Search Results (27)

Wild boars, widely distributed across natural, agricultural, and urban landscapes, represent an ideal sentinel species for monitoring the emergence and spread of antimicrobial resistance (AMR) at the human–wildlife–livestock interface within the One Health framework. This review summarizes current knowledge on the prevalence, diversity, AMR, and epidemiological significance of major enteric pathogens isolated from wild boars in the European Union, with particular attention to their potential role in AMR dissemination. Numerous studies have reported variable prevalence rates for Salmonella spp., Yersinia enterocolitica, Shiga toxin-producing Escherichia coli (STEC), and Campylobacter spp. High prevalence rates has been observed in fecal samples—35% for Salmonella, 27% for Y. enterocolitica and STEC, and 66% for Campylobacter—highlighting the role of wild boars as carriers and the associated risk of carcass contamination during slaughter. Tonsils represent a key niche for Y. enterocolitica, with prevalence reaching 35%. Several studies have identified resistance to antimicrobials classified by the World Health Organization as critically important or high priority for human medicine, including fluoroquinolone-resistant non-typhoidal Salmonella spp. and third-generation cephalosporin-resistant Y. enterocolitica, raising notable public health concerns. Despite increasing interest, most available studies remain descriptive and geographically limited, providing limited insight into AMR acquisition and transmission pathways in wild boars. New approaches—such as resistome analyses and epidemiological cut-off values—offer added value to distinguish wild-type from acquired-resistant strains and to better understand AMR dissemination dynamics. Integrating wildlife into One Health surveillance systems is essential to capture the full complexity of AMR spread. Full article

Shiga-toxin-producing E. coli (STEC) is a leading causing of bacterial foodborne and zoonotic illnesses in the USA. Whole-genome sequencing (WGS) is a powerful tool used in public health and microbiology for the detection, surveillance, and outbreak investigation of STEC. In this study, we applied three WGS-based subtyping methods, high quality single-nucleotide polymorphism (hqSNP) analysis, whole genome multi-locus sequence typing using chromosome-associated loci [wgMLST (chrom)], and core genome multi-locus sequence typing (cgMLST), to isolate sequences from 11 STEC outbreaks. For each outbreak, we evaluated the concordance between subtyping methods using pairwise genomic differences (number of SNPs or alleles), linear regression models, and tanglegrams. Pairwise genomic differences were highly concordant between methods for all but one outbreak, which was associated with international travel. The slopes of the regressions for hqSNP vs. allele differences were 0.432 (cgMLST) and 0.966 wgMLST (chrom); the slope was 1.914 for cgMLST vs. wgMLST (chrom) differences. Tanglegrams comprised of outbreak and sporadic sequences showed moderate clustering concordance between methods, where Baker’s Gamma Indices (BGIs) ranged between 0.35 and 0.99 and Cophenetic Correlation Coefficients (CCCs) were ≥0.88 across all outbreaks. The K-means analysis using the Silhouette method showed the clear separation of outbreak groups with average silhouette widths ≥0.87 across all methods. This study validates the use of cgMLST for the national surveillance of STEC illness clusters using the PulseNet 2.0 system and demonstrates that hqSNP or wgMLST can be used for further resolution. Full article

Biosensors are innovative and cost-effective analytical devices that integrate biological recognition elements (bioreceptors) with transducers to detect specific substances (biomolecules), providing a high sensitivity and specificity for the rapid and accurate point-of-care (POC) quantitative detection of selected biomolecules. In the meat production chain, their application has gained attention due to the increasing demand for enhanced food safety, quality assurance, food fraud detection, and regulatory compliance. Biosensors can detect foodborne pathogens (Salmonella, Campylobacter, Shiga-toxin-producing E. coli/STEC, L. monocytogenes, etc.), spoilage bacteria and indicators, contaminants (pesticides, dioxins, and mycotoxins), antibiotics, antimicrobial resistance genes, hormones (growth promoters and stress hormones), and metabolites (acute-phase proteins as inflammation markers) at different modules along the meat chain, from livestock farming to packaging in the farm-to-fork (F2F) continuum. By providing real-time data from the meat chain, biosensors enable early interventions, reducing the health risks (foodborne outbreaks) associated with contaminated meat/meat products or sub-standard meat products. Recent advancements in micro- and nanotechnology, microfluidics, and wireless communication have further enhanced the sensitivity, specificity, portability, and automation of biosensors, making them suitable for on-site field applications. The integration of biosensors with blockchain and Internet of Things (IoT) systems allows for acquired data integration and management, while their integration with artificial intelligence (AI) and machine learning (ML) enables rapid data processing, analytics, and input for risk assessment by competent authorities. This promotes transparency and traceability within the meat chain, fostering consumer trust and industry accountability. Despite biosensors’ promising potential, challenges such as scalability, reliability associated with the complexity of meat matrices, and regulatory approval are still the main challenges. This review provides a broad overview of the most relevant aspects of current state-of-the-art biosensors’ development, challenges, and opportunities for prospective applications and their regular use in meat safety and quality monitoring, clarifying further perspectives. Full article

Enteropathogenic (EPEC), necrotoxigenic (NTEC), and Shiga-toxin producing Escherichia coli (STEC) are pathotypes responsible for severe clinical forms in humans and animals. They can be shed in the feces of animals with consequent environmental contamination. This study evaluated the antibacterial activity of essential oils (EOs) from oregano (Origanum vulgare), savory (Satureja montana), thyme (Thymus vulgaris), and their blend against EPEC, NTEC, and STEC strains previously isolated from avian fecal samples. Minimum inhibitory concentration values between 0.039% and 0.156% were found with O. vulgare EO, between ≤0.0195% and 0.156% with both S. montana and T. vulgaris EOs, and between 0.039% and ≤0.0195% with the blend. The mixture with equal parts of EOs from oregano, savory and thyme seems a promising alternative product to combat pathogenic E. coli strains responsible for environmental contamination. Full article

Escherichia coli O157:H7, a Shiga-toxin-producing E. coli (STEC), is an important pathogen related to foodborne disease that is responsible for a growing number of outbreaks worldwide and has been detected in processed meats, dairy, and fresh vegetables. Although culturing is the gold standard method for detection of this bacterium, molecular methods based on nucleic acid amplification techniques such as PCR are becoming more common because of their rapidity, sensitivity, and specificity. However, to ensure reliable results among the several alternative PCR protocols (e.g., commercial kits and reference methods), different measurement assurance tools, including validated methods, reference materials, and proficiency tests, among others, are required. Herein, we present a digital PCR method validation for E. coli O157:H7 detection and quantification using seven specific gene sequences; this method quantified nucleic acids from different E. coli serotypes, with a detection range of 6.6 to 7900 copies/µL and a repeatability standard deviation over the concentration range of 1% to 13.6%. The relative standard uncertainty was 3.5–14.6%, and the detection limit was 0.27 copies/µL. Subsequently, two batches of a candidate reference material based on E. coli O157:H7 genomic DNA were then produced and characterized for evaluation of copy number concentration with the validated ddPCR method, with assigned values of 164,770 ± 9251 and 172 ± 9 copies/μL. Thus, this study demonstrated the development of a validated method and reference material for dPCR and qPCR detection of E. coli O157:H7, a key STEC responsible for food poisoning. Full article

The consumption of organic food has increased in recent years. In organic rearing animals are exposed to outdoor conditions, which may increase their risk of infection from various pathogens. In the present study the occurrence of the most significant foodborne pathogenic bacteria in organic meat and ready-to-eat organic meat products was assessed. Out of 100 raw organic meat samples tested, 72 were contaminated with bacterial pathogens. The highest percentage of contaminated samples was observed in poultry meat (92.5%) followed by pork meat (66.7%). Furthermore, 50.0% of beef origin samples were positive for the bacteria tested. L. monocytogenes was found in 39.0% of samples, S. aureus was identified in 37.0%, Campylobacter in 20.0%, Salmonella in 8.0% and Shigatoxin-producing E. coli in 4.0% of raw meat samples. In 31.0% of samples a co-occurrence of two (83.9%) or three (16.1%) pathogens was observed. Among 100 samples of organic meat products tested, only L. monocytogenes was found in 5.0% of samples. The result of the present study indicated that organic food may be a source of harmful microorganisms that may pose foodborne infections to consumers. Full article

Background: Foodborne diseases are a serious public health concern, and food-producing animals are a major source of contamination. Methods: The present study analysed Escherichia coli and Salmonella spp. isolated from faecal samples of 100 fattening pigs and from 52 samples of pork meat. Results: The results showed that the majority of the analysed meat samples were considered satisfactory in terms of microbiological quality (92.3% for E. coli and 94.2% for Salmonella spp.). Salmonella spp. was identified in 5.8% of the meat samples, whereas E. coli was detected in 89.5% of all samples (69.2% in meat and 100% in faecal samples). Furthermore, 1.9% of the faecal samples contained Shiga-toxin-producing E. coli and 3.9% contained enterotoxigenic E. coli. All sequenced isolates presented virulence genes for extraintestinal pathogenic E. coli. Moreover, 75.0% of E. coli isolates from meat and 71.8% from faeces samples showed antibiotic resistance, with 40.7% and 51.4%, respectively, being multidrug-resistant (MDR). The most prevalent resistances were to tetracycline, ampicillin, and sulfamethoxazole, and one E. coli isolate showed resistance to extended-spectrum β-lactamase. Conclusions: This study highlights the role of pigs as a potential source of human contamination and the importance of a One Health approach to ensure food safety and to promote public health. Full article

Shiga toxin (Stx), the main virulence factor of Shiga-toxin-producing E. coli (STEC), was first discovered in Shigella dysenteriae strains. While several other bacterial species have since been reported to produce Stx, STEC poses the most significant risk to human health due to its widespread prevalence across various animal hosts that have close contact with human populations. Based on its biochemical and molecular characteristics, Shiga toxin can be grouped into two types, Stx1 and Stx2, among which a variety of variants and subtypes have been identified in various bacteria and host species. Interestingly, the different Stx subtypes appear to vary in their host distribution characteristics and in the severity of diseases that they are associated with. As such, this review provides a comprehensive overview on the bacterial species that have been recorded to possess stx genes to date, with a specific focus on the various Stx subtype variants discovered in STEC, their prevalence in certain host species, and their disease-related characteristics. This review provides a better understanding of the Stx subtypes and highlights the need for rapid and accurate approaches to toxin subtyping for the proper evaluation of the health risks associated with Shiga-toxin-related bacterial food contamination and human infections. Full article

Wildlife can represent a reservoir of zoonotic pathogens and a public health problem. In the present study, we investigated the spread of zoonotic pathogens (Salmonella spp., Yersinia enterocolitica, Listeria monocytogenes, Shiga-toxin-producing Escherichia coli (STEC), and hepatitis E virus (HEV)) considering the presence of virulence and antibiotic resistance genes in game meat from animals hunted in northwest Italy. During two hunting seasons (2020 to 2022), samples of liver and/or muscle tissue were collected from chamois (n = 48), roe deer (n = 26), deer (n = 39), and wild boar (n = 35). Conventional microbiology and biomolecular methods were used for the detection, isolation, and characterization of the investigated pathogens. Two L. monocytogenes serotype IIa strains were isolated from wild boar liver; both presented fosfomycin resistance gene and a total of 22 virulence genes were detected and specified in the text. Eight Y. enterocolitica biotype 1A strains were isolated from chamois (2), wild boar (5), and deer (1) liver samples; all showed streptogramin and beta-lactam resistance genes; the virulence genes found were myfA (8/8 strains), ymoA (8/8), invA (8/8), ystB (8/8), and ail (4/8). Our data underscore the potential role of wildlife as a carrier of zoonotic and antibiotic-resistant pathogens in northwest Italy and a food safety risk for game meat consumers. Full article

Edema disease (ED) is a severe and lethal infectious ailment in swine, stemming from Shiga-toxin-producing Escherichia coli (STEC). An efficient, user-friendly, and safe vaccine against ED is urgently required to improve animal welfare and decrease antibiotic consumption. Recombinant attenuated Salmonella vaccines (RASV) administered orally induce both humoral and mucosal immune responses to the immunizing antigen. Their potential for inducing protective immunity against ED is significant through the delivery of STEC antigens. rSC0016 represents an enhanced recombinant attenuated vaccine vector designed for Salmonella enterica serotype Choleraesuis. It combines sopB mutations with a regulated delay system to strike a well-balanced equilibrium between host safety and immunogenicity. We generated recombinant vaccine strains, namely rSC0016 (pS-FedF) and rSC0016 (pS-rStx2eA), and assessed their safety and immunogenicity in vivo. The findings demonstrated that the mouse models immunized with rSC0016 (pS-FedF) and rSC0016 (pS-rStx2eA) generated substantial IgG antibody responses to FedF and rStx2eA, while also provoking robust mucosal and cellular immune responses against both FedF and rStx2eA. The protective impact of rSC0016 (pS-FedF) against Shiga-toxin-producing Escherichia coli surpassed that of rSC0016 (pS-rStx2eA), with percentages of 83.3%. These findings underscore that FedF has greater suitability for vaccine delivery via recombinant attenuated Salmonella vaccines (RASVs). Overall, this study provides a promising candidate vaccine for infection with STEC. Full article

Shiga-toxin-producing Escherichia coli (STEC) is typically detected on food products mainly due to cross-contamination with faecal matter. The serotype O157:H7 has been of major public health concern due to the severity of illness caused, prevalence, and management. In the food chain, the main methods of controlling contamination by foodborne pathogens often involve the application of antimicrobial agents, which are now becoming less efficient. There is a growing need for the development of new approaches to combat these pathogens, especially those that harbour antimicrobial resistant and virulent determinants. Strategies to also limit their presence on food contact surfaces and food matrices are needed to prevent their transmission. Recent studies have revealed that bacteriophages are useful non-antibiotic options for biocontrol of E. coli O157:H7 in both animals and humans. Phage biocontrol can significantly reduce E. coli O157:H7, thereby improving food safety. However, before being certified as potential biocontrol agents, the safety of the phage candidates must be resolved to satisfy regulatory standards, particularly regarding phage resistance, antigenic properties, and toxigenic properties. In this review, we provide a general description of the main virulence elements of E. coli O157:H7 and present detailed reports that support the proposals that phages infecting E. coli O157:H7 are potential biocontrol agents. This paper also outlines the mechanism of E. coli O157:H7 resistance to phages and the safety concerns associated with the use of phages as a biocontrol. Full article

Background: In the last decade, the importance of hetero-pathogenic enteroaggregative Shiga-toxin-producing E. coli for public health has increased. Recently, we described the genetic background of the EAHEC O181:H4 strain of ST678 carrying the stx2 gene in prophage and five plasmids, including the plasmid-carrying aggR and aaiC genes. Here, we present the morphological and enzymatic characteristics of this strain, as well as susceptibility to antimicrobials, biofilm formation, etc. Methods: Bacterial morphology was studied using an electron microscope. Susceptibility to antimicrobials was determined using the microdilution method. Cytotoxicity was estimated in Vero cells. Virulence was studied on mice. Results: The morphological and enzymatic properties of the hetero-pathogenic EAHEC strain were typical for E. coli; electron microscopy revealed the specific flagella. The strain was susceptible to most antibiotics and disinfectants but resistant to ampicillin and ciprofloxacin and showed a high degree of biofilm formation. Cytotoxicity towards Vero cells was estimated as 80%. Conclusions: The emergence of a new O181:H4 EAHEC strain poses a potential threat to humans because of the virulence potential that must be taken into account in the epidemiological analysis of outbreaks and sporadic cases of foodborne infections associated with hemolytic–uremic syndrome. Full article

The objective of this study was to evaluate the food safety efficacy of common antimicrobial interventions at and above required uptake levels for processing aids on the reduction of Shiga-toxin producing E. coli (STEC) and Salmonella spp. through spray and dip applications. Beef trim was inoculated with specific isolates of STEC or Salmonella strains. Trim was intervened with peracetic or lactic acid through spray or dip application. Meat rinses were serially diluted and plated following the drop dilution method; an enumerable range of 2–30 colonies was used to report results before log transformation. The combination of all treatments exhibits an average reduction rate of 0.16 LogCFU/g for STEC and Salmonella spp., suggesting that for every 1% increase in uptake there is an increase of 0.16 LogCFU/g of reduction rate. There is a statistical significance in the reduction rate of Shiga-toxin producing Escherichia coli in relation to the uptake percentage (p < 0.01). The addition of explanatory variables increases the R² of the regression for STEC, where all the additional explanatory variables are statistically significant for reduction (p < 0.01). The addition of explanatory variables increases the R² of the regression for Salmonella spp., but only trim type is statistically significant for reduction rate (p < 0.01). An increase in uptake percentages showed a significant increase in reduction rate of pathogens on beef trimmings. Full article

This study characterized the effect of distance from beef cattle feedlots, environmental factors, and climate on the occurrence of airborne bacterial indicators and pathogens. Three hundred air samples were collected over 6 months from five feedlots, with each air sample comprising 6000 L of air. Air samples were processed onto TSB-enriched air filters, qPCR-screened, and then qPCR-confirmed for suspect positive colonies of E. coli O157, non-O157-Shiga-toxin-producing E. coli (STEC), Salmonella, and E. coli. Direct enumeration of E. coli was also collected. Although no bacterial pathogens were qPCR-confirmed for the 300 samples, E. coli was detected in 16.7% (50/300) of samples, with an overall mean concentration of 0.17 CFU/6000 L air. Logistic regression analyses revealed a higher odds of E. coli for samples in close proximity compared to >610 m (2000 ft) distance from feedlots, along with significant associations with meteorological factors, sampling hour of day, and the presence of a dust-generating activity such as plowing a field or nearby vehicular traffic. The lack of bacterial pathogen detection suggests airborne deposition from nearby feedlots may not be a significant mechanism of leafy green bacterial pathogen contamination; the result of our study provides data to inform future revisions of produce-safety guidance. Full article

Lytic bacteriophages are re-considered as a solution to resolve antibiotic-resistant rampage. Despite frequent foodborne outbreaks caused by the top six non-O157 Shiga-toxin-producing Escherichia coli (STEC), the current interventions are not sufficiently effective against each serogroup, particularly O45. Therefore, this study aimed to characterize a new short-tailed phage, vB_EcoP-Ro45lw (or Ro45lw), as an alternative antimicrobial agent for STEC O45 strains. Phage Ro45lw belongs to the Kayfunavirus genus within the Autographiviridae family and shares no close evolutionary relationship with any reference phages. Ro45lw contains a tail structure composed of a unique tail fiber and tail tubular proteins A and B, likely to produce enzymatic activity against the target bacterial cells besides structural function. Additionally, the phage genome does not contain virulent, antibiotic-resistant, or lysogenic genes. The phage has a latent period of 15 min with an estimated burst size of 55 PFU/CFU and is stable at a wide range of pH (pH4 to pH11) and temperatures (30 °C to 60 °C). Regardless of the MOIs (MOI = 0.1, 1, and 10) used, Ro45lw has a strong antimicrobial activity against both environmental (E. coli O45:H-) and clinical (E. coli O45:H2) strains at 25 °C. These findings indicate that phage Ro45lw has antimicrobial potential in mitigating pathogenic STEC O45 strains. Full article