Origin of fecal contamination in lettuce and strawberries: From microbial indicators, molecular markers, and H. pylori.

Around 2 billion people utilize a water source contaminated with fecal-origin microorganisms, used for both human consumption and irrigation of crops. In Colombia, the water from the Bogotá River is employed for irrigating agricultural products, including raw-consumption foods like strawberries and lettuce. This poses a risk to the end consumer, as these foods are marketed as fresh products ready for direct consumption without undergoing any disinfection or cooking treatment. The aim of this study was to determine the origin of fecal contamination in strawberries and lettuce irrigated with surface waters from Cundinamarca, Colombia, using non-human and human molecular markers, along with Helicobacter pylori (H. pylori). A total of 50 samples were collected, 25 of strawberries and 25 of lettuce, taken from crops, markets, and supermarkets. Microbiological indicators (bacterial and viral) were detected through cultivation techniques, and Microbial Source Tracking (MST) markers and H. pylori were detected through PCR. The results of our study demonstrate the presence of Escherichia coli (E. coli) (12.5 %), Enterococcus (≥25 %), spores and vegetative forms of Spores of sulphite-reducing Clostridia (SRC) (≥37.5 %), coliphages (≥12.5 %), and Salmonella sp. (≥12.5 %), in both strawberries and lettuce. In the different samples analyzed, molecular markers were detected to differentiate the source of fecal contamination above 12.5 % (HF187, CF128, ADO and DEN) and H. pylori between 0 % and 25 %, highlighting deficiencies in the production chain. of food, and the risks they pose to food security. Highlighting deficiencies in the food production chain and the risks they pose to food safety.

Association analysis of antibiotic and disinfectant resistome in human and foodborne E. coli in Beijing, China.

The widespread use of chemical disinfectants and antibiotics poses a major threat to food safety and human health. However, the mechanisms of co-transmission of antimicrobial resistance genes (ARGs) and biocides and metal resistance genes (BMRGs) of foodborne pathogens in the food chain is still unclear. This study isolated 343 E. coli strains from animal-derived foods in Beijing and incorporated online data of human-derived E. coli strains from NCBI. Our results demonstrated a relatively uniform distribution of strains from various regions in Beijing, indicating a lack of region-specific clustering. Additionally, predominant sequence types varied between food- and human-derived strains, suggesting a preference for different hosts and environments. Phenotypic association analysis showed that the chlorine disinfectants peroxides had a significant positive correlation with tetracyclines. Many more ARGs and BMRGs were enriched in human-associated E. coli compared with those in chicken- and pork-origin. The quaternary ammonium compounds (QACs) resistance gene qacEΔ1 had a strong correlation with aminoglycoside resistance gene aadA5, folate pathway antagonist resistance gene dfrA17, sul1 and macrolide resistance gene mph(A). The correlation results indicated a significant association between the copper resistance gene cluster pco and the silver resistance gene cluster sil. Coexistence of many resistance genes was observed within the qacEΔ1 gene structure, with qacEΔ1-sul1 being the most common combination. Our findings demonstrated that the epidemiological spread of resistance is affected by a combination of heavy metals, disinfectants and antibiotic use, suggesting that the prevention and control strategies of antimicrobial resistance need to be multifaceted and comprehensive.

Prevalence and characterization of aminoglycoside resistance gene aph(2")-If-carrying Campylobacter jejuni.

Campylobacter jejuni is recognized as a significant foodborne pathogen, and recent studies have indicated a rising trend of aminoglycosides resistance gene aph(2″)-If among C. jejuni isolates from food-producing animals in China. However, systematic information about aph(2″)-If-positive C. jejuni from food-producing animals and other sources worldwide based on whole-genome analysis remains a knowledge gap. In this study, we aimed to analyze the worldwide distribution, genetic environment and phylogenetic tree of aph(2″)-If by utilizing Whole Genome Sequencing (WGS) data obtained, coupled with information in the GenBank database. A total of 160C. jejuni isolates in the GenBank database and 14C. jejuni isolates in our laboratory carrying aph(2″)-If gene were performed for further analysis. WGS analysis revealed the global distribution of aph(2″)-If among C. jejuni from 6 countries. Multilocus Sequence Typing(MLST) results indicated that 70 STs were involved in the dissemination of aph(2″)-If, with ST10086 being the predominant ST. Whole-genome Multilocus Sequence Typing(wg-MLST) analysis according to times, countries, and origins of C. jejuni isolation further demonstrated a close relationship between aph(2″)-If carrying C. jejuni isolates from farm and food. The findings also revealed the existence of 32 distinct types of genetic environments surrounding aph(2″)-If among these isolates. Notably, Type 30, characterized by the arrangement ISsag10-deoD-ant(9)-hp-hp-aph(2″)-If, emerged as the predominant genetic environment. In conclusion, our analysis provides the inaugural perspective on the worldwide distribution of aph(2″)-If. This resistance gene demonstrates horizontal transferability and regional diffusion in a clonal pattern. The close association observed among aph(2″)-If-positive C. jejuni strains isolated from poultry, food, and clinical environments underscores the potential for zoonotic transmission from these isolates.

Improvement of quantitative microbiological risk assessment (QMRA) methodology through integration with gaenetic data.

Quantitative microbiological risk assessment (QMRA) methodology aims to estimate and describe the transmission of pathogenic microorganisms from animals and food to humans. In microbiological literature, the availability of whole genome sequencing (WGS) data is rapidly increasing, and incorporating this data into QMRA has the potential to enhance the reliability of risk estimates. This study provides insight into which are the key pathogen properties for incorporating WGS data to enhance risk estimation, through examination of example risk assessments for important foodborne pathogens: Listeria monocytogenes (Lm), Salmonella, Campylobacter and Shiga toxin-producing Escherichia coli. By investigating the relationship between phenotypic pathogen properties and genetic traits, a better understanding was gained regarding their impact on risk assessment. Virulence of Lm was identified as a promising property for associating different symptoms observed in humans with specific genotypes. Data from a genome-wide association study were used to correlate lineages, serotypes, sequence types, clonal complexes and the presence or absence of virulence genes of each strain with patient's symptoms. We also investigated the effect of incorporating WGS data into a QMRA model including relevant genomic traits of Lm, focusing on the dose-response phase of the risk assessment model, as described with the case/exposure ratio. The results highlighted that WGS studies which include phenotypic information must be encouraged, so as to enhance the accuracy of QMRA models. This study also underscores the importance of executing more risk assessments that consider the ongoing advancements in OMICS technologies, thus allowing for a closer investigation of different bacterial subtypes relevant to human health.

Antimicrobial Efficacy of Allium cepa and Zingiber officinale Against the Milk-Borne Pathogen Listeria monocytogenes.

Listeria monocytogenes is an important food-borne pathogen that causes listeriosis and has a high case fatality rate despite its low incidence. Medicinal plants and their secondary metabolites have been identified as potential antibacterial substances, serving as replacements for synthetic chemical compounds. The present studies emphasize two significant medicinal plants, Allium cepa and Zingiber officinale, and their efficacy against L. monocytogenes. Firstly, a bacterial isolate was obtained from milk and identified through morphology and biochemical reactions. The species of the isolate were further confirmed through 16S rRNA analysis. Furthermore, polar solvents such as methanol and ethanol were used for the extraction of secondary metabolites from A. cepa and Z. officinale. Crude phytochemical components were identified using phytochemical tests, FTIR, and GC-MS. Moreover, the antibacterial activity of the crude extract and its various concentrations were tested against L. monocytogenes. Among all, A. cepa in methanolic extracts showed significant inhibitory activity. Since, the A. cepa for methanolic crude extract was used to perform autography to assess its bactericidal activity. Subsequently, molecular docking was performed to determine the specific compound inhibition. The docking results revealed that four compounds displayed strong binding affinity with the virulence factor Listeriolysin-O of L. monocytogenes. Based on the above results, it can be concluded that the medicinal plant A. cepa has potential antibacterial effects against L. monocytogenes, particularly targeting its virulence.

Detection of resistance and virulence plasmids in Campylobacter coli and Campylobacter jejuni isolated from North Carolina food animal production, 2018-2019.

Campylobacter remains the leading cause of bacterial foodborne illness in the U.S. and worldwide. Campylobacter plasmids may play a significant role in antimicrobial resistance (AMR) and virulence factor distribution, and potentially drive rapid adaptation. C. coli (n = 345) and C. jejuni (n = 199) isolates collected from live cattle, swine, turkey, and chickens, poultry carcasses at production, and retail meat in N.C. were analyzed to determine plasmid prevalence, extrachromosomal virulence and AMR genes, and the phylogeny of assembled plasmids. Putative plasmids ranging from <2 kb to 237kb were identified with virulence factors present in 66.1% (228/345) C. coli and 88.4% (176/199) C. jejuni plasmids (promoting adherence, invasion, exotoxin production, immune modulation, chemotaxis, mobility, and the type IV secretion system). AMR genes were identified in 21.2% (73/345) C. coli and 28.1% C. jejuni plasmids (conferring resistance to tetracyclines, aminoglycosides, beta-lactams, nucleosides, and lincosamides). Megaplasmids (>100 kb) were present in 25.7% (140/544) of the isolates and carried genes previously recognized to be involved with interspecies recombination. Our study highlights the extensive distribution and diversity of Campylobacter plasmids in food animal production and their role in the dissemination of biomedically important genes. Characterizing Campylobacter plasmids within the food animal production niche is important to understanding the epidemiology of potential emerging strains.

Antimicrobial effect of dried koruk (Vitis vinifera L.) pomace against food-borne pathogens inoculated in kofte.

This study aimed to investigate the antimicrobial effects of various concentrations of dried koruk pomace (1%, 1.5%, and 2%) used in kofte formulations. To detect the inactivation effect of dried koruk pomace on food-borne pathogens, kofte samples were separately inoculated with Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella Typhimurium at high and low levels. During the storage period, E. coli O157:H7, L. monocytogenes, and S. Typhimurium counts of the samples inoculated with high levels were decreased in the range of 1.1-1.3 log CFU/g, 0.1-0.2 log CFU/g and 1.1-1.2 log CFU/g, respectively. When low inoculum levels were used, E. coli O157:H7 and S. Typhimurium counts of the samples were reduced to an undetectable level at the beginning and after 2 months of storage, respectively. Although L. monocytogenes counts of the samples were reduced to an undetectable level immediately, the counts were increased during storage till the end of the fifth month. The results indicated that dried koruk pomace was more effective in eliminating E. coli O157:H7 and S. Typhimurium than L. monocytogenes inoculated in kofte. The results suggest that using dried koruk pomace in kofte formulations is an effective method to improve the safety of the products, especially when low levels of contamination occur.

Nanotechnology in Food and Plant Science: Challenges and Future Prospects.

Globally, food safety and security are receiving a lot of attention to ensure a steady supply of nutrient-rich and safe food. Nanotechnology is used in a wide range of technical processes, including the development of new materials and the enhancement of food safety and security. Nanomaterials are used to improve the protective effects of food and help detect microbial contamination, hazardous chemicals, and pesticides. Nanosensors are used to detect pathogens and allergens in food. Food processing is enhanced further by nanocapsulation, which allows for the delivery of bioactive compounds, increases food bioavailability, and extends food shelf life. Various forms of nanomaterials have been developed to improve food safety and enhance agricultural productivity, including nanometals, nanorods, nanofilms, nanotubes, nanofibers, nanolayers, and nanosheets. Such materials are used for developing nanofertilizers, nanopesticides, and nanomaterials to induce plant growth, genome modification, and transgene expression in plants. Nanomaterials have antimicrobial properties, promote plants' innate immunity, and act as delivery agents for active ingredients. Nanocomposites offer good acid-resistance capabilities, effective recyclability, significant thermostability, and enhanced storage stability. Nanomaterials have been extensively used for the targeted delivery and release of genes and proteins into plant cells. In this review article, we discuss the role of nanotechnology in food safety and security. Furthermore, we include a partial literature survey on the use of nanotechnology in food packaging, food safety, food preservation using smart nanocarriers, the detection of food-borne pathogens and allergens using nanosensors, and crop growth and yield improvement; however, extensive research on nanotechnology is warranted.

Evaluation of Antibacterial Activity of Selenium Nanoparticles against Food-Borne Pathogens.

Selenium is an essential micronutrient for all mammals and plays an important role in maintaining human physiological functions. Selenium nanoparticles (SeNPs) have been shown to demonstrate antioxidant and antimicrobial activity. The objective of this study was to explore whether SeNPs have the potential to be used as food preservatives with which to reduce food spoilage. SeNPs were synthesized through ascorbic acid reduction of sodium selenite (Na2SeO3) in the presence of bovine serum albumin (BSA) as a capping and stabilizing agent. The chemically synthesized SeNPs had a spherical conformation with an average diameter of 22.8 ± 4.7 nm. FTIR analysis confirmed that the nanoparticles were covered with BSA. We further tested the antibacterial activity of these SeNPs against ten common food-borne bacteria. A colony-forming unit assay showed that SeNPs exhibited inhibition on the growth of Listeria Monocytogens (ATCC15313) and Staphylococcus epidermidis (ATCC 700583) starting at 0.5 µg/mL, but higher concentrations were required to slow down the growth of Staphylococcus aureus (ATCC12600), Vibrio alginolyticus (ATCC 33787), and Salmonella enterica (ATCC19585). No inhibition was observed on the growth of the other five test bacteria in our study. Our data suggested that the chemically synthesized SeNPs were able to inhibit the growth of some food-borne bacteria. The size and shape of SeNPs, method of synthesis, and combination of SeNPs with other food preservatives should be considered when SeNPs are to be used for the prevention of bacteria-mediated food spoilage.

Emergence of lnu(C) variant conferring lincomycin resistance in Campylobacter coli of chicken origin.

Lincomycin is widely used in respiratory and gastrointestinal infection in veterinary medicine and food animal production. Campylobacter members are vital foodborne pathogens causing campylobacteriosis, and the resistance to lincosamides is seldom reported. To date, only the rRNA methyltransferase Erm(B) has been confirmed to be associated with lincosamides resistance in Campylobacter. In this study, we identified a lnu(C) variant conferring lincomycin resistance in this pathogen of chicken origin. The Lnu(C) encoded by this gene variant showed substitution at position 8 (Asn8Lys), 11 (Phe11Leu) and 112 (Leu112Phe), when compared with the firstly reported Lnu(C) from Streptococcus agalactiae. Cloning of the lnu(C) variant into lincosamide-susceptible Campylobacter jejuni NCTC 11168 confirmed its function in conferring resistance to lincomycin with the 32-fold increased MICs. Sequencing analysis showed that the lnu(C) variant was located within a MTnSag1-like transposon together with insLNU, which is inserted between panB and cj0299 genes on the chromosome. lnu(C) gene was distributed among C. coli globally, and various STs were involved in the dissemination of lnu(C). Although transposition mediated by MTnSag1-like transposon failed to occur, the horizontal transfer mediated by natural transformation and reservoir for resistance genes may facilitate their adaptation to the antimicrobial selection pressure in chickens, which should not be ignored.

Serotype-dependent adhesion of Shiga toxin-producing Escherichia coli to bovine milk fat globule membrane proteins.

Shiga toxin-producing Escherichia coli (STEC) are food-borne pathogens that can cause severe symptoms for humans. Raw milk products are often incriminated as vehicule for human STEC infection. However, raw milk naturally contains molecules, such as the milk fat globule membrane and associated proteins, that could inhibit pathogen adhesion by acting as mimetic ligands. This study aimed to: (i) evaluate the capability of STEC cells to adhere to bovine milk fat globule membrane proteins (MFGMPs), (ii) highlight STEC surface proteins associated with adhesion and (iii) evaluate the variation between different STEC serotypes. We evaluated the physicochemical interactions between STEC and milk fat globules (MFGs) by analyzing hydrophobic properties and measuring the ζ-potential. We used a plate adhesion assay to assess adhesion between MFGMPs and 15 Escherichia coli strains belonging to three key serotypes (O157:H7, O26:H11, and O103:H2). A relative quantitative proteomic approach was conducted by mass spectrometry to identify STEC surface proteins that may be involved in STEC-MFG adhesion. The majority of E. coli strains showed a hydrophilic profile. The ζ-potential values were between -3.7 and - 2.9 mV for the strains and between -12.2 ± 0.14 mV for MFGs. Our results suggest that non-specific interactions are not strongly involved in STEC-MFG association and that molecular bonds could form between STEC and MFGs. Plate adhesion assays showed a weak adhesion of O157:H7 E. coli strains to MFGMPs. In contrast, O26:H11 and O103:H2 serotypes attached more to MFGMPs. Relative quantitative proteomic analysis showed that the O26:H11 str. 21,765 differentially expressed five outer membrane-associated proteins or lipoproteins compared with the O157:H7 str. EDL933. This analysis also found strain-specific differentially expressed proteins, including four O26:H11 str. 21,765-specific proteins/lipoproteins and eight O103:H2 str. PMK5-specific proteins. For the first time, we demonstrated STEC adhesion to MFGMPs and discovered a serotype effect. Several outer membrane proteins-OmpC and homologous proteins, intimin, Type 1 Fimbriae, and AIDA-I-that may be involved in STEC-MFG adhesion were highlighted. More research on STEC's ability to adhere to MFGMs in diverse biological environments, such as raw milk cheeses and the human gastrointestinal tract, is needed to confirm the anti-adhesion properties of the STEC-MFG complex.

Phenotypic and genotypic antibiotic resistance patterns in Salmonella Typhimurium and its monophasic variant from pigs in southern Spain.

The high incidence of human salmonellosis and multi-drug resistant (MDR) strains of Salmonella Typhimurium (ST) is of concern to global public and animal health. Our research, by means of the broth microdilution method, evaluated the Minimum Inhibitory Concentration (MIC) distribution of 12 antimicrobials against a collection of 73 ST and mST and S. typhimurium monophasic variant 4,[5],12:i:- (mST) isolates from slaughtered pigs reared in extensive systems in southern Spain, and also 12 resistance-associated genes or antimicrobial resistance (AMR) determinants using qPCR. Our data revealed that 98.6% of strains were MDR, with resistance to cephalothin/tetracycline/sulfamethoxazole-trimethoprim/ampicillin/chloramphenicol being the most common pattern (55.6%). Regarding AMR determinants, the most significantly (p < 0.05) genes detected by qPCR were sul1 and aadA2 (89% of strains positive), aadA1 and dfrA12 (87.7%), and blaTEM and tet(B) (86.3% and 84.9%, respectively). Up to date information on ST antimicrobial resistance patterns is essential for epidemiological surveillance programs to support animal and public health. The high number of MDR isolates and variability regarding resistance determinants revealed in this study highlights the role of animals reared in extensive systems as a source of resistant Salmonella strains.

Characterization of plasmid-mediated quinolone resistance genes and extended-spectrum beta-lactamases in non-typhoidal Salmonella enterica isolated from broiler chickens.

Background and Aim: Antibiotic-resistant Salmonella is a public health concern. Fluoroquinolones and extended-spectrum beta-lactams are widely used for the treatment of Salmonella infections. This study focused on the detection of plasmid-mediated quinolone resistance (PMQR) and extended-spectrum beta-lactamase (ESBL) genes among multidrug-resistant (MDR) Salmonella enterica isolated from broilers. Materials and Methods: A total of 40 non-typhoidal S. enterica isolates were collected from 28 broiler chicken farms in four Iraqi Governorates. These isolates were examined for their susceptibility to 10 antimicrobial agents by disk-diffusion method followed by polymerase chain reaction assay for the detection of PMQR determinants and ESBLs genes. Results: Salmonella strains revealed high levels of resistance to the following antibiotics: Nalidixic acid 100%, levofloxacin (LEV) 97.5%, amoxicillin-clavulanic acid 95.0%, tetracycline 92.5%, and nitrofurantoin 80.0%. Otherwise, all isolates were susceptible to cefotaxime and ceftriaxone. All isolates were MDR, with 15 different profiles observed. Among 38 amoxicillin/clavulanic acid-resistant Salmonella isolates, 20 (52.6%) had the blaTEM gene, while blaSHV, blaCTX-M, and blaOXA genes were not detected. Only 5 (12.8%) out of 39 LEV-resistant isolates were positive for qnrB, three of which had blaTEM. No qnrC or qnrD, qnrS, aac(6`)-Ib-cr, qunA, and oqxAB genes were found in any of the tested isolates. Conclusion: This study demonstrates that broiler chickens may be considered a potential source for spreading MDR non-typhoidal Salmonella and ESBL traits in poultry production. Therefore, it is important to continuously monitor ESBL and PMQR genes to avoid the spread of resistant strains in the food chain and impact public health.

Polyaniline-Pectin nanoparticles immobilized paper based colorimetric sensor for detection of Escherichia coli in milk and milk products.

In the food quality and safety arena, there is a need to develop novel and sustainable methodologies that can help in the prevention of foodborne diseases. Herein, we report the development of a rapid conducting polymer strip-based sensor using Polyaniline-pectin (PANI-PEC) for the detection of Escherichia coli in milk and milk products. Polyaniline-pectin nanoparticles stabilized with biopolymer pectin were synthesized and its characterization studies such as FTIR, UV-Vis spectroscopy, electrical conductivity and particle size analysis were done. The assay parameters were optimized for the selective detection of E. coli in milk and milk products. The concentration of PANI-PEC solution immobilized/strip was optimized to be 3 mg/mL as it exhibited good sensitivity and colour intensity. Based on acid production and selectivity for E.coli, concentrations of media components like lactose, tryptophan, yeast extract, chondroitin sulphate, sodium lauryl sulphate, potassium chloride, tergitol-7, gentamycin sulphate and ampicillin trihydrate were optimized as 0.9, 0.1, 0.45, 0.015, 0.1, 2, 0.0125, 0.00016 and 0.015 respectively and sample volume was optimized to 500 µL. The developed PANI-PEC colorimetric strip-based sensor detects 0.52 ± 0.17 log CFU/mL E. coli within 10: 21 h (h). Further shelf-life study revealed that the developed PANI-PEC colorimetric sensor strips are stable at room temperature up to six months exhibiting the same sensitivity. The results obtained here indicate that this novel and simple paper based colorimetric sensor holds potential for application in food industries as a reliable and rapid method for detection of E. coli in milk and milk products at various stages of production and processing.

Development of an organic acid compound disinfectant to control food-borne pathogens and its application in chicken slaughterhouses.

During poultry slaughter, cross-contamination of chicken carcasses with microorganisms (including drug-resistant bacteria) can occur because of incomplete disinfection during the pre-cooling process, and surface contact with contaminated tools and equipment. The use of disinfectants is the most common way to reduce the risk of cross-contamination and bacterial spread, as they can effectively reduce the number of bacteria. We developed a disinfectant consisting of organic acids and sodium dodecyl sulfate (SDS) and tested its bactericidal effects at different concentrations against Salmonella and Campylobacter. The main effective components in the disinfectant were citric acid, lactic acid, and SDS, and together they exerted a synergistic bactericidal effect. The bactericidal efficacy of the disinfectant increased with increasing concentrations of the 3 active ingredients. To reach a 100% reduction rate during a 15-s treatment in vitro, for Salmonella, the lowest concentrations of citric acid, lactic acid, and SDS were 0.06, 0.08, and 0.02%, respectively; and for Campylobacter, the lowest concentrations were 0.02, 0.025, and 0.0125%, respectively. The disinfectant remained effective in presence of interfering substances (e.g., 15% fetal bovine serum). Further experiments showed that the disinfectant inactivated sensitive bacteria as well as 23 drug-resistant strains of Salmonella and Campylobacter. Treatment with the disinfectant for 15 s decreased the concentrations of all tested strains by more than 4.7 log colony forming units per mL, and the reduction rate was as high as 100%. In on-site disinfection tests in chicken slaughterhouses, the disinfectant significantly reduced the number of pathogenic bacteria on carcasses during the pre-cooling process, and on tools (such as knives and gloves) during the segmentation process. Thus, this disinfectant has potential uses in preventing cross-contamination of food-borne pathogens (including resistant bacteria) in slaughterhouses.

Establishment and application of a rapid visual detection method for Listeria monocytogenes based on polymerase spiral reaction (PSR).

Listeria monocytogenes is a common foodborne pathogen that presents in various food products, posing important threat to public health. The aim of this study was to establish a rapid and sensitive method with visualization to detect L. monocytogenes based on polymerase spiral reaction (PSR). Primers targeting conserved hlyA gene sequence of L. monocytogenes were designed based on bioinformatics analyses on the current available L. monocytogenes genomes. The isothermal amplification PSR can be completed under constant temperature (65ᵒC) within 60 min with high specificity and sensitivity. Twenty-five reference strains were used to evaluate the specificity of the developed reaction. The results showed that the sensitive of the reaction for L. monocytogenes in purified genomic DNA and artificially contaminated food samples were 41 pg/µL and 103 CFU/mL, respectively. It was 100-fold more sensitive than conventional PCR. In conclusion, this novel PSR method is rapid, cost-efficient, timesaving, and applicable on artificially contaminated food samples, providing broad prospects into the detection of foodborne microbes with the promising on-site inspection.

Isolation and characterization of E. coli O157: H7 novel bacteriophage for controlling this food-borne pathogen.

E. coli O157: H7 is known as a high-risk food-born pathogen, and its removal is vital for maintaining food safety. The increasing trend of food-borne diseases caused by this bacterium and other pathogens indicates the low efficiency of the methods to remove pathogens from foodstuffs. One of the new and effective methods is to use of a bio-control agent called bacteriophage, which has shown good function in eliminating and reducing pathogens. In this study, a novel bacteriophage was isolated and identified from the slaughterhouse wastewater to control E. coli O157: H7. This bacteriophage belonged to the Myoviridae family. Two bacterial genera including E. coli and Salmonella, were allocated to determine the bacteriophage host range; the result showed that the anti- Salmonella effect of phage was low. The phage was stable at high temperature (80 °C) and caused an acceptable reduction in the E. coli O157: H7 (4.18 log CFU / mL for 10 h). The isolated bacteriophage was corroborated to be completely safe based on the whole genome sequencing and lack of any virulence factor from the host bacteria. Considering the characteristics of this phage and its function in vitro, this bacteriophage may be used as an effective bio-control agent in foods with the possible E. coli O157: H7 -induced contamination.

'Three-To-One' multi-functional nanocomposite-based lateral flow immunoassay for label-free and dual-readout detection of pathogenic bacteria.

Sandwich lateral flow immunoassays (LFIAs) based on paired antibodies are the most frequently used platform for food-borne pathogen detection. Although label-free strategies are used in LFIAs to avoid the utilization of paired antibodies, challenges of probe design and detection reliability still remain. Here, we report a new label-free and dual-readout LFIA (LD-LFIA) mediated by a 'Three-To-One' multi-functional nanocomposite with a unique combination of magnetic-adhesion-color-nanozyme properties. The strengths of the new designed nanocomposite are: (i) the Fe3O4 magnetic core simplifies the separation processes; (ii) surface adherent polydopamine (PDA) films exhibit a strong adhesion to pathogenic bacteria and provide colorimetric detection signal; and (iii) the deposited platinum nanoparticles (Pt NPs) can function as nanozymes to generate an extra catalytic signal for constructing a dual-readout mode to improve the detection accuracy. The resulting Fe3O4@PDA@Pt nanocomposite-based LD-LFIA can detect highly pathogenic Escherichia coli O157:H7 with limits of detection of 102 and 10 CFU mL-1 for colorimetric and catalytic quantitative analyses, respectively. Systematic results also reveal that the proposed method exhibited high specificity and applicability for drinking water and chicken samples, serving as a promising tool for real bacterial sample testing. The multi-functional Fe3O4@PDA@Pt nanocomposite-based LD-LFIA can provide new ideas for designing new multi-functional probes for improving detection performance of conventional label-free LFIA and constructing more accurate and sensitive detection systems.

Bacillus cytotoxicus-A potentially virulent food-associated microbe.

Bacillus cytotoxicus is a member of the Bacillus cereus group with the ability to grow at high temperatures (up to 52℃) and to synthesize cytotoxin K-1, a diarrhoeagenic cytotoxin, which appears to be unique to this species and more cytotoxic than the cytotoxin K-2 produced by other members of this group. Only a few isolates of this species have been characterized with regard to their cytotoxic effects, and the role of cytotoxin K-1 as a causative agent of food poisoning remains largely unclear. Bacillus cytotoxicus was initially isolated from a food-borne outbreak, which led to three deaths, and the organism has since been linked to other outbreaks all involving plant-based food matrices. Other studies, as well as food-borne incidents reported to the UK Food Standards Agency, detected B. cytotoxicus in insect-related products and in dried food products. With insect-related food becoming increasingly popular, the association with this pathogen is concerning, requiring further investigation and evidence to protect public health. This review summarizes the current knowledge around B. cytotoxicus and highlights gaps in the literature from a food safety perspective.