The ubiquitous presence of antimicrobial-resistant organisms and antimicrobial resistance genes (ARGs) constitutes a major threat to global public safety. Tetracycline (TET) is a common antimicrobial agent that inhibits bacterial growth and is frequently detected in aquatic environments. Although TET may display coselection for resistance, limited knowledge is available on whether and how it might influence plasmid-mediated conjugation. Subinhibitory concentrations (3.9–250 ng/mL) of TET promoted horizontal gene transfer (HGT) via the mobilizable plasmid pVP52-1 from the donor Vibrio parahaemolyticus NJIFDCVp52 to the recipient Escherichia coli EC600 by 1.47- to 3.19-fold. The transcription levels of tetracycline resistance genes [tetA, tetR(A)], conjugation-related genes (traA, traD), outer membrane protein genes (ompA, ompK, ompV), reactive oxygen species (ROS)-related genes (oxyR, rpoS), autoinducer-2 (AI-2) synthesis gene (luxS), and SOS-related genes (lexA, recA) in the donor and recipient were significantly increased. Furthermore, the overproduced intracellular ROS generation and increased cell membrane permeability under TET exposure stimulated the conjugative transfer of ARGs. Overall, this study provides important insights into the contributions of TET to the spread of antimicrobial resistance.

V. parahaemolyticus is one of the most important foodborne pathogens and poses a serious threat to public health because of its high incidence and transferable antimicrobial resistance genes (ARGs). In this study, a novel approach for simultaneous detection of V. parahaemolyticus and the tetracycline resistance gene tetA was developed by reasonably integrating rapid loop-mediated isothermal amplification (LAMP) with the powerful cleavage ability of Pyrococcus furiosus Argonaute (PfAgo). The guide DNAs (gDNAs) were carefully designed to complete continuous signal transduction. With this strategy, the dual-target LAMP amplicons that were originally difficult to distinguish could be ingeniously converted into fluorescent signals. The duplex LAMP-PfAgo method was used to complete the entire process within 45 min at constant temperature and showed no cross-reactivity with non-target pathogens. The results demonstrated superior analytical performance, with a limit of detection (LOD) of as low as 6.68 CFU/mL for the V. parahaemolyticus carrying the tetA gene and a dynamic range from 10° to 10⁶ CFU/mL. Furthermore, the proposed method exhibited satisfactory sensitivity and feasibility for use in identifying contaminated samples (water, salmon and oysters). In summary, this study provides effective technical support for the on-site detection of pathogenic bacteria and tetracycline resistance genes, presenting promising prospects for food safety testing, clinical diagnosis, and environmental monitoring in the One Health context.

A paper-based surface enhancement of a Raman scattering substrate consisting of silver-nanowires stacked on glass-fiber filter paper was prepared. At the same time, the DNA-embedding molecule Eva Green was introduced as a signaling molecule for surface-enhanced Raman scattering (SERS) detection. Polymerase chain reaction (PCR) was used to amplify target genes and the method was developed into a rapid molecular diagnostic system. The total detection time of the developed detection method was 40 min, including 30 min of PCR amplification and 10 min of SERS measurement. After 30 PCR cycles, bacterial DNA with an initial concentration of 20 fg/μL and a bacterial suspension with an initial concentration of 7.2 × 10¹ CFUs/mL could be detected. When the enrichment culture time was 4 h, target bacteria with an initial contamination inoculation volume of 1.5 CFUs/mL could be detected in artificially contaminated samples. The method is fast and highly sensitive, and has not been applied to the detection of V. parahaemolyticus.

The rapid detection of pathogenic bacteria is important for the global public health field. Existing detection technologies are generally limited by log efficiency, high costs, and susceptibility to contamination. Advanced detection methods, such as surface plasmon resonance (SPR), have been proposed to break these limitations, with the advantages of fast detection speed and high sensitivity. SPR operates on the principle of attenuated total internal reflection at a metal–dielectric interface. This technique exploits the resonant absorption of incident photons by surface plasmons, facilitating the detection of tiny variations in the local refractive index by tracking the resonance condition change. This review provides a comprehensive overview of the development in the detection of foodborne pathogens using SPR technology, including a detailed discussion of the working principles of SPR, related instrumentation, and various detection methods. Specifically, this review focuses on a discussion of different SPR detection methods in terms of assembly processes, detection specificity, sensitivity, and detection limits in SPR biosensors, aiming to provide the readers with a fundamental knowledge of SPR.

Vibrio spp., known as typical marine pathogens, pose serious threats to human health worldwide due to their high incidence and prevalence. In this study, a quantitative detection method was developed for Vibrio parahaemolyticus, Vibrio vulnificus, Vibrio cholerae and Vibrio alginolyticus in aquatic products. A set of multiplex chamber-based digital PCR (cdPCR) primers and probes was designed based on new molecular targets of Vibrio species. The conditions of multiplex cdPCR were optimized and compared with those of multiplex qPCR. Under optimal conditions, the performance indicators, including the specificity, linear range, and detection limit, were systematically evaluated. The results indicate that the established method had 100% specificity among all tested strains and did not cross-react with other foodborne pathogens tested in parallel. The linear dynamic range was 10²–10⁶ CFU/mL, and the limits of detection (LODs) for V. parahaemolyticus, V. vulnificus, V. cholerae and V. alginolyticus were 5.40 × 10² CFU/mL, 3.05 × 10² CFU/mL, 4.55 × 10² CFU/mL and 1.32 × 10² CFU/mL, respectively. Furthermore, the feasibility of the proposed method was assessed in contaminated food samples, with a 10-fold improvement in detection sensitivity over that of the qPCR method under the conditions of this study. The entire detection process could be completed within 2 h. Taken together, the results of this study offer a readily adaptable diagnostic method, allowing for the simultaneous and quantitative detection of four pathogenic Vibrio species, which is promising for food safety and environmental monitoring.