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Lactic acid bacteria (LAB) are known to exhibit various beneficial roles in fermentation, serving as probiotics, and producing a plethora of valuable compounds including antimicrobial activity such as bacteriocin-like inhibitory substance (BLIS) that can be used as biopreservative to improve food safety and quality. However, the yield of BLIS is often limited, which poses a challenge to be commercially competitive with the current preservation practice. Therefore, the present work aimed to establish an optimised two-plasmid CRISPR/Cas9 system to redirect the carbon flux away from lactate towards compounds with antimicrobial activity by disrupting lactate dehydrogenase gene (ldh) on various strains of LAB. The lactic acid-deficient (ldhΔ) strains caused a metabolic shift resulting in increased inhibitory activity against selected foodborne pathogens up to 78 % than the wild-type (WT) strain. The most significant effect was depicted by Enterococcus faecalis-ldh∆ which displayed prominent bactericidal effects against all foodborne pathogens as compared to the WT that showed no antimicrobial activity. The present work provided a framework model for economically important LAB and other beneficial bacteria to synthesise and increase the yield of valuable food and industrial compounds. The present work reported for the first time that the metabolism of selected LAB can be manipulated by modifying ldh to attain metabolites with higher antimicrobial activity. • The development of a new mutant lactic acid bacteria (LAB) strains was attained by genetically disrupt lactate dehydrogenase (ldh) gene, to eliminate the production of lactic acid. • The deletion of the ldh was achieved by introducing an efficient two-plasmid CRISPR system. • The growth rate of the mutant LAB strains show significant changes due to the deletion of ldh. • The effects of metabolic alteration on Bacteriocin-Like Inhibitory Substance (BLIS) was significantly increase up to two-fold resulted from re-routing of the carbon flow towards BLIS pathway. [ABSTRACT FROM AUTHOR]

The list of standard abbreviations for JDS is available at adsa.org/jds-abbreviations-24. Nonstandard abbreviations are available in the Notes. Diarrheagenic Escherichia coli (DEC) is a kind of foodborne pathogen that poses a significant threat to both food safety and human health. To address the current challenges of high prevalence and difficult subtyping of DEC, this study developed a method that combined multiplex PCR with high-resolution melting (HRM) analysis for subtyping 5 kinds of DEC. The target genes are amplified by multiplex PCR in a single well, and HRM curve analysis was applied for distinct amplicons based on different melting temperature (Tm) values. The method enables discrimination of different DEC types based on characteristic peaks and distinct Tm values in the thermal melting curve. The assay exhibited 100% sensitivity and 100% specificity with a detection limit of 0.5 to 1 ng/μL. The results showed that different DNA concentrations did not influence the subtyping results, demonstrating this method owed high reliability and stability. In addition, the method was also used for the detection and subtyping of DEC in milk. This method streamlines operational procedures, shorts the detection time, and offers a novel tool for subtyping DEC. [ABSTRACT FROM AUTHOR]

Ribonucleases are in charge of the processing, degradation and quality control of all cellular transcripts, which makes them crucial factors in RNA regulation. This post-transcriptional regulation allows bacteria to promptly react to different stress conditions and growth phase transitions, and also to produce the required virulence factors in pathogenic bacteria. Campylobacter jejuni is the main responsible for human gastroenteritis in the world. In this foodborne pathogen, exoribonuclease PNPase (Cj PNP) is essential for low-temperature cell survival, affects the synthesis of proteins involved in virulence and has an important role in swimming, cell adhesion/invasion ability, and chick colonization. Here we report the crystallographic structure of Cj PNP, complemented with SAXS, which confirms the characteristic doughnut-shaped trimeric arrangement and evaluates domain arrangement and flexibility. Mutations in highly conserved residues were constructed to access their role in RNA degradation and polymerization. Surprisingly, we found two mutations that altered Cj PNP into a protein that is only capable of degrading RNA even in conditions that favour polymerization. These findings will be important to develop new strategies to combat C. jejuni infections. [Display omitted] • Cj PNP is a crucial enzyme for low temperature survival and chicken colonization. • We have determined the crystallographic structure of Cj PNP. • We have deciphered the mechanism of action of Cj PNP by mutational analysis. • We have found two Cj PNP mutants that are unable to perform RNA polymerization. • This findings can be crucial for developing strategies to combat C. jejuni infections. [ABSTRACT FROM AUTHOR]

This study examines how to obtain high levels of two major flavonoids, hesperidin and narirutin, from GY (green yuzu) powder and investigate their biological properties. We selected solvent extraction and enzyme treatment conditions before purification with an HP-20 Diaion column. We determined the appropriate extracted condition as 5 % GY with a 50 % ethanol solvent, and the enzyme treatment condition as 0.05 % RF enzyme for 24 h. Purification using 50 % ethanol as an eluent resulted in highly-enriched narirutin and hesperidin (50.78 % and 42.43 %, respectively). We also investigated the biological activities of extracts from each step, including the assessment of MMP-1 and tyrosinase inhibition activities, antibacterial properties, cytotoxicity, and anti-inflammatory activities. The extracts exhibited desirable properties such as skin elasticity, whitening effects, and anti-inflammatory effects in LPS-simulated RAW 264.7 cells. Highly purified extracts showed better antibacterial activity against food and dermatological pathogens. The results suggested that narirutin and hesperidin concentration and purity may influence the extracts' biological effects. Overall, this study provides valuable insights into the improved extraction and purification of narirutin and hesperidin from GY powder and their biological activities, highlighting their potential as a functional food and health product ingredients. [Display omitted] • Efficient extraction of hesperidin and narirutin from green yuzu was performed. • Enzyme treatment and Diaion HP-20 column chromatography provided 93 % yield. • Purified extracts showed antibacterial activity against food and skin pathogens. • Purified extract showed functions related to skin elasticity and whitening. • Anti-inflammatory activity of purified extract was observed in RAW264.7 cells. [ABSTRACT FROM AUTHOR]

Foodborne pathogen contamination is a main source of food safety issues. Rapid screening of pathogens in foods is imperative for preventing the outbreaks of foodborne diseases, which have posed serious threats to human health and economic development. This review aimed at recent progress of the microfluidic separators and biosensors for rapid and sensitive detection of foodborne pathogens. The key technologies for developing lab-on-chip separators and biosensors were illustrated. In particular, we systematically discussed the strategy of lab-on-chip magnetic separation for continuous-flow isolation of pathogenic bacteria. Challenges and future perspectives for practical applications of lab-on-chip technologies in foods were also discussed. Lab-on-chip devices are a promising tool to integrate separation, enrichment and detection in one single chip to facilitate "sample-in-answer-out" testing of pathogens in agri-food. [Display omitted] • Recent advances on microfluidic separators for isolation and enrichment of foodborne pathogens are summarized. • The strategy of microfluidic magnetic separation in continuous-flow isolation of pathogens are systematically discussed. • Recent progress of microfluidic electrochemical and optical biosensors for rapid detection of pathogens are summarized. • Challenges and future perspectives of practical application of lab-on-chip technology in agri-food are discussed. [ABSTRACT FROM AUTHOR]

Public health events caused by foodborne pathogens have brought substantial economic and social burdens. Conventional pathogen detection relies on a time-consuming culturing assay, laying back for the inspection requirements of the modern food industry. With portability, automation, high throughput, and cost-effectiveness, microfluidic systems provide a solution for advancing the diagnosis of foodborne pathogens. This review systematically summarizes the advances in microfluidic systems for detecting foodborne pathogens in the theme of substrates (i.e., glass/silicon, thermoplastics, hydrogels, paper, threads, the combination of various materials) enabled functions. We further discuss the extensions of microfluidic systems in the era of "big data" by integrating the Internet of Things (IoT) for in-time food safety inspection and warning. Additionally, we explore the future direction of microfluidic technology in detecting and monitoring foodborne pathogens. Towards the goal of applications, the fabrication of microfluidic devices has been gradually developed as low-cost, portable, and easy-to-use, from silicon/glass, and polymers, to hydrogel, paper, thread, and the combination of various materials. The materials with distinct physicochemical properties empower microfluidic systems with various advances for rapidly detecting foodborne pathogens. Microfluidic systems integrated with IoT have opened new opportunities for real-time food safety inspecting and warning. [Display omitted] • The functions of microfluidics empowered by fabrication material were summarized. • The advances in microfluidics for foodborne pathogen detection were reviewed. • The future of microfluidics in food safety surveillance was out looked. [ABSTRACT FROM AUTHOR]

Recently, food quality and safety have emerged as significant global issues. The rapid detection of food contaminants and spoilage markers has attracted increasing concerns. The grouping of illuminants based on the principle of aggregation-induced emission (AIE) mechanism for designing food safety and quality monitoring and analysis sensing platforms can address the shortcomings of conventional fluorophore aggregation-caused quenching (ACQ), offering the benefits of superb photoluminescence properties and trace-level detection accuracy. With the increasing demand for rapid and real-time monitoring in the food industry, such a robust sensing platform is becoming progressively important. In the current review, the luminescence mechanisms and structure design of AIE sensing platforms are first introduced. The sensing mechanisms are elucidated, including noncovalent interaction, solubility and viscosity change, specific recognition, and photophysical quenching/dequenching processes. Moreover, this review comprehensively summarizes the latest and representative applications of AIE sensing platforms in the detection of foodborne pathogens, environmental pollutants, biotoxins, agrochemicals, illegal additives, and spoilage markers. Lastly, the outlook and challenges of AIE sensing platforms are discussed. The emerging AIE sensing platform exhibits the outstanding advantages of rapidity, high sensitivity, and good accuracy in analytes detection, as well as predestined capabilities in constructing portable solid-state sensors. The combination of AIE sensing platforms with other technologies, such as immunosensing techniques, freshness labels, and smartphone platforms, can further facilitate the development of commercialized AIE sensing platforms, especially for portable, real-time, and on-site monitoring applications. From typical organic molecules to nature-derived AIE nanomaterials, the discovery, and synthesis of different types of AIE sensing substances are under active exploration to provide better solutions for food safety and quality inspection. • Review of the concept of AIE from the standpoint of sensing systems. • Introduction of AIE sensors in terms of structure design and sensing mechanism. • Summary of the novel trends of AIE sensors for food safety and quality detection. • Discussion on the future challenges in practical applications of AIE sensors. [ABSTRACT FROM AUTHOR]

The gram-positive bacterium Listeria monocytogenes is an important foodborne pathogen contaminating dairy products. Closely related to L. monocytogenes saprophytic Listeria spp. are also frequent contaminators of food and, particularly, dairy products. To distinguish L. monocytogenes from nonpathogenic Listeria spp. and other bacteria, a dot-immunoassay was developed. The immunoassay is based on the polyclonal antibody to the secreted form of the surface virulence-associated L. monocytogenes -specific InlB protein. To increase InlB production, bacteria were grown on the brain-heart infusion agar supplemented with 0.2% activated charcoal (BHIC agar). Direct plating of artificially contaminated raw milk samples on the BHIC agar followed by the dot-immunoassay allowed a rapid identification of L. monocytogenes in concentrations as little as 10 cfu/mL. Using the developed approach, preliminary results were obtained within 14 h, and the final results were obtained after 26 h. The dot-immunoassay was tested on L. monocytogenes strains belonging to different clonal complexes and phylogenetic lineages, Listeria spp., and other bacterial species. Results showed the exceptional specificity of the developed dot-immunoassay for the rapid identification of L. monocytogenes. [ABSTRACT FROM AUTHOR]

The advent of new technologies aimed at ensuring the food safety has increased in recent years. This trend has promoted the manufacturing of edible films and coatings with antibacterial agents. Among these agents, bacteriophages have been an attractive option to add into edible packaging and to preserve food safety. This review focuses on providing an overview on the manufacturing of edible film and coatings with antibacterial properties. Also focus on presents an appraisal of the studies that addressing the application of these bacteriophage-added edible packaging in food products and discussed the potential implications of these findings. Edible films and coatings with bacteriophages are suitable for the preservation of vegetables, fruits, meat, and cheese. These edible packaging enhance the food safety of these products by decreasing the growth of pathogenic bacteria. It was identified that the application of bacteriophages into edible films and coatings faces numerous challenges, including bacteriophage stability into the film, bacteriophage release from film to food, as well as the bacteriophage mobility and bacterial availability to promote the physical contact. [Display omitted] • Edible films and coatings with antibacterial properties are crucial for food safety. • Bacteriophages (phages) have been successfully applied in edible packaging. • Phages into films and coatings reduce the growth of foodborne pathogens. • Stability and release of phages are challenges to edible packaging application. • Food surface influences the phage activity. [ABSTRACT FROM AUTHOR]

Foodborne pathogens detection is important to ensure food safety and human health. In this study, we designed a comet structure to rapidly and sensitively detect foodborne Listeria monocytogenes. This method combined isothermal sequence exchange amplification (SEA) and surface-enhanced Raman spectroscopy. Listeria monocytogenes DNA could be rapidly amplified at a constant temperature via SEA with a pair of modified primers, which rendered the precise thermal control instrumentation unnecessary. Efficient SEA amplification generated a large number of DNA duplexes that could be easily captured by streptavidin-modified magnetic bead and AuMB@Ag-isothiocyanate fluorescein antibody (anti-FITC). AuMB@Ag-anti-FITC was used as a signal probe, which generated a significant excitation signal at 1,616 cm−1 for quantitative detection and analysis. The results displayed sensitive detection of L. monocytogenes in cheese from 2.0 × 101 cfu/mL to 2.0 × 106 cfu/mL within 1.0 h with a detection limit of 7.8 cfu/mL. Furthermore, this comet structure displayed the desirable specificity as its specific primers and amplified DNA ends were attached to streptavidin-modified magnetic beads and AuMB@Ag-anti-FITC, respectively. We expected that the method devised would provide a promising new approach to screening for L. monocytogenes and guarantee the microbiological safety of dairy products. [ABSTRACT FROM AUTHOR]

In this study, a fluorescent (FL) aptasensor was developed for on-site detection of live Salmonella typhimurium (S.T.) and Vibrio parahaemolyticus (V.P.). Complementary DNA (cDNA) of aptamer (Apt)-functionalized multicolor polyhedral oligomeric silsesquioxane-perovskite quantum dots (cDNA-POSS-PQDs) were used as encoded probes and combined with dual-stirring-bar-assisted signal amplification for pathogen quantification. In this system, bar 1 was labeled with the S.T. and V.P. Apts, and then bar 2 was functionalized with cDNA-POSS-PQDs. When S.T. and V.P. were introduced, pathogen-Apt complexes would form and be released into the supernatant from bar 1. Under agitation, the two complexes reached bar 2 and subsequently reacted with cDNA-POSS-PQDs, which were immobilized on MXene. Then, the encoded probes would be detached from bar 2 to generate FL signals in the supernatant. Notably, the pathogens can resume their free state and initiate next cycle. They swim between the two bars, and the FL signals can be gradually enhanced to maximum after several cycles. The FL signals from released encoded probes can be used to detect the analytes. In particular, live pathogens can be distinguished from dead ones by using an assay. The detection limits and linear range for S.T. and V.P. were 30 and 10 CFU/mL and 102–106 CFU/mL, respectively. Therefore, this assay has broad application potential for simultaneous on-site detection of various live pathogenic bacteria in water. [Display omitted] • A FL aptasensor was designed for simultaneous detection of live V.P. and S.T. • Multicolor PQD-encoded DNA probes were proposed for detection. • Dual-stirring-bar-assisted target recycling was used to improve sensitivity. • Superior specificity, anti-interference ability, and sensitivity were achieved. [ABSTRACT FROM AUTHOR]

The biogenic synthesis of silver nanoparticles (AgNPs) was performed using crude rosmarinic acid (RA) from plants as a reducing agent and coated with chitosan biopolymer. The prepared particles were characterized by ultraviolet–visible spectroscopy, Fourier transform infrared spectroscopy, X-ray diffraction (XRD), and transmission electron microscopy (TEM). A surface plasmon resonance peak at 430 nm indicates the emergence of AgNPs. XRD showed that the AgNPs were crystalline with an average crystalline size of 30 nm and TEM studies revelad that AgNPs were spherical without aggregation. The prepared CS-AgNPs exhibited good bactericidal properties against foodborne pathogens, such as Escherichia coli , Pseudomonas aeruginosa, and Vibrio parahaemolyticus. In particular, 100 μg/mL CS-AgNPs inhibited the growth of the selected bacteria and controlled their biofilm-forming ability. Band-aid cloth assay confirmed that the CS-AgNPs could be used in the medical field to prevent bacterial infections. The prepared CS-AgNPs increased the survival rate of Artemia species and exhibited antioxidant activity in conjunction with bactericidal properties against selected foodborne pathogens. • Chitosan capped AgNPs was biologically prepared and characterized. • Synthesized nanocomposite exhibited better antibacterial and antioxidant activity. • Synthesized nanocomposite exhibited antibiofilm activity. • Bio-toxicity of nanocomposite was evaluated. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus is a significant foodborne pathogen, commonly detected in milk and meat products. Conventional detection methods have limited sensitivity and specificity, which are time-consuming and susceptible to background interference from complex samples, and cannot effectively distinguish live and dead bacteria. Herein, we developed a novel adenosine triphosphate (ATP) bioluminescence method coupled with magnetic separation, which is based on phage-encoded endolysin LysSA163-CBD (CBD 163) for rapid and specific detection of viable Staphylococcus aureus. The expressed protein (CBD 163) was derived from the phage LSA2301 and was successfully expressed in Escherichia coli BL21 following an induction period of 4 h at 37 °C, with a molecular weight approximating 40 kDa. The optimal conditions for CBD-magnetic beads (cMBs) to capture S. aureus cells were determined to be 100 μL/mL cMBs at 25 °C for 30 min. The viable S. aureus cells were disrupted by the Cetyl trimethyl ammonium bromide (CTAB) to release intracellular ATP. Then, the ATP reacted with the firefly luciferase and D-Luciferin-based bioluminescence (BL) reagents solution to generate intensive BL signal. The CBD-magnetic separation-ATP bioluminescence (cMS-BL) assay was able to quickly detect viable S. aureus via ATP bioluminescence in 45 min, with a detection range from 5 × 103 to 5 × 107 CFU/mL and limit of detection (LOD) of 190 CFU/mL. Additionally, the cMS-BL method exhibited high specificity and anti-interference ability, which has been successfully applied to quantify S. aureus cells in crayfish-tail, chicken, and skim milk. These results demonstrate the potential of CBD 163 as a versatile and robust biorecognition element for rapid and specific detection of viable S. aureus in food matrices. The proposed phage-derived bacteria-binding proteins-based protocol for BL detection shows various advantages, including high sensitivity, simple operation, and the capability to distinguish live bacteria, providing a strategy for designing high-quality biorecognition element toward foodborne pathogens. [Display omitted] • Biorecognition element LysSA163-CBD (CBD 163) was efficiently expressed in E. coil BL21. • CBD 163 coupled with magnetic beads were synthesized to capture S. aureus cells specifically. • This CBD-magnetic separation-ATP bioluminescence assay can rapidly detect viable S. aureus in different food matrices. • This assay exhibited a wide linear range (5 × 103 to 5 × 107 CFU/mL) with a limit of detection of 190 CFU/mL. • This novel assay demonstrated sensitivity and specificity for the detection of viable S. aureus. [ABSTRACT FROM AUTHOR]

Gold nanoparticle-based lateral flow immunoassays (AuNP-LFIA) are widely used for pathogen monitoring to prevent foodborne illness outbreaks. However, conventional AuNP-LFIA exhibits poor sensitivity and limited quantitative capacity due to the low colorimetric signal intensity of AuNPs. Herein, we introduced a low-background gold in situ growth (GISG) strategy by lowering the pH of the growth solution to weaken the reducibility of hydroxylamine, thereby enhancing the sensitivity of AuNP-LFIA. Additionally, we developed a universal and manufacturable lateral flow device to streamline the GISG process. We applied this device to detect staphylococcal enterotoxin A (SEA), an exotoxin produced by Staphylococcus aureus. Under optimal conditions, the proposed device demonstrated superior practicality and excellent sensitivity for SEA detection, achieving a detection limit of 0.061 ng/mL with the total detection time of 37 min, showing 311 times more sensitive than the unamplified AuNP-LFIA. Furthermore, SEA detection in milk samples showed a strong correlation (R2 = 0.8845) with results obtained from a conventional ELISA kit. Therefore, this promising LFIA device offers a novel strategy with high sensitivity and practicality for in-field detection of Staphylococcus aureus and can be easily adapted for screening other foodborne pathogens. [Display omitted] • A low-background gold in situ growth strategy was developed to enhance the sensitivity of AuNP-LFIA. • A manufacturable lateral flow device was created to address the complexities associated with the gold in situ growth strategy. • The modified LFIA device was applied for the sensitive detection of SEA in milk. • The gold in situ growth-enhanced LFIA device offers approximately a 311-fold improvement in sensitivity compared to traditional AuNP-LFIA. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus contamination continues to be a harmful foodborne pathogen threatening of human health, and there is a growing need for rapid detection technologies. This study proposed a novel paper biosensor based on a polydiacetylene (PDA) polymer functionalized fibrinogen (Fg) for the detection of S. aureus in food sources. The fluorophore was developed based on the high binding ability of fibrinogen-binding proteins on the surface of S. aureus. This binding caused twisting in the PDA backbone, leading to changes in chromatic and fluorescent. The detection limit of this method was 50.1 CFU/mL for S. aureus- contaminated foodstuffs and 65.0 CFU/mL for the pure S. aureus culture, and the novelty came from its rapidity and selectivity for S. aureus compared to other foodborne bacteria. In summary, the present work provides a rapid detection method for S. aureus detection, which will help in addressing food safety-related issues. [Display omitted] • The colorimetric biosensor has proved its efficiency as a Staphylococcus aureus method. • The blue color change of PDA-Fg to red was associated with S. aureus concentrations. • The fluorescent emission of biosensors has appeared with the introduction of S. aureus into the sensing system. • The changes in the chemical groups PDA were monitored by FTIR spectroscopy. • PDA-Fg has proved its feasibility in detecting S. aureus in the contaminated samples. [ABSTRACT FROM AUTHOR]

[Display omitted] • A new method for detecting and differentiating Listeria monocytogenes serotypes on agar media. • NIR hyperspectral image data was used for classification model development. • Achieved over 80% accuracy for L. monocytogenes serotype classification. Among the severe foodborne illnesses, listeriosis resulting from the pathogen Listeria monocytogenes exhibits one of the highest fatality rates. This study investigated the application of near infrared hyperspectral imaging (NIR-HSI) for the classification of three L. monocytogenes serotypes namely serotype 4b, 1/2a and 1/2c. The bacteria were cultured on Brain Heart Infusion agar, and NIR hyperspectral images were captured in the spectral range 900–2500 nm. Different pre-processing methods were applied to the raw spectra and principal component analysis was used for data exploration. Classification was achieved with partial least squares discriminant analysis (PLS-DA). The PLS-DA results revealed classification accuracies exceeding 80 % for all the bacterial serotypes for both training and test set data. Based on validation data, sensitivity values for L. monocytogenes serotype 4b, 1/2a and 1/2c were 0.69, 0.80 and 0.98, respectively when using full wavelength data. The reduced wavelength model had sensitivity values of 0.65, 0.85 and 0.98 for serotype 4b, 1/2a and 1/2c, respectively. The most relevant bands for serotype discrimination were identified to be around 1490 nm and 1580–1690 nm based on both principal component loadings and variable importance in projection scores. The outcomes of this study demonstrate the feasibility of utilizing NIR-HSI for detecting and classifying L. monocytogenes serotypes on growth media. [ABSTRACT FROM AUTHOR]

Foodborne diseases are a global concern due to their high impact on morbidity, and mortality worldwide. The traditional method of bacterial detection is both time-consuming and labor-intensive. Alternative methods, such as immunoassays and polymerase chain reactions, have limited applicability due to the need for an enrichment process and expensive equipment. DNA-based biosensors have gained interest in the field of detection, due to their high rapidity, high sensitivity, and high specificity, which provides a promising technology for bacterial detection. To fabricate DNA-based biosensors, the immobilization of DNA probes on electrodes is a crucial process. This review delves into the three techniques of DNA probe immobilization, discussing their respective advantages and disadvantages. Furthermore, recent advancements in DNA-based biosensors are explored. The integration of advanced molecular techniques such as DNA nanostructures, DNA walkers, and the CRISPR-Cas system with DNA-based biosensors for foodborne pathogens is also discussed in this work. [Display omitted] • Review the recent advance in DNA-based foodborne bacterial detection. • Principle for immobilizing DNA probes and summarizing their benefits, and drawbacks. • Compare the latest diverse applications of DNA-based foodborne bacterial biosensors. • Advanced molecular techniques for foodborne pathogens are highlighted. [ABSTRACT FROM AUTHOR]

Salmonella is a globally prevalent foodborne pathogen, and adverse events caused by S. Enteritidis and S. Typhimurium are extremely common. With the emergence of drug resistance, there is an urgent need for efficient and specific lytic bacteriophages as alternative to antibiotics in clinical practice. In this study, phage P6 was isolated and screened from effluent and fecal samples from duck farm environments to specifically lyse the duck sources S. Typhimurium and S. Enteritidis. Phage P6 belongs to the genus Lederbergvirus, unclassified Lederbergvirus species. The phage P6 genome did not contained non-coding RNA, virulence genes and drug resistance genes, indicating that phage P6 was biologically safe for clinical applications. Phage P6 lysed 77.78% (28/36) of multidrug-resistant Salmonella and reduced biofilms formed by S. Enteritidis CVCC 3377, 4, and 24, and S. Typhimurium 44 by 44% to 75% within 3 h, and decreased Salmonella in duckling feces by up to 1.64 orders of magnitude. Prokaryotic expression of endolysin LysP6 lysed the chloroform-treated bacterial outer membrane from different serotypes of duck-derived Salmonella and E. coli standard strain ATCC 25922. The host range was expanded compared to phage P6, and the growth of Salmonella was effectively inhibited by LysP6 in conjunction with the membrane permeabilizer EDTA within 24 h. Therefore, phage P6 and phage-derived endolysins LysP6 are suitable for application as potent biocontrol agents to improve poultry health and food safety. [ABSTRACT FROM AUTHOR]

[Display omitted] The principle of detection through PCR-PfAgo system. • A detection method for Salmonella using PCR technology combined with PfAgo. • Select the hilA gene as a potential detection target for invasive Salmonella. • The PCR-PfAgo method meets fast detection, high sensitivity and low cost. Salmonella is a common zoonotic foodborne bacterial pathogen. In poultry farms, Salmonella can cause pullorum disease, which characterizes decreased appetite and diarrhea in chicks, accompanied by a high mortality rate in the first week of life itself. In pig farms, Salmonella mainly infects piglets and causes porcine paratyphoid, characterizing septicemia and necrotic enteritis and sometimes leading to severe symptoms such as encephalitis. This bacterium poses significant challenges to global public health and food safety. PfAgo (Pyrococcus furiosus Argonaute) is an Argonaute enzyme from the Pyrococcus furiosus , which is utilized in highly sensitive and accurate nucleic acid detection and analysis methods based on the polymerase chain reaction (PCR) combined with PfAgo. In this study, targeting the hilA gene, and can detect as low as 3 × 100 copies and 2.7 × 10°CFU (cells) with no cross-reactivity with other pathogens, it enhances the sensitivity of PCR. The PCR-PfAgo method was successfully applied to detect in clinical food samples including chicken and milk, and the results were consistent with those of traditional bacterial culture. The findings suggest that the PCR-PfAgo method has the potential to replace traditional detection methods and has broad application prospects in the detection of Salmonella pathogen. [ABSTRACT FROM AUTHOR]

The aim of this study was to characterize the pulp of Rheum ribes L. and to determine the effect of the pulp enriched with eugenol (1 %) or thymol (1 %) on the microbiological and physico-chemical quality of chicken breast fillets. Chicken breast fillets, inoculated with Listeria monocytogenes , Salmonella enterica subsp. enterica serovar Typhimurium, and Escherichia coli O157:H7 (~6.0 log 10), were marinated for 24 h in a mixture prepared from a combination of Rheum ribes L. pulp with eugenol or thymol. The quality parameters were analyzed for 15 days at +4 °C. The Rheum ribes L. pulp was found to have high antioxidant activity, high total phenolic content and contained 22 different phenolic substances, among which rutin ranked first. The pulp contained high levels of p-xylene and o-xylene as volatile substances and citric acid as an organic acid. The combination of Pulp + Eugenol + Thymol (PET) reduced the number of pathogens in chicken breast fillets by 2.03 to 3.50 log 10 on day 0 and by 2.25 to 4.21 log 10 on day 15, compared to the control group (P < 0.05). The marinating treatment significantly lowered the pH values of fillet samples on the first day of the study, compared to the control group (P < 0.05). During storage, TVB-N levels showed slower increase in the treatment groups compared to the control group (P < 0.05). In addition, the marinating process led to significant changes in physicochemical parameters such as water holding capacity, color, texture, cooking loss, and drip loss compared to the control group (P < 0.05). In conclusion, the results of this study showed that the pulp of Rheum ribes L., which has a high antioxidant capacity and contains various bioactive compounds. Furthermore, S. Typhimurium, E. coli O157:H7 and L. monocytogenes were inhibited considerably by marinating Rheum ribes L. pulp with a combination of eugenol and thymol. • The aim of this study was to evaluate the potential of rhubarb pulp as a marinade. • Rhubarb pulp contains four organic acids, 22 polyphenols and 35 volatiles. • Rhubarb pulp in combination with thymol and eugenol inhibited foodborne pathogens. • Marinating improved the microbiological and chemical status of the chicken breasts. [ABSTRACT FROM AUTHOR]

Foodborne illnesses caused by Salmonella and Staphylococcus aureus are a significant public health concern, leading to societal and economic repercussions. It is important to develop a simple and straightforward bacteria detection and identification method. A triple-probe multiplex rolling circle amplification technique has been developed to simultaneously detect Salmonella Typhimurium and S. aureus. This method utilizes fluorophore-labeled long padlock probes targeting S. Typhimurium invA and S. aureus glnA specific genes, along with a pH-based detection approach for direct visual identification. The multiplex hyperbranched saltatory rolling circle amplification assay at 30 °C has showed promising results with synthetic targets within 30 min and real bacteria within 2 h after establishing the detection settings. The assay is specific for S. aureus and S. Typhimurium, with a limit of detection of 39 μM for fluorescence and 78 μM for colorimetric. In the simulative test of this method for the detection of S. Typhimurium and S. aureus in milk, the limit of detection for the fluorescence signal after 2 h of amplification was 10 CFU/mL and 5 CFU/mL, respectively. The detection method was evaluated to be stable enough to detect pathogen for 3.29 months. Consequently, this triple-probe-multiplex rolling circle amplification method displays notable specificity, sensitivity, as well as ease of interpretation when testing food samples for harmful pathogens. [Display omitted] • Multiplex saltatory hyperbranched rolling circle low temperature amplification. • Biopolymer modified DNA extraction surface coupled with tunable hybrid signal. • One-step DNA extraction and exonuclease-free single tube RCA. • High speed, specificity, and sensitivity due to the triplex probe employed. • Enhanced in-field detection of Salmonella and Staphylococcus in milk. [ABSTRACT FROM AUTHOR]

For the past years, Salmonella has been one of the major foodborne pathogens in Europe, leading to the development of several control efforts to reduce its impact on human health. Poultry meat has been consistently implicated in foodborne cases of salmonellosis. One of the strategies to lessen the burden of salmonellosis in humans was the implementation of national control programs (NCPs) for Salmonella in broilers and turkeys aiming for reductions in these animal populations. In this paper, a description and comparison of the Salmonella surveillance and control programs that are currently implemented for the broiler and turkey chains in different European countries was performed. All the countries studied have set multiple surveillance and control actions for Salmonella at different stages of the broiler and turkey chains, namely the feed, farm and meat levels. Although most of the control programs are aligned with European Union (EU) regulations, some differences were observed, mostly regarding feed controls, farm surveillance schemes, target serovars and the handling of positive flocks. Overall, these differences had a regional pattern, with the Nordic countries having more detailed control programs with a zero-tolerance in meat. The remaining countries generally follow EU legislation, but in some cases, additional specifications were identified by this study. Despite the positive impact of these control programs on the reduction of human cases of salmonellosis, the decreasing tendency has reached a stall. The authors suggest that the NCPs are regularly revised within the framework of risk-based meat assurance systems, and the inclusion of additional target serovars which are simultaneously prevalent in broiler and turkey flocks and relevant in terms of public health within a country or a region. Furthermore, within the revision of NCPs, sampling schemes and strategies need to be consistent, following the risk management approach that has led to very low prevalences of Salmonella in poultry meat in some European countries. • Salmonella in poultry meat is one of the major sources of human infection. • All studied countries have surveillance and control programs for Salmonella in broilers and turkeys. • Nordic countries have more detailed programs, when compared to other European countries. • Salmonella National Control Programs should be revised regularly. [ABSTRACT FROM AUTHOR]

The application of dry aging on fish is increasing in restaurants worldwide. This process determines fluid loss, changes in muscle texture, and the development of a more concentrated flavour appreciated by consumers. Despite the growing popularity in the foodservice industry, there is a lack of data on potential risks and quality conditions of dry-aged fish. This study was aimed at assessing the safety for consumption of whole rainbow trout subjected to dry aging in a local restaurant (Turin, Italy). Quantitative microbiological analyses and detection of foodborne pathogens were performed on dorsal, ventral muscles, and skin samples at day 0, 3, 6, 10, and 14 of dry aging. Changes in water activity (a w), pH, colour, and biogenic amines in muscle were checked at each sampling point. Almost all the microbiological indicators were below the quantification limits in muscle until day 6 of dry aging, while higher but still acceptable loads were observed at day 10. At day 14, yeasts and moulds exceeded the acceptable level in one case (3.3 ± 0.4 log 10 CFU/g) in ventral muscle and counts of Pseudomonas spp. (from 6.1 ± 0.8 to 6.4 ± 0.5 log 10 CFU/g), total mesophilic (from 4.4 ± 0.3 to 5.3 ± 0.7 log 10 CFU/g), and psychrophilic (from 5.4 ± 0.1 to 5.8 ± 0.3 log 10 CFU/g) bacteria indicated the approaching of spoilage. Detection of Listeria monocytogenes and Staphylococcus aureus was always negative during the trials. Minimal changes of pH and colour were observed, while a w values followed a decreasing trend during dry aging. Putrescine was the only detectable amine, with a maximum concentration of 2.05 ± 0.02 mg/kg at day 14. In this study, dry-aged rainbow trout was safe for consumption and 10 days was the most appropriate curing time with 3 °C and 78% of relative humidity. Further studies at different environmental conditions and on other fish species are needed for a comprehensive risk assessment of this fast-growing practice in the restaurant sector. • Microbial loads remain within acceptable levels until day 10 of dry aging. • Listeria monocytogenes and Staphylococcus aureus are never detected during trials. • Putrescine is the only detectable amine, peaking 2.05 ± 0.02 mg/kg at day 14. • A ten-day curing period is the most suitable at 3 °C and 78% of relative humidity. [ABSTRACT FROM AUTHOR]

In this study, a colourimetric biosensor based on bacteriophage SapYZUM13 and an aminated Mn 3 O 4 (Mn 3 O 4 -NH 2) nanozyme was constructed and evaluated for its ability to detect Staphylococcus aureus in food. The biosensor had a detection time of 20 min, with a detection limit of 2 × 101 CFU/mL and recovery rate of 92.42–106.96%, indicating its high reliability and accuracy in detecting the food pathogen. Mechanistically, SapYZUM13@Mn 3 O 4 -NH 2 exhibited oxidase-mimicking capability, producing O 2 •– free radicals which oxidise 3,3′,5,5′-tetramethylbenzidine (TMB) to yield blue-coloured oxTMB. In the presence of S. aureus , the oxidase activity decreased remarkably owing to shielding of the nanozyme active sites. Moreover, SapYZUM13@Mn 3 O 4 -NH 2 could detect viable S. aureus from various sources, likely because of the special receptor-binding proteins of SapYZUM13 adsorbing to the wall teichoic acids on the S. aureus cell surface. Thus, SapYZUM13@Mn 3 O 4 -NH 2 has broad application prospects for the detection of viable S. aureus in various foods. [Display omitted] • SapYZUM13@Mn 3 O 4 -NH 2 can selectively, sensitively, and rapidly detect viable S. aureus. • SapYZUM13@Mn 3 O 4 -NH 2 exhibited accurate and reliable S. aureus counts in real food. • The detection mechanism of SapYZUM13@Mn 3 O 4 -NH 2 is due to the blockage of the active sites. • The recognition mechanism of SapYZUM13@Mn 3 O 4 -NH 2 is caused by the phage tail fibre. [ABSTRACT FROM AUTHOR]

[Display omitted] • Co-digestion of WAS and FW avoids acid inhibition with pH ranging from 6.4 to 8.7. • The highest CH 4 yield of 256.7 mL/g VS was achieved at WAS/FW=1. • The highest E. coli reduction of >6 log 10 CFU/mL is achieved at WAS/FW=1 and 2. • Low-pH induced free VFAs dominates E. coli inactivation in the acidogenesis stage. • Alkaline pH and the resultant NH 3 inactivate E. coli in the methanogenesis stage. Anaerobic co-digestion of waste activated sludge (WAS) with food waste (FW) represents a viable and effective approach to energy recovery and pollutant elimination. Little is known about the effect of alternating change of acid/alkaline conditions on pathogens inactivation within the co-digestion system of WAS and FW. In this study, pH changes during batch anaerobic co-digestion with WAS/FW ratios varying from 0.4 to 8, and their impact on pathogen inactivation were investigated using Escherichia coli as a model pathogen. At WAS/FW=1 and 2, serious acidification (pH<5.5) was effectively mitigated, and the pH transitioned from 6.4 during the acidogenesis stage to >8.5 during the methanogenesis stage. WAS/FW=1 achieved the highest methane yield (256.7 mL/g VS), which was 38.9 % greater than that in their mono-digestion scenario. A 2.37 − 3.87 log 10 reduction in E. coli abundance occurred in all tested conditions during the acidogenesis stage, and a further reduction to an undetectable level was achieved only in WAS/FW reaching 1 subsequently. Low pH induced free volatile fatty acids dominated E. coli inactivation during the acidogenesis stage. During the methanogenesis stage, the combined action of OH− and the resultant free ammonia contributed to E. coli inactivation with free ammonia being more efficient in its action. This research contributes to the advancement of sustainable and secure management of organic wastes. [ABSTRACT FROM AUTHOR]

Rapid and accurate detection of pathogenic Salmonella in food and environmental sources is of paramount importance to protect public health from salmonellosis. However, most current assays are labor-intensive, time-consuming, and require expensive instruments and experienced operators. In this work, we developed a PicoArray-based colony-forming unit (CFU) quantification system for rapid and ultrasensitive Salmonella detection. To enable on-site detection, we employed syringe-vacuum-assisted sample loading and oil-induced sample digitization to stochastically confine single bacteria in the picoliter-level wells of the PicoArray device (containing 40,233 wells). Such a single bacterial confinement enables the isolation and characterization of individual bacteria from a large population. In addition, the confinement of bacteria in small volumes allows rapid accumulation of metabolites, thereby eliminating the pre-incubation step and reducing the time required to detect bacteria. By combining it with a selective medium for the isolation of Salmonella and a fluorescent redox indicator (e.g. , resazurin) to facilitate the detection of individual microcolonies, the PicoArray device can rapidly and precisely detect Salmonella. As a proof-of-concept, we demonstrated the ability of our platform for rapid, sensitive, and specific detection of Salmonella in spiked milk samples. Our platform offers the possibility of close monitoring of pathogens in food or drinks at the consumer end because it allows a non-trained operator to detect foodborne pathogens in the clinically relevant range in the home setting. [Display omitted] • A portable digital colony forming unit (CFU) quantification platform has been developed. • A resazurin-amplified detection accelerates the quantification of microcolonies in picowells. • The platform coupled with selective media enables precise quantification of specific bacteria. • Our digital CFU platform allows on-site, sensitive detection of foodborne pathogens. [ABSTRACT FROM AUTHOR]

Rising living standards have heightened the demand for fresh, safe, and nutritious food, necessitating advanced food quality and safety monitoring technologies. Electrospun nanofibers (ENFs) are gaining increasing attention as key materials in the development of high-performance sensing platforms for food safety and quality monitoring owing to their high surface area, porosity, and customizable structures. This review provides a comprehensive overview of recent advances in the application of ENFs for food analysis. It highlights their roles and mechanisms in detecting environmental toxins, identifying foodborne pathogens, analyzing food components, and monitoring freshness. Furthermore, the benefits, limitations, and future development directions of ENFs in food analysis are discussed. ENFs can serve as effective materials for sample pretreatment, enhancing detection sensitivity, streamlining procedures, and reducing detection times through target analyte adsorption and enrichment. They also hold considerable potential as scaffolds for sensors that enable rapid and accurate detection of various food analytes. However, challenges related to standardization and environmental impact must be addressed to transition ENFs from laboratory research to practical applications. [Display omitted] • Applications of ENFs in food analysis are comprehensively reviewed. • Roles and merits of ENFs in food analysis are summarized. • The latest developments in detecting toxins, pathogens, and food freshness using ENFs are reviewed. • Remaining limitations and potential of ENFs for food analysis are discussed. [ABSTRACT FROM AUTHOR]

Antimicrobial agents are effective weapons against foodborne pathogens; however, antimicrobial resistance (AMR) is increasing globally. The bacterial cell envelope is a key antimicrobial target, and understanding its structure and dynamics is crucial for detecting AMR pathogens and promoting rational antimicrobial design. Advanced microscopic techniques, such as cryo-electron microscopy (cryo-EM) and atomic force microscopy (AFM), provide unique real-time insights into cell envelope dynamics under antimicrobial stress, setting a benchmark for combating pathogens and AMR. This review presents an overview of conventional and advanced microscopic techniques for bacterial cell envelope analysis, focusing, for each technique, the prospects, limitations, and current improvements. Conventional microscopic techniques, such as scanning electron microscopy, transmission electron microscopy, fluorescence microscopy, and confocal laser scanning microscopy, are predominant in foodborne bacterial analysis, mainly for morphological and ultrastructural observation. Advanced microscopic techniques, such as cryo-EM and AFM, are scarcely exploited owing to their expensive instrumentation, technical expertise requirements, and complex data processing. Nevertheless, cryo-EM reveals novel AMR mechanisms of numerous multidrug efflux pumps and resolves key membrane structures in foodborne pathogens. AFM captures antimicrobial action in bacterial membranes, providing insights into the structure‒activity relationship of membrane-targeting antimicrobial agents. Additionally, cryo-EM and AFM can be integrated with artificial intelligence and machine learning tools, improving overall performance, reducing the need for technical expertise, and simplifying data processing. Therefore, advanced microscopic techniques are essential for developing novel strategies to overcome the threats of foodborne pathogens and AMR. • Advances in microscopic techniques for foodborne bacterial analysis reviewed. • Advanced microscopy reveals foodborne pathogen responses and resistance to antimicrobials. • Cryo-EM and AFM unravel the molecular antimicrobial mechanisms under physiological conditions. • Advanced microscopy is critical to combatting foodborne pathogens and antimicrobial resistance. [ABSTRACT FROM AUTHOR]

[Display omitted] • The nanozymes are categorized from the perspective of nanomaterials. • Colorimetric biosensors are presented using the nanozyme's catalytic activity. • The recent developments of nanozymes in colorimetric biosensing of various pathogenic bacteria are discussed. • The challenges and future prospects of the nanozymes-based colorimetric biosensors are deliberated. The prompt identification of foodborne pathogens is essential for safeguarding food safety and reducing financial repercussions. Nanozymes, characterized as nanomaterials exhibiting enzyme-like activity, are increasingly emerging as viable alternatives to natural enzymes as a result of their benefits like flexible design, regulated catalytic activity, good biocompatibility and high stability. The remarkable catalytic function of nanozymes has led to their extensive utilization in the development of colorimetric biosensors. Colorimetric sensors offer numerous advantages, including naked-eye visibility, ease of operation, portability, and high sensitivity. They have emerged as a potent and efficient tool for rapid detection, enabling the development of innovative colorimetric sensing strategies through the integration of nanozymes with colorimetric sensors. In this review, the recent advancements in nanozyme research and its utilization in colorimetric biosensing of pathogenic bacteria are analyzed. Firstly, nanozymes are presented in a systematic classification based on various nanomaterials, and their characteristics are elaborated upon in detail. Following this, the study centers on examining the construction approaches of nanozymes exhibiting peroxidase (POD), oxidase (OXD), superoxide dismutase (SOD), and catalase (CAT) activities for colorimetric biosensing of diverse pathogenic bacteria. Finally, the present challenges and future prospects concerning the enhanced utilization of nanozymes in colorimetric biosensors are deliberated in the last section. [ABSTRACT FROM AUTHOR]

[Display omitted] • PDA BIP-based biosensor for sensitive and specific detection of L.monocytogenes. • BUHNPs-modified GCE amplifies electrochemical biosensor signals. • Synergistic interaction of aptamer with BIP film shortens target recognition time. • The LOD of the electrochemical biosensor was 1.0 CFU·mL−1. • The method has been applied to the detection of L.monocytogenes in food samples. Food safety problems caused by foodborne pathogens pose a serious threat to human health and its sensitive detection is an urgent issue. Herein, a polydopamine film-based electrochemical biosensor was constructed for ultrasensitive detection of Listeria monocytogenes (L. monocytogenes). Bimetallic silver-gold sea urchin-like hollow nanoparticles (BUHNPs) were first synthesized and loaded on the surface of a glassy carbon electrode, and then aptamers (apt) were modified on the BUHNPs, followed by depositing of dopamine to form polydopamine of bacterial imprinted polymers (PDA BIPs) film, and finally three-dimensional molecularly imprinted films with cavities were achieved after template elution. The presence of L. monocytogenes resulted in a change in the current signal, indicating that the bacteria had entered the cavity. The biosensor was able to detect L.monocytogenes at 101-106 CFU·mL−1 with a detection limit of 1.0 CFU·mL−1. The sensor can achieve rapid detection within 7 min, and the synergistic effect of BIP and aptamer enhances the recognition of the target, showing a great potential for the detection of L. monocytogenes in real food samples. [ABSTRACT FROM AUTHOR]

Salmonella enterica serovar Enteritidis (S. Enteritidis) poses a significant threat as a prevalent foodborne pathogen globally. Timely diagnostic methods are essential for effective management of S. Enteritidis infections. Herein, we establish a one-step LAMP-CRISPR/Cas12b method for a rapid, accurate, and convenient detection of S. Enteritidis. The entire reaction requires only 45 min without the need for specialized equipment. The platform exhibited 100% specificity with no cross-reactivity observed among 19 other Salmonella serovars and 9 non- Salmonella bacterial pathogens. Our method achieved a limit of detection of 30 copies per reaction. Compared with the traditional culture methods in practical analysis. S. Enteritidis positivity rates identified by one-step LAMP-CRISPR/Cas12b platform were 2.88% (6/208) including 3.14% (4/127) in chicken, 4.44% (2/45) in duck, and 0.0% (0/36) in pork, and a 100% concordance was observed between the traditional culture method and one-step LAMP-CRISPR/Cas12b platform. In summary, our findings underscore the simplicity and accuracy of the one-step LAMP-CRISPR/Cas12b platform for S. Enteritidis detection, with potential applications in resource-limited settings. • The simplicity and accuracy of the one-step LAMP-CRISPR/Cas12b platform for S. Enteritidis detection. • The one-step LAMP-CRISPR/Cas12b platform presents 100% consistency with traditional culture method. • A limit of detection of 30 copies per reaction mixture and can be finished within 1h. • The platform exhibited 100% specificity (3/3) with no cross-reactivity observed among other bacterial pathogens. [ABSTRACT FROM AUTHOR]

Glyco sylated bacterio cins , known as glycocins, were first discovered in 2011. These bioactive peptides are produced by bacteria to gain survival advantages. They exhibit diverse types of glycans and demonstrate varied antimicrobial activity. Currently, there are 13 experimentally known glycocins, with over 250 identified in silico across different bacterial phyla. Notably, glycocins are recognized for their glycan-mediated antimicrobial activity, proving effective against drug-resistant and foodborne pathogens. Many glycocins contain rare S-linked glycans. Glycosyltransferases (GTs), responsible for transferring sugar to glycocins and involved in glycocin biosynthesis, often cluster together in the producer's genome. This clustering makes them valuable for custom glycoengineering with diverse substrate specificities. Heterologous expression of glycocins has paved the way for the establishment of microbial factories for glycopeptide and glycoconjugate production across various industries. In this review, we emphasize the primary roles of fully and partially characterized glycocins and their glycosylating enzymes. Additionally, we explore how specific glycan structures facilitate these functions in antibacterial activities. Furthermore, we discuss newer approaches and increasing efforts aimed at exploiting bacterial glycobiology for the development of food preservatives and as replacements or complements to traditional antibiotics, particularly in the face of antibiotic-resistant pathogenic bacteria. • Analysis of recent developments in glycocin research and applications, based on papers and patents over the last decade. • Current methods for enhancing glycocin synthesis and purification and glycoengineering. • Future prospective and applications of glycocins in various sectors. [ABSTRACT FROM AUTHOR]

[Display omitted] • Discover the latest trends in rapid detection through a review of recent breakthroughs in identifying foodborne pathogens. • The innovation and limitations of different detection methods and their relationship were summarized. • CRISPR/Cas applications in rapid detection are summarized, showing innovation in food safety. • Molecular biology techniques offer high specificity, sensitivity, and rapidity for improved foodborne pathogens detection. In recent years, foodborne diseases have posed a serious threat to human health, and rapid detection of foodborne pathogens is particularly crucial for the prevention and control of such diseases. This article offers a detailed overview of the development of detection techniques for foodborne pathogens, transitioning from traditional microbiological culture methods to the current array of techniques, including immunological, molecular biological, and biosensor-based methods. It summarizes the technical principles, advantages, disadvantages, and research progress of these diverse methods. Furthermore, the article demonstrates that the combination of different methods enhances the efficiency and accuracy of pathogens detection. Specifically, the article focuses on the application and advantages of combining CRISPR/Cas systems with other detection methods in the detection of foodborne pathogens. CRISPR/Cas systems, with their high specificity, sensitivity, and ease of operation, show great potential in the field of foodborne pathogens detection. When integrated with other detection techniques such as immunological detection techniques, molecular biology detection techniques, and biosensors, the accuracy and efficiency of detection can be further improved. By fully utilizing these tools, early detection and control of foodborne diseases can be achieved, enhancing public health and preventing disease outbreaks. This article serves as a valuable reference for exploring more convenient, accurate, and sensitive field detection methods for foodborne pathogens, promoting the application of rapid detection techniques, and ensuring food safety and human health. [ABSTRACT FROM AUTHOR]

The prevention of foodborne pathogens has been a significant aspect of food safety. Among these pathogens, Salmonella stands out as one of the most representative strains, and the prevention of its contamination is paramount for protecting food safety. Here, we successfully developed an electrochemical immunosensor based on the synergistic amplification of internal and external dual signals for the rapid detection of Salmonella in food. The Fe 3 O 4 @Prussian blue (PB) core-shell nanomaterial, synthesized via the in-situ generation of PB on the Fe 3 O 4 surface through acid etching, exhibited good stability and redox activity, enabling its integration with [Fe(CN) 6 ]3−/4− ionic tags for dual-signal synergistic amplification of current signals. The microstructure and elemental composition of the Fe 3 O 4 @PB core-shell nanomaterial were analyzed using relevant instruments. In optimal conditions, this strategy demonstrated a broad detection range from 7.375 × 101 to 7.375 × 107 CFU mL −1, with a low detection limit of 9.912 CFU mL −1. Furthermore, the sensor we developed shows good stability, excellent selectivity, and reproducibility, and has great application potential in the detection of foodborne pathogens. [Display omitted] • Dual-signal synergistic strategy remarkably improves the sensitivity of the sensor. • Immunosensor has wide linear range from 7.375 × 101 to 7.375 × 107 CFU mL −1. • The detection limit of Salmonella is 9.912 CFU mL −1. • This study provides some ideas for the detection of other pathogens. [ABSTRACT FROM AUTHOR]

Vibrio alginolyticus (V. alginolyticus) is a common foodborne pathogen that poses a serious threat to food safety, therefore it urgently needs to be effectively controlled. In this study, a novel peptides-zinc nanocomposite (P@ZnNP) with antibacterial activity was fabricated from crimson sapper scales peptides (CSSP). The effect of P@ZnNP on V. alginolyticus was expected from the antibacterial and anti-virulence properties, as well as corresponding mechanism. The minimum inhibitory concentration (MIC) and time-killing curve data showed that P@ZnNP with a concentration of 0.23 mg/mL exhibited significant antibacterial activity against V. alginolyticus. The analysis of the antibacterial mechanism of planktonic V. alginolyticus illustrated that it acted through membrane damage. The virulence related results revealed that the motility, biofilm formation, extracellular polysaccharide production, and alkaline serine protease synthesis of V. alginolyticus were significantly inhibited. Furthermore, in fresh shrimp models, P@ZnNP could reduce bacterial abundance and attenuate virulence. Given these findings, it is believed that P@ZnNP has the potential to become an antibacterial agent against V. alginolyticus in the food industry. • Peptides-zinc nanocomposite (P@ZnNP) was fabricated by Zn–N and Zn–O covalent bonds. • P@ZnNP exerted efficacy against V. alginolyticus through membrane damage mechanism. • Reducing bacterial motility, biofilm formation, EPS and virulence production. • Inhibiting V. alginolyticus on shrimp models significantly. • P@ZnNP may be useful as excellent antibacterial agent for control V. alginolyticus. [ABSTRACT FROM AUTHOR]

Proper preservation of nutrient-rich raw food from microbial contamination is a major challenge faced by the food industry. The use of antibiotics and chemicals to address the issue of contamination in fresh produce and meat has led to antibiotic-resistant pathogens. In the current study, the anti-biofilm effect of bio-oil aqueous phase (BOAP) collected during pyrolysis of seaweed was evaluated against some major food-borne pathogens for its biofilm inhibition and eradication potential in mono- and dual-species biofilms. The results showed that BOAP could eradicate almost 75% of dual-species biofilms. Further, the preservative effect of the BOAP was examined using appropriate food samples (pineapple and prawns). There was profound reduction in the bacterial load in the food samples which shows that BOAP has effective food preservation property. SEM analysis also revealed the drastic reduction in biofilm in BOAP treated food samples. In future, natural preservatives and BOAP can be combined to increase shelf-life of the above food samples. • Bio-oil Aqueous Phase (BOAP) obtained from pyrolysis of seaweed biomass. • BOAP eradicated biofilms formed by foodborne pathogens. • Effective reduction of bacterial load in fruit and sea-food models. • Potential for enhancing the shelf life of fresh cut pineapple and prawns. [ABSTRACT FROM AUTHOR]

Foodborne pathogens have been a serious threat to human health and safety for a long time. However, the complex food substrate undoubtedly increases the difficulty of bacterial detection at low concentrations. How to effectively improve the sensitivity of bacterial detection is a major problem that needs to be solved. In this study, the aptamer of emetic Bacillus cereus (B. cereus) was used as the recognition molecule to construct a magnetic capture probe. Utilizing the strong affinity between target bacteria and the aptamer, a fluorescence sensor based on aptamer-rolling circle amplification (Apt-RCA) was established for the detection of emetic B. cereus in milk spiked samples. In this study, DNA-functionalized magnetic beads were first synthesized. The strong affinity between the aptamer and the target bacteria makes it to be used not only as an antibody substitute for capturing the target bacteria, but also to compete the complementary DNA (cDNA) strand of the aptamer on the DNA-functionalized magnetic bead as a primer for the subsequent RCA reaction. RCA reaction can produce RCA products rich in a large number of G-quadruplex sequences. The fluorescent dye thioflavin T (ThT) can bind to the G-quadruplex structure and produce an obvious fluorescence signal. The fluorescence signal amplification strategy based on the magnetic separation platform and RCA reaction successfully realized the sensitive detection of emetic B. cereus in actual sample. Under optimal conditions, the limits of detection (LODs) of pure bacterial solution and milk samples were 1.4 × 102 CFU/mL and 2.2 × 103 CFU/mL, respectively. This method has the advantages of short detection time, simple operation and good specificity, which provides a new way of thinking for the detection of foodborne pathogens. [Display omitted] • Aptamer functionalized magnetic beads were synthesized as magnetic capture probes. • Magnetic capture probes combined with RCA were used to construct fluorescent aptasensor. • Aptamer complementary strands can be used as primers to initiate RCA reactions. [ABSTRACT FROM AUTHOR]

The rapid and accurate identification of live pathogens with high proliferative ability is in great demand to mitigate foodborne infection outbreaks. Herein, we have developed an ultrasensitive image-based aptasensing array to directly detect live Salmonella typhimurium (S.T) cells. This method relies on the long-range orientation of surfactant-decorated liquid crystals (LCs) and the superiority of aptamers (apt ST). The self-assembling of hydrophobic surfactant tails leads to a perpendicular/vertical ordered film at the aqueous/LC interface and signal-off response. The addition of apt ST perturbed LCs' ordering into a planar/tilted state at the aqueous phase due to electrostatic interactions between the surfactant with the apt ST , and a signal-on response. Following the conformational switch of apt ST in the presence of live S. typhimurium , a relative reversing signal-off response was observed upon the target concentration. This aptasensor could promptly confirm the presence of S. typhimurium without intricate DNA-extraction or pre-enrichment stats over a linear range of 1–1.1 × 106 CFU/mL and a detection limit of 1.2 CFU/mL within ∼30 min. These results were successfully validated using molecular and culture-based methods in spiked-milk samples, with a 92.61–104.61 % recovery value. Meanwhile, the flexibility of this portable sensing platform allows for its development and adoption for the precise detection of various pathogens in food and the environment. [Display omitted] • An innovative portable LC-aqueous aptasensor is developed for viable bacteria detection. • A signal-off mode was introduced into optical aptasensors based on aptamer-target interaction. • A calibration curve from 100 −106 CFU/mL was utilized for S. typhimurium with a 1.1 CFU/mL LOD. • The aptasensor performs the sensing without any sample pre-enrichment steps. • The LC-aqueous aptasensor achieved sensitive and superbright rapid (∼30 min) detection of live S. typhimurium in food. [ABSTRACT FROM AUTHOR]

High-resolution and efficient typing for Laribacter hongkongensis (L. hongkongensis) is essential for epidemiological investigation of such emerging foodborne pathogens. Clustered regularly interspaced short palindromic repeats (CRISPR) typing is an innovative molecular method that shows great promise for L. hongkongensis typing. Here, we explored the CRISPR typing method by combining CRISPR1 and CRISPR2 loci to characterize a collection of 109 L. hongkongensis isolates from humans and animals and compared it to current molecular methods such as pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). The results showed that all three methods have high discriminatory power (diversity index was 0.9902 for PFGE, 0.9663 for CRISPR and 0.9562 for MLST); strong congruence was observed between them (Rand index was 0.969 between CRISPR and PFGE, 0.953 between CRISPR and MLST, 0.958 between PFGE and MLST). CRISPR typing could well distinguish the isolates in the same STs or PFGE profiles, and the genetic information contained by the CRISPR array is useful for deep phylogenetic typing. We demonstrate that rapid CRISPR typing is a practical genetic fingerprinting tool with high resolution, comparable ease of use and lower cost, ability to track the source of various groups of L. hongkongensis strains and indication of genetic characteristics. • CRISPR typing is congruent with PFGE and MLST with great discriminating power. • All the three approaches are suitable for L. hongkongensis transmission analyses. • CRISPR typing is faster, easier of use and lower cost than other two approaches. [ABSTRACT FROM AUTHOR]

Foodborne illnesses, caused by harmful microorganisms in food, are a significant global health issue. Current methods for identifying these pathogens are both labor-intensive and time-consuming. In this research, we devised a swift and precise detection technique using recombinase polymerase amplification combined with a lateral flow dipstick (RPA-LFD) for three foodborne pathogens found in meat. By employing a dedicated detection device, RPA-LFD allows for the rapid analysis of DNA from Escherichia coli O157 (E. coli O157), Salmonella , and Shigella —pathogens that are prohibited in food. The detection thresholds for E. coli O157, Salmonella , and Shigella are 0.168 fg/μl (1.04 CFU/ml), 0.72 fg/μl (27.49 CFU/ml), and 1.25 fg/μl (48.84 CFU/ml), respectively. This method provides a short detection window, operates at low temperatures, follows simple procedures, and exhibits high sensitivity. Our study establishes the RPA-LFD method for simultaneously identifying the nucleic acid of three foodborne pathogens, offering an efficient solution for quickly identifying multiple contaminants. • A comprehensive nucleic acid detection box, based on the RPA-LFD principle. • The detection box is characterized by its disposable and lid-sealed nature. • Simultaneous detection of three pathogenic bacteria in food • Rapid nucleic acid detection of Salmonella, Shigella, and E. coli O157 • Demonstrated a short detection time, high sensitivity, and ease of observation [ABSTRACT FROM AUTHOR]

Developing innovative films and coatings is paramount for extending the shelf life of numerous food products and augmenting the barrier and antimicrobial properties of food packaging materials. Many synthetic chemicals used in active packaging and food storage have the potential to leach into food, posing long-term health risks. It is imperative for active packaging materials to inherently possess biological protective properties to ensure food quality and safety throughout its storage. Bacteriophages, or simply phages, are bacteria-eating viruses that serve as promising natural biocontrol agents and antimicrobial bioadditives in food packaging materials, specifically targeting bacterial foodborne pathogens. These phages are generally recognized as safe (GRAS) by regulatory authorities for food safety applications. They exhibit targeted action against various Gram-positive and -negative foodborne pathogens, including Bacillus spp., Campylobacter spp., Escherichia coli , Listeria monocytogenes , Salmonella spp., Shigella spp., and Vibrio spp., associated with foodborne spoilage and illness without affecting the beneficial microbes. Phage cocktails can be applied directly on food surfaces, incorporated into food packaging materials, or utilized during food processing treatments. Unlike chemical agents, phage activity increases proportionally with the rise in pathogenic bacterial populations. Researchers are exploring various packaging materials to deliver phages with broad host range, stability, and viability ensuring their effectiveness in safeguarding various food systems. The effectiveness of phage immobilization or encapsulation on active food packaging materials depends on various factors, including the characteristics of polymers, the choice of solvents, the type of phage, and its loading efficiency. Factors such as the orientation of phage immobilization on substrates, pH, temperature, exposure to carbohydrates and amino acids, exopolysaccharides, lipopolysaccharides, and metals can also influence phage activity. In this review, we comprehensively discuss the various active packaging systems utilizing bacteriophages as natural biocontrols and antimicrobial bioadditives to reduce the incidence of foodborne illness and enhance consumer confidence in the safety of food products. [ABSTRACT FROM AUTHOR]

[Display omitted] • Detection of Yersinia enterocolitica in bivalve mollusks by a reference test (ISO 10273:2017). • Validation "in house" of the molecular method employed to detect pathogenic Yersinia enterocolitica in bivalve mollusks. • Detection of Yersinia enterocolitica in real samples of bivalve mollusks. Yersinia enterocolitica (Ye) is a foodborne pathogen isolated from humans, food, animals, and the environment. Yersiniosis is the third most frequently reported foodborne zoonosis in the European Union. Ye species are divided into six biotypes 1A, 1B, 2, 3, 4, and 5, based on biochemical reactions and about 70 serotypes. Biotype 1A is non-pathogenic, 1B is highly pathogenic, and biotypes 2–5 have moderate or low pathogenicity. The reference analysis method for detecting pathogenic Ye species underestimates the presence of the pathogen due to similarities between Yersinia enterocolitica -like species and other Yersiniaceae and/or Enterobacteriaceae , low concentrations of distribution pathogenic strains and the heterogeneity of Yersinia enterocolitica species. In this study, the real-time PCR method ISO/TS 18867 to identify pathogenic biovars of Ye in bivalve molluscs was validated. The sensitivity, specificity and accuracy of the molecular method were evaluated using molluscs experimentally contaminated. The results fully agree with those obtained with the ISO 10273 method. Finally, we evaluated the presence of Ye in seventy commercial samples of bivalve molluscs collected in the Gulf of Naples using ISO/TS 18867. Only one sample tested resulted positive for the ail gene, which is considered the target gene for detection of pathogenic Ye according to ISO/TS 18867. Additionally, the presence of the ystB gene, used as target for Ye biotype 1A, was assessed in all samples using a real-time PCR SYBR Green platform. The results showed amplification ystB gene aim two samples. [ABSTRACT FROM AUTHOR]

[Display omitted] • Survival of L. monocytogenes in minced pork to single or sequential environmental stresses after cold stress was explored. • Thermal and gastric fluid resistance of L. monocytogenes was evaluated after sequential stress. • NaCl adaptation reduced the resistance of L. monocytogenes to LA and Nisin stress, whereas LA induced the opposite trend. • Sequential pressure reduces the effect of a single pressure on the thermal resistance of L. monocytogenes. • Sequential stress of LA 1.25-NaCl 50 renders thermal-treated L. monocytogenes insensitive to simulated gastric fluid. In this study, Listeria monocytogenes from minced pork was evaluated for changes in resistance to thermal treatment and gastric fluid following environmental stresses during food processing. Bacteria were exposed to cold stress, followed by successive exposures to different stressors (lactic acid (LA), NaCl, or Nisin), followed by thermal treatments, and finally, their gastrointestinal tolerance was determined. Adaptation to NaCl stress reduced the tolerance of L. monocytogenes to subsequent LA and Nisin stress. Adaptation to LA stress increased bacterial survival in NaCl and Nisin-stressed environments. Bacteria adapted to Nisin stress showed no change in tolerance to subsequent stress conditions. In addition, treatment with NaCl and LA enhanced the thermal tolerance of L. monocytogenes , but treatment with Nisin decreased the thermal tolerance of the bacteria. Almost all of the sequential stresses reduced the effect of a single stress on bacterial thermal tolerance. The addition of LA and Nisin as a second step of stress reduced the tolerance of L. monocytogenes to gastric fluid, whereas the addition of NaCl enhanced its tolerance. The results of this study are expected to inform processing conditions and sequences for meat preservation and processing and reduce uncertainty in risk assessment of foodborne pathogens due to stress adaptation. [ABSTRACT FROM AUTHOR]

[Display omitted] • Biofilm and planktonic L. monocytogenes are sensitive to rhamnolipids. • pH and NaCl influence antimicrobial action of rhamnolipids. • Rhamnolipids are more effective at pH 5.0. • NaCl enhances rhamnolipid action mainly at pH above 6.0. • Rhamnolipid self-assembly is associated with antimicrobial activity. The use of natural and sustainable additives, that are less aggressive to the environment, is a trend in the food industry. Rhamnolipids (RL) biosurfactants have shown potential for controlling food pathogens however, due to the presence of free carboxyl groups, the pH and ionic strength may influence the properties of such surfactants. In this study, we describe the antimicrobial activity of RL under different pH values and NaCl concentrations, towards both planktonic and biofilms of Listeria monocytogenes. RL were effective at pH 5.0 and the addition of 5 % NaCl improved the bactericidal efficacy for planktonic and sessile cells. The effect of NaCl was more pronounced at pH above 6 showing a significant increase in RL antimicrobial activity. At pH 7.0 planktonic population was eradicated by RL only when salt was present whereas biofilm viability was decreased by 5 log with MBIC varying from > 2500.0 mg/L (RL) to 39.0 mg/L (RL + 5 % NaCl). Larger vesicular and lamellar RL self-assembly structures were predominant when NaCl was present, suggesting their association with the antimicrobial activity observed. The pH and ionic strength of the medium are important parameters to be considered for the development of RL-based strategies to control L. monocytogenes. [ABSTRACT FROM AUTHOR]

Multiplex, sensitive, and rapid detection of pathogens is crucial for ensuring food safety and safeguarding human health, however, it remains a significant challenge. This study proposes a c oncanavalin A -assisted m ultiplex d igital a mplification (CAMDA) assay for simultaneous quantitative detection of multiple foodborne bacteria. The CAMDA assay enables the simultaneous detection of six foodborne pathogens within 1.1 h and the limit of detection is 101 CFU/mL. Furthermore, the CAMDA assay exhibits high specificity, with a rate of 97 % for Bacillus cereus and 100 % for other pathogens tested in this study. Moreover, practical application validation using eight milk powder samples demonstrates that the accuracy of the CAMDA assay reaches 100 % when compared to qPCR results. Therefore, our developed CAMDA assay holds great potential for accurate and rapid detection of multiple pathogens in complex food matrices while also promoting the utilization of microfluidic chips in food investigation. [Display omitted] • A self-priming multiplex digital PCR chip was developed. • Canavalin A-based multiplex digital amplification (CAMDA) assay was established. • It can accurately detect six foodborne pathogens simultaneously. • The detection time is within 1.1 h and the limit of detection is 101 CFU/mL. [ABSTRACT FROM AUTHOR]

Salmonellosis is one of the most common foodborne diseases worldwide, with the ability to infect humans and animals. Antimicrobial resistance (AMR) and, particularly, multidrug resistance (MDR) among Salmonella enterica poses a risk to human health. Antimicrobial use (AMU) regulations in livestock have been implemented to reduce AMR and MDR in foodborne pathogens. In this study, we used an integrated machine learning approach to investigate Salmonella AMR and MDR patterns before and after the implementation of AMU restrictions in agriculture in the United States. For this purpose, Salmonella isolates from cattle in the National Antimicrobial Resistance Monitoring System (NARMS) dataset were analysed using three descriptive models consisting of hierarchical clustering, network analysis, and association rule mining. The analysis showed the impact of the United States' 2012 extra-label cephalosporin regulations on AMR trends and revealed a distinctive MDR pattern in the Dublin serotype. The results also indicated that each descriptive model provides insights on a specific aspect of resistance patterns and, therefore, combining these approaches make it possible to gain a deeper understanding of AMR. [ABSTRACT FROM AUTHOR]

• For the first time, we devised a Thermus thermophilus Argonaute (Tt Ago)-powered centrifugal microfluidic chip biosensor for sample-in-answer-out detection of foodborne bacterial pathogens, termed ASAP. • Tt Ago was multifunctional and was tasked to be nucleic acid signal transductor and amplifier in ASAP. • The limit of detection (LOD) of ASAP was down to 1 CFU/mL, and the turnaround time was 17 min. • The one-pot fashion of ASAP largely avoided carry-over and cross-contamination; • The multiple-channel feature of ASAP permitted simultaneous multiplexed detection. Foodborne pathogens endanger public health and rapid, sensitive and accurate detection of them is vital. Argonaute stands in the frontline as the next generation nucleic acid detection tool, bearing a few comparable advantages. Centrifugal microfluidic chips (CMCs) use centrifugal force to achieve liquid flowing, mixing and reaction, eliminating complicated designs of valves and pumps. In this study, for the first time, we devised a Thermus thermophilus A rgonaute (Tt Ago)-powered centrifugal microfluidic chip reaction biosensor for S ample-in- A nswer-out detection of P athogen S. aureus (termed as ASAP) with ultrahigh rapidity and sensitivity in a one-pot fashion. The samples subjected to testing were mixed with a home-made nucleic acid fast extraction reagent and then the mixture was injected into the sample cell of CMC, which was centrifuged down into the reaction cell. The reaction cell was preloaded with both LAMP (Loop-mediated isothermal amplification) and Tt Ago systems. As such, the species-specific nuc genes were amplified by LAMP, while the Tt Ago performed site-specific cleavage to output fluorescent (FL) signals. The sample-to-answer time was 17 min, and the limit of detection (LOD) reached 1 CFU/mL. ASAP was able to perform simultaneous multiplexed detection and up to 16 samples can be detected at the same time. ASAP reduced reagent consumption and minimized the influence of carry-over and cross-contamination. ASAP was capable of detecting S. aureus in foods, but also detecting physiological samples from infect, holding great promise for practical applications. Overall, our work has enriched the Argonaute-powered biosensing and CMC biosensor technology by providing a conceptually novel bacterial detection platform. [ABSTRACT FROM AUTHOR]

The potential advantages of using 3-class sampling plans in the management of pathogens in food has been discussed for years. Using a 3-class plan, some lots would be accepted if, among a total number of samples n , the pathogen was detected in no more than a predefined number of samples c (such as c = 1) exceeding a microbiological limit m , and no sample was at a level above a predefined maximal concentration M. We built upon and extended beyond published methodologies and developed a web application to evaluate the performance of 2-class and 3-class sampling plans. We showed that there is little benefit of a 3-class plan over a 2-class plan with equal values for m , n and c under the classical assumption of a lognormal distribution of bacteria concentration in the lot with a standard deviation of 0.8 log cfu/g, for m value set to "absence in 25g" and M set to values > 1 cfu/g. In this situation, the lot will almost always be rejected based on the criterion "> c detected samples" and will almost never (or seldom) be rejected based on only the criterion "at least one sample > M cfu/g". However, the benefit of a 3-class plan is realized when the distribution of pathogens is not a simple lognormal but includes a proportion of samples contaminated beyond the lognormal distribution, at levels > M. When this is the case, the performance of a 3-class plan relative to the 2-class plan with equal values of m , n , and c improves, then becomes essentially equivalent, as the proportion of highly contaminated samples increases. The behavior of 3-class plan may be useful in such situations. While such distributions of pathogens have been observed across lots in the food supply, better characterization of within-lot contamination distribution in food is needed to better evaluate the performance of sampling plans. • Interest in 3-class sampling plans for testing pathogens in food is long standing. • There is little benefit of a 3-class plan under classical assumptions. • 3-class plans are beneficial in preferential rejection of highly contaminated lots. • More data on bacteria concentration distribution in foodstuffs are needed. • We provide a web application to show how 2-class and 3-class sampling plans differ. [ABSTRACT FROM AUTHOR]

Concern about food safety triggers demand on rapid, accurate and on-site detection of foodborne pathogens. Among various fluorescent probes for detection, carbon dots (CDs) prepared by carbonization of carbon-rich raw materials show extraordinary performance for their excellent and tailorable photoluminescence property, as well as their facilely gained specificity by surface customization and modification. CDs-based fluorescent probes play a crucial role in many pathogenic bacteria sensing systems. In addition, microfluidic technology with characteristics of portability and functional integration is expected to combine with CDs-based fluorescent probes for point-of-care testing (POCT), which can further enhance the detection property of CDs-based fluorescent probes. Here, this paper reviews CDs-based bacterial detection methods and systems, including the structural modulation of fluorescent probes and pathogenic bacteria detection mechanisms, and describes the potential of combining CDs with microfluidic technology, providing reference for the development of novel rapid detection technology for pathogenic bacteria in food. • Applications of CDs in detection of foodborne pathogen are systematically reviewed. • Fabrication and post-modification of CDs to construct fluorescent sensors are included. • Mechanisms of recognition and response towards pathogenic bacteria are summarized. • Potential combination of CDs with microfluidic systems is elaborated. [ABSTRACT FROM AUTHOR]

The microbial water quality of poultry water supplies can be correlated to bird health, livability, overall performance, and human food safety. On-farm microbial evaluations were conducted to understand microbial hygiene of poultry waterlines based on water sanitation practices. With on-farm study I, 2 farms were selected: Farm A and Farm B; that did not practice water sanitation consistently during the flock grow-out period. Prevalence tests for specific pathogens- Salmonella, E coli, Campylobacter, Listeria, and Staphylococcus were performed for both farms by collecting swab samples post bird harvest from waterlines and plating in specific growth media. With the on-farm study II, 1 broiler farm with 4 barn units that treated water (Chlorine [ Cl ]= ∼ 1 ppm) during the flock grow-out period and flushed waterlines between flocks using concentrated chlorine solution (>1,000 ppm) was selected. Swab samples (1 from each barn, n = 4; each sampling occasion) were collected on 3 occasions-before flushing, after flushing, and at the end of the grow-out period (d 42) to understand biofilm growth nature in poultry waterlines. Additionally, a separate in-vitro study was conducted to understand the efficacy of a commonly used poultry drinking water sanitizer product (sodium hypochlorite, 8.25%) against specific foodborne pathogens : Salmonella Enteritidis (SE), E. coli and Listeria. Results from the on-farm study I showed that Farm A and Farm B were positive for Listeria and Staphylococcus, whereas these farms were found negative for other species tested. The findings from on-farm study II showed that biofilm reestablished (>4 log 10 cfu/mL) in waterlines by the end of grow-out cycle despite waterline cleaning and consistent water sanitation during flock grow-out period. The in-vitro test showed that the efficacy of chlorine-based water sanitizer at the field application dose rates was affected by pathogen load in water and pathogen types. The overall results from on-farm or in-vitro studies indicated that poultry growers need to emphasize consistent and robust poultry drinking water sanitation practice in their farms to keep the acceptable bacterial levels in poultry water supplies or water systems. [ABSTRACT FROM AUTHOR]