-LR in tap water, urban sewage, and lake water, with detection limits of 0.227 mu g L-1 and 0.101 mu g L-1 in colorimetric and fluorescence ways, respectively. Moreover, the reliability was also confirmed by the liquid chromatography-mass spectrometry (LC-MS) method. This work provides a feasible approach to utilize a molecular imprinting-based laccase-like nanozyme strategy to integrate portable devices for realizing the on-demand on-site detection of MCs in environmental waters.

Seafood is primary source of mercury exposure for general population. However, the health risks related to seafood mercury was fail to be accurately evaluated due to lack of applicable bioavailability on Hg species. We herein comprehensively evaluated the in vitro and in vivo bioavailability of Hg species in various seafood matrices, including seaweed, fish and shellfish. Results revealed that in vitro bioavailability determined using Caco-2 cells seem to vary along with both seafood’s matrices and Hg species. Whereas, in vivo bioavailability obtained using mouse exposure seems primarily dependent on Hg species rather than seafood’s matrices. In vitro bioavailability of Hg2+ in all types of seafood is higher than that of methyl mercury (MeHg) and ethyl mercury (EtHg), while MeHg and EtHg in the same type of seafood have similar values. In contrast, in vivo bioavailability seem to exhibit an opposite variation trend, with MeHg/EtHg having higher values than that of Hg2+ in all seafood matrices. Results indicated that Caco-2 cells model might not serve as a direct alternative to animal model for accurately evaluating bioavailability of mercury, especially MeHg/EtHg, in seafood. Simulated calculations of target hazard quotient indicated that accurate detection of Hg species contents and acquisition of their reliable bioavailability are crucial for accurately evaluating the seafood mercury-related health risks. Results of this study offered reference-worthy bioavailability data for Hg species in seafood, which facilitates the accurate evaluation of mercury-related health risks from seafood consumption, and offered valuable insights into bioavailability evaluation of mercury in other foods.

Ionic liquids (ILs) are increasingly promoted as sustainable solvents, yet their structural stability and hydrophobicity raise concern that some may behave as PFAS-like emerging contaminants. Here, we investigated the toxicokinetics of three alkylimidazolium ILs (AM-ILs; C6[MIM], C8[MIM] and C10[MIM]) in rats using a validated UPLC-MS/MS integrated with physiologically based toxicokinetic (PBTK) modeling. Plasma profiles revealed rapid oral absorption (Tmax 0.29–0.83 h) and dose-dependent increases in systemic exposure. At equal dose, the area under the curve decreased with increasing alkyl chain length, whereas elimination half-lives increased, indicating enhanced persistence with longer alkyl substituents. Application of the plasma-validated PBTK model to the liver and kidney predicted that AM-ILs organ burdens increased with both dose and alkyl chain length, closely mirroring the distribution and retention patterns observed for long-chain PFASs. These findings highlight that AM-ILs exhibit rapid systemic uptake, chain-length-dependent toxicokinetics, and organ-specific accumulation, conferring potential for chronic internal exposure and bioaccumulative risk. In this study, a multi-chain length PBTK model was developed and validated for the first time, integrating systemic toxicokinetic data with organ-specific accumulation characteristics, underscoring the necessity of incorporating ILs into emerging contaminant assessment frameworks for long-term risk evaluation.

Nanoplastics (NPs) are common hydrophobic contaminants in food packaging, readily interacting with other pollutants to produce severe unknown toxicity. Heterocyclic amines (HAAs) are carcinogenic hydrophobic byproducts commonly generated during the thermal processing of meat products. With the widespread adoption of takeout food, coexposure risks to NPs and HAAs are inevitable, yet studies on their combined contamination remain scarce. This study comprehensively evaluated the chronic toxic effects of coexposure to polystyrene nanoplastics (PS-NPs) and 9H-pyridine[3,4-b]indole (Norharman) on zebrafish (Danio rerio) through behavioral, histopathological, and transcriptomic analyses. Findings reveal that both PS-NPs and Norharman induce behavioral abnormalities with significant synergistic effects. This interaction likely disrupts the blood-brain barrier (BBB) by downregulating Occludin gene expression, triggering oxidative stress and apoptosis in the brain, and activating neuroinflammatory responses mediated by the MAPK signaling pathway. This study reveals the synergistic neurotoxic mechanism of coexisting food contaminants, providing a theoretical basis for assessing combined exposure risks.

Sensitive, rapid, and reliable detection of aflatoxin B1 (AFB1) in food is of great importance for ensuring food safety. In this study, a dual-mode paper-based biosensing platform was developed by integrating the CRISPR/Cas12a nucleic acid cleavage system with a nanozyme-responsive DNA hydrogel for highly sensitive and portable detection of AFB1. Upon the presence of the target, AFB1 binds specifically to the aptamer, thereby activating the CRISPR/Cas12a system and subsequently inducing the degradation of the hydrogel network, which leads to the release of the encapsulated Au@Pt nanozyme. The released nanozyme catalyzes the oxidation of TMB in the presence of H2O2 to generate a blue-colored product, while simultaneously producing a quantifiable diffusion signal on the paper-based substrate, enabling dual-mode colorimetric and distance-based detection. The detection limit of naked-eye detection is 0.1 ng mL−1, and it is 0.075 ng mL−1 in the distance detection mode. The proposed method can complete the signal output within 30 min and exhibits the advantages of operational simplicity, rapid response, and high sensitivity. In practical peanut samples, satisfactory recoveries and good stability were achieved, demonstrating its potential as a promising strategy for on-site detection of trace mycotoxins in food.

The contamination of organophosphate esters (OPEs) has increased sharply in China due to the regulation of brominated flame retardants (BFRs) in recent years; however, data on their occurrence in humans are limited. In this study, fourteen OPEs and ten of their metabolites (mOPEs) were measured in pooled human milk samples collected across China between 2016 and 2019. Both OPEs and mOPEs were ubiquitous in the tested 100 samples, with median levels of 2.2 and 1.05 ng/mL, respectively. OPE levels had significantly surpassed those of brominated flame retardants (BFRs) in the tested samples, indicating that the consumption pattern of flame retardants in China has shifted from BFRs to OPEs. The significant correlations between some mOPEs and their parent OPEs, and the relative abundance of mOPEs, suggested that certain OPEs were inclined to transform to mOPEs, while a portion of mOPEs may directly originate from external exposure. The present human milk survey was conducted at the same time and site as the 6th China total diet study, and the intakes of some OPEs through food consumption among lactating women were significantly and positively correlated with their concentrations in human milk, indicating that food consumption might be a main pathway for human exposure to OPEs. The estimated daily intakes (EDI) of OPEs/mOPEs via breastfeeding for the Chinese infants were calculated. Under medium exposure, the EDIs of ∑14OPEs/∑10mOPEs for infants aged 0-6 months, 7-12 months, and 13-24 months were 325/148, 197/89.7, and 139/63.4 ng/kg bw/day, respectively, and the daily intake of OPEs/mOPEs via breastfeeding might not cause significant health risks. However, since most OPEs and all mOPEs lack RfD due to limited toxicology data, the health risks posed by exposure to both OPEs and mOPEs should be continually evaluated.

ObjectiveThis study aims to investigate the public’s cognition and characteristics regarding prepared dishes.MethodsThis study systematically collected 707 155 public comments related to “prepared dishes” on online platforms from May 2023 to June 2024， followed by descriptive and structural equation modeling analysis.ResultsThe overall attitude of the public tended to be negative， with a satisfaction rate of 31.50% and a purchase intention rate of 36.76%. The main negative emotions were doubt and blame， and the average cognitive depth was 1.48， which was at a relatively low level. Among different categories， prepared vegetable dishes were more difficult to be accepted. The public’s willingness to purchase prepared dishes was mainly influenced by factors such as price， nutritional content of raw materials， and quality characteristics， which is consistent with the current public opinion situation.ConclusionRelevant government departments should pay attention to the public perceptions and public opinion patterns regarding prepared dishes， optimize targeted risk management measures and better address consumer needs.

Taurine, an important nutritional fortifier in infant formula, is closely associated with neurodevelopment and immune homeostasis. However, existing studies have mainly focused on its immediate promoting effects on neurodevelopment, while whether early-life taurine supplementation confers long-term neuroprotective effects during later developmental stages remains unclear. In this study, a three-stage zebrafish model was established, including early-life taurine supplementation, cessation of supplementation, and subsequent exposure to acrylamide during later development, to evaluate the long-term neuroprotective potential of taurine. The results showed that early-life taurine supplementation significantly alleviated neurobehavioral deficits, oxidative damage in brain tissue, and inflammatory responses induced by acrylamide exposure during later developmental stages. Mechanistically, taurine may exert these effects through coordinated regulation of neuroimmune and metabolic pathways. At the immune level, taurine inhibited NF-κB-related inflammatory activation, suppressed M1-associated microglial markers, and promoted M2-associated responses, thereby improving the neuroimmune microenvironment. At the metabolic level, taurine regulated FOXO pathway-related gene expression and improved glycolysis-related metabolic disturbance, which was associated with reduced oxidative damage and improved energy homeostasis. Overall, these findings suggest that transient taurine supplementation during early life may reduce later susceptibility to foodborne neurotoxicant exposure, which is associated with nutritional programming-like changes in neuroimmune and metabolic pathways. This study provides a new perspective for early-life nutritional intervention and dietary risk assessment.

Argonautes (Agos) are next-generation programmable nucleases that demonstrate great potential for the detection of nucleic acid targets. However, a contemporary difficulty in the intensely active field of Ago-based biosensing is how to enhance its flexibility to meet the requirement in versatile non-nucleic acid target detection. Herein, we proposed an Ago enzymatic activation state regulation strategy to achieve diverse target detection. The normal linear guide DNA of Ago is confined in a hairpin conformation, termed the hgDNA. This unique configuration could effectively prevent Clostridium butyricum Ago (CbAgo) activation. Under the action of enzymatic hydrolysis or small-molecule-mediated attack on specific sites, the hairpin structure of hgDNA is disrupted, leading to the release of active gDNA. Therefore, the activation and inactivation states of CbAgo can be deliberately regulated. The possible activation mechanism could be ascribed to the strand displacement reaction between unstable hgDNA and the complementary strand. This tunability transforms CbAgo into a generalizable detection platform suitable for various targets such as nucleases, small molecules, and pathogens. This study advances the dynamic response of CbAgo to gDNA conformations and introduces the universal concept of Ago-based non-nucleic acid detection to broaden the next-generation biosensing applications.

Zearalenone (ZEN), a mycotoxin present in cereals, poses significant health risks to animals and humans due to its estrogenic effects. Numerous studies on the enzymatic detoxification of ZEN have predominantly focused on reducing the parent toxin to assess the enzyme's efficacy, yet there is limited research on the identification and toxicity evaluation of the enzymatic degradation products. This study investigated the enzymatic degradation mechanisms of ZEN using commercial peroxidase (POD) and laccase (LC), with a focus on identifying degradation products and assessing their hepatotoxicity effects. Molecular docking and dynamics simulations elucidated the binding mechanisms between these enzymes and ZEN, revealing strong interactions that facilitate efficient detoxification. Subsequent analysis employing ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS) successfully identified crucial degradation products. Hepatic toxicity of the enzymatic degradation products was comprehensively assessed in HepaRG liver cells through systematic measurements of cell viability, oxidative stress, apoptosis, mitochondrial membrane potential, and molecular metabolic profiles. Our findings demonstrate that both POD and LC exhibit significant efficacy in mitigating hepatocyte toxicity induced by ZEN, thereby highlighting their potential utility in enhancing food safety. This research provides essential data for safety evaluation regarding enzymatic detoxification of ZEN while offering theoretical and technical resources for risk assessment related to mycotoxin enzymatic detoxification.

Perfluorooctanoic acid (PFOA) in soil poses a significant threat to reptile health. Given the global decline in reptile abundance and diversity, understanding reptile responses to prolonged PFOA-contaminated soil is urgently needed. In this study, Eremias argus, small lizards distributed across northern China, were exposed to PFOA (0.05 and 0.5 mg/kg) for 60 days. PFOA burden, physiological parameters, histopathological features, untargeted metabolomics, and 16S rDNA sequencing were performed to investigate the effects of soil PFOA exposure. The results revealed pronounced sex-dependent differences in PFOA accumulation and associated biological responses. Males exhibited significantly higher internal PFOA levels, whereas females showed comparatively lower burdens, which may be associated with maternal transfer of PFOA to eggs. These sex-dependent differences were accompanied by distinct pathological and metabolic alterations. Specifically, males exhibited severe intestinal barrier damage, reduced abundance of Bacteroidetes, and metabolic disturbances related to bile acid homeostasis. In contrast, females maintained relatively intact intestinal architecture and more stable Bacteroidetes populations, together with comparatively stable bile acid-related metabolic profiles, which potentially facilitating contaminant excretion. Overall, these findings provide mechanistic insights into how soil PFOA exposure may affect reptile physiology during sensitive life-history stages, thereby offering relevant information for future ecological risk assessments.

Food safety systems are generally effective for known contaminants but are less prepared for chemical hazards that are not captured through routine monitoring. This challenge is particularly relevant for animal-derived foods, where hazardous chemicals may enter and change along the production chain, hindering their detection and interpretation. This review discusses the origins of emerging chemical hazards in animal-derived foods, why they can be overlooked by conventional monitoring, and strategies for integrating detection with public health frameworks. Targeted methods remain essential but are less capable of capturing low-concentration, transformed, or unexpected compounds. When combined with suspect screening and confidence-based identification, high-resolution mass spectrometry can extend detection beyond predefined targets and improve the interpretation of unexpected chemical signals. For public health practice, the value of expanded detection lies not only in identifying more chemicals but also in determining which signals require confirmation, evaluation, and consideration for future monitoring and control.