Enterohemorrhagic Research Articles

A novel shiga toxigenic E. coli sequence type harbouring multidrug resistance isolated from raw milk

Abstract The objective was to examine the prevalence of extended-spectrum β-lactam (ESBL) resistance among Shiga toxigenic Escherichia coli (STEC) isolated from raw milk. The findings from disc diffusion analysis and polymerase chain reaction revealed a high occurrence of ESBL resistance, specifically to penicillins, cephalosporins, and carbapenems, in isolated STEC strains. Additionally, a distinct sequence type of STEC was also identified in milk through comparative analysis.

Key roles of two-component systems in intestinal signal sensing and virulence regulation in enterohemorrhagic Escherichia coli.

Enterohemorrhagic Escherichia coli (EHEC) is a foodborne pathogen that infects humans by colonizing the large intestine. Upon reaching the large intestine, EHEC mediates local signal recognition and the transcriptional regulation of virulence genes to promote adherence and colonization in a highly site-specific manner. Two-component systems (TCSs) represent an important strategy used by EHEC to couple external stimuli with the regulation of gene expression, thereby allowing EHEC to rapidly adapt to changing environmental conditions. An increasing number of studies published in recent years have shown that EHEC senses a variety of host- and microbiota-derived signals present in the human intestinal tract and coordinates the expression of virulence genes via multiple TCS-mediated signal transduction pathways to initiate the disease-causing process. Here, we summarize how EHEC detects a wide range of intestinal signals and precisely regulates virulence gene expression through multiple signal transduction pathways during the initial stages of infection, with a particular emphasis on the key roles of TCSs. This review provides valuable insights into the importance of TCSs in EHEC pathogenesis, which have relevant implications for the development of antibacterial therapies against EHEC infection.

What, how, and why? - anti-EHEC phages and their application potential in medicine and food industry.

Enterohemorrhagic Escherichia coli (EHEC) are pathogens that, only in the United States, cause more than 250,000 foodborne infections a year. Since antibiotics or other antidiarrheal agents may increase the hemolytic-uremic syndrome (HUS) development risk, currently only supportive therapy, including hydration, is used. Therefore, many methods to fight EHEC bacteria focus on their use in food processing to prevent human infection. One of the proposed anti-EHEC agents is bacteriophages, known for their bactericidal effect, host specificity, and lack of cross-resistance with antibiotics. In this review article, we provide an overview of the characteristics like source of isolation, morphology, kinetics of life cycle, and treatment potential of over 130 bacteriophages able to infect EHEC strains. Based on the reviewed literature, we conclude that bacteriophages may play a highly significant role in regulating EHEC propagation. In addition, we also point out the phage features that should be taken into account not only when using bacteriophages but also when examining their properties. This may contribute to accelerating the pace of work on the preventive use of bacteriophages, which is extremely needed in the modern world of the food industry, but also stimulate interest in phages and accelerate regulatory work that would enable the use of bacteriophages also in medicine, to fight the drug-resistant strains.

Single Laboratory Evaluation of the Q20+ Nanopore Sequencing Kit for Bacterial Outbreak Investigations

Leafy greens are a significant source of produce-related Shiga toxin-producing Escherichia coli (STEC) outbreaks in the United States, with agricultural water often implicated as a potential source. Current FDA outbreak detection protocols are time-consuming and rely on sequencing methods performed in costly equipment. This study evaluated the potential of Oxford Nanopore Technologies (ONT) with Q20+ chemistry as a cost-effective, rapid, and accurate method for identifying and clustering foodborne pathogens. The study focuses on assessing whether ONT Q20+ technology could facilitate near real-time pathogen identification, including SNP differences, serotypes, and antimicrobial resistance genes. This pilot study evaluated different combinations of two DNA extraction methods (Maxwell RSC Cultured Cell DNA kit and Monarch high molecular weight extraction kits) and two ONT library preparation protocols (ligation and the rapid barcoding sequencing kit) using five well-characterized strains representing diverse foodborne pathogens. High-quality, closed bacterial genomes were obtained from all combinations of extraction and sequencing kits. However, variations in assembly length and genome completeness were observed, indicating the need for further optimization. In silico analyses demonstrated that Q20+ nanopore sequencing chemistry accurately identified species, genotype, and virulence factors, with comparable results to Illumina sequencing. Phylogenomic clustering showed that ONT assemblies clustered with reference genomes, though some indels and SNP differences were observed, likely due to sequencing and analysis methodologies rather than inherent genetic variation. Additionally, the study evaluated the impact of a change in the sampling rates from 4 kHz (260 bases pair second) to 5 kHz (400 bases pair second), finding no significant difference in sequencing accuracy. This evaluation workflow offers a framework for evaluating novel technologies for use in surveillance and foodborne outbreak investigations. Overall, the evaluation demonstrated the potential of ONT Q20+ nanopore sequencing chemistry to assist in identifying the correct strain during outbreak investigations. However, further research, validation studies, and optimization efforts are needed to address the observed limitations and fully realize the technology’s potential for improving public health outcomes and enabling more efficient responses to foodborne disease threats.

Genotypic analysis of Shiga toxin-producing Escherichia coli clonal complex 17 in England and Wales, 2014-2022.

Introduction. Shiga toxin-producing Escherichia coli (STEC) are zoonotic, gastrointestinal pathogens characterized by the presence of the Shiga toxin (stx) gene. Historically, STEC O157:H7 clonal complex (CC) 11 has been the most clinically significant serotype; however, recently there has been an increase in non-O157 STEC serotypes, including STEC O103:H2 belonging to CC17.Gap statement. STEC O103:H2 is an STEC serotype frequently isolated in England, although little is known about the epidemiology, clinical significance, associated public health burden or evolutionary context of this strain.Aim. Surveillance data and whole-genome sequencing data were analysed to determine the microbiological characteristics and public health burden of CC17, including the clinically significant serotype O103:H2, in England and Wales.Methodology. Isolates of E. coli belonging to CC17 (n=425) submitted to the Gastrointestinal Bacteria Reference Unit from 2014 to 2022 were whole genome sequenced, integrated with enhanced surveillance questionnaire data and analysed retrospectively.Results. Overall, diagnoses of CC17 infection increased every year since 2014. Most cases were female (58.5%), with the highest proportion of cases belonging to the 0-4 age group (n=83/424, 19.6%). Clinical presentation data identified diarrhoea (92.1%), abdominal pain (72.4%) and blood in stool (55.3%) as the most frequent symptoms, while 20.4% cases were admitted to hospital and 1.3% developed haemolytic uraemic syndrome. The five most common established serotypes were O103:H2 (64.5%), O123:H2 (11.1%), O151:H2 (6.6%), O71:H2 (3.3%) and O4:H2 (2.6%). The majority of CC17 isolates (78.6%) had the stx1a/eae virulence gene combination. Nine outbreak clusters of STEC infections that were mainly geographically dispersed and temporally related were identified and associated with foodborne transmission.Conclusions. Nationwide implementation of PCR to detect non-O157 STEC and improvements to algorithms for the follow-up of PCR-positive faecal specimens is recommended. Enhanced surveillance is necessary to assess the incidence of CC17 infection and overall burden of this CC within the UK population.

Cytoskeletal mechanisms regulating attaching/effacing bacteria interactions with host cells: It takes a village to build the pedestal.

The actin cytoskeleton is a key cellular structure subverted by pathogens to infect and survive in or on host cells. Several pathogenic strains of Escherichia coli, such as enteropathogenic E. coli (EPEC) and enterohemorrhagic E. coli (EHEC), developed a unique mechanism to remodel the actin cytoskeleton that involves the assembly of actin filament-rich pedestals beneath the bacterial attachment sites. Actin pedestal assembly is driven by bacterial effectors injected into the host cells, and this structure is important for EPEC and EHEC colonization. While the interplay between bacterial effectors and the actin polymerization machinery of host cells is well-understood, how other mechanisms of actin filament remodelling regulate pedestal assembly and bacterial attachment are poorly investigated. This review discusses the gaps in our understanding of the complexity of the actin cytoskeletal remodelling during EPEC and EHEC infection. We describe possible roles of actin depolymerizing, crosslinking and motor proteins in pedestal dynamics, and bacterial interactions with the host cells. We also discuss the biological significance of pedestal assembly for bacterial infection.

Gb3 trisaccharide-bearing exosomes as a novel neutralizer for Shiga toxin type 1

Shiga toxin types 1 (Stx1) and 2 (Stx2), produced by Shiga toxin-producing Escherichia coli (STEC) and Shigella dysenteriae, are key virulence factors responsible for severe foodborne diseases, such as hemorrhagic colitis and hemolytic uremic syndrome (HUS). The receptors for Stxs are Gb3 and P1 glycotope, which contain the Galα1→4Galβ epitope and are synthesized by human α1,4-galactosyltransferase (A4galt). Stx-related infections pose a global public health challenge, owing to the limited therapeutic options due to the restricted use of antibiotics. Therefore, there is an urgent need to develop novel therapeutic strategies. This study proposes an innovative strategy utilizing exosomes derived from CHO-Lec2 cells, which were modified with Functional-Spacer-Lipid (FSL) conjugates bearing the Gb3 carbohydrate epitope (exo-Gb3-FSL). Flow cytometry analysis confirmed the presence of Galα1→4Galβ disaccharides on exo-Gb3-FSL constructs, enabling them to bind Stx1. Moreover, using CHO-Lec2 cells verified the ability of exo-Gb3-FSL agents to bind Stx1 and protect these cells from Stx1-mediated cytotoxicity. For Stx1-treated CHO-Lec2 cells, increased cell survival was observed when using 25 μM exo-Gb3-FSL constructs, compared to control cells. These findings highlight the potential of exosome-based anti-Stx1 agents as promising alternatives to conventional therapies. This innovative strategy may provide novel directions for studies on Stx1 neutralization, offering a valuable strategy for the treatment of Stx-related diseases.

Isolation and molecular characterization of Shiga toxin-producing Escherichia coli (STEC) from bovine and porcine carcasses in Poland during 2019–2023 and comparison with strains from years 2014–2018

Isolation and molecular characterization of Shiga toxin-producing Escherichia coli (STEC) from bovine and porcine carcasses in Poland during 2019–2023 and comparison with strains from years 2014–2018

A novel multiplex and glycoprotein-based immunochromatographic serologic IgM test for the rapid diagnosis of Escherichia coli O157 and O145 causing bloody diarrhea and hemolytic uremic syndrome.

Shiga toxin-producing Escherichia coli (STEC) are the main etiological agents of hemolytic uremic syndrome (HUS). Good clinical management of STEC infections and HUS depends on early, rapid, and accurate diagnosis. Here, we have developed and evaluated the first multiplex and glycoprotein-based immunochromatographic test for the detection of IgM antibodies against the O-polysaccharide of the lipopolysaccharide of E. coli O157 and O145 in human serum samples. A retrospective study was carried out resulting in a diagnostic sensitivity of the E. coli O157/O145 LFIA (lateral flow immunoassay) of 97.1% and 98.9% for O157 and O145, respectively, and 97.9% for both serogroups. The diagnostic specificity was 98.7% for O157 and O145, and the overall specificity 97.4%. In samples obtained before 3 days after the onset of diarrhea, the detection percentage was 83%, increasing to 100% from 3 days onward. Finally, the association of bloody diarrhea (BD) or HUS cases to an STEC infection increased from 22.8% to 77.2% when stool culture and stx/Stx detection were combined with serology by LFIA. Our results demonstrate that the E. coli O157/O145 LFIA is a highly accurate and serospecific test for the early and rapid diagnosis of E. coli O157 and O145 infections in BD or HUS cases. This test allows the detection of specific IgM antibodies very early in the course of the infection, making it an ideal diagnostic tool to be implemented in pediatric emergencies and, thus, avoid delays in the application of the correct supportive or specific treatment and prevent complications associated with HUS.

Isolation, characterization, and receptor-binding protein specificity of phages PAS7, PAS59 and PAS61 infecting Shiga toxin-producing Escherichia coli O103 and O146

Shiga toxin-producing Escherichia coli (STEC) is a foodborne pathogen with 6,534 annual reported cases in the EU in 2021. This pathotype generally contains strains with smooth LPS with O-antigen serogroup O157 being the predominant serogroup in the US. However, non-O157 STEC serogroups are becoming increasingly prevalent. Here we announce the complete genomes of three newly isolated phages that infect STEC serogroups O103 and O146, namely Escherichia phages vB_EcoP_PAS7, vB_EcoP_PAS59 and vB_EcoP_PAS61. The genome sequences revealed that they belong to three distinct genera, namely the newly proposed genus Cepavirus within the Slopekvirinae subfamily, the genus Suseptimavirus and the genus Uetakevirus, respectively. We identified the tailspikes of phages PAS7 and PAS61 as a primary specificity determinant for the O-antigens O103 and O146, respectively, and predicted their active site in silico.

Chlorine dioxide is a broad-spectrum disinfectant against Shiga toxin-producing Escherichia coli and Listeria monocytogenes in agricultural water.

Agricultural water is commonly treated with chlorine-based disinfectants, which are impacted by water quality. Understanding how water quality influences disinfectants such as chlorine dioxide (ClO2) against pathogenic bacteria is important for creating efficacious sanitation regimens. In this study, the minimum inhibitory concentration (MIC) of ClO2 needed to achieve a 3-Log reduction against Shiga toxin-producing Escherichia coli (STEC) and Listeria monocytogenes was compared across agricultural water samples. Sterile ddH2O served as a control to compare with environmental samples from Salinas Valley, CA, and laboratory standards. To test different dosages and water qualities, stock ClO2 was diluted in 24-well plates with target concentrations of 10, 5, 2.5, and 1.25 mg/L. Well plates were inoculated with pathogens and treated with sanitizer for 5 min. Following treatment, surviving pathogens were enumerated using viable cell counts. The results demonstrate that groundwater samples had the highest water quality of the environmental samples and required the lowest concentration of disinfectant to achieve 3-Log reduction against both bacteria, with MIC between 1.4 and 2.0 mg/L. Open-source samples had lower water quality and required a higher concentration of ClO2 for 3-Log reduction, with MIC between 2.8 and 5.8 mg/L for both pathogens. There was no correlation between pH, turbidity, or conductivity/TDS and reduction for either STEC or L. monocytogenes, suggesting no individual water metric was driving reduction. A lower dosage was required to achieve 3-Log reduction against STEC, while L. monocytogenes required greater concentrations to achieve the same level of reduction. Overall, these results help guide growers in using ClO2 as a broad-spectrum disinfectant and demonstrate its efficacy in reaching 3-Log reduction across agricultural water samples.

Shiga toxin-producing Escherichia coli in the milk production chain: Evaluation of virulence genes and clonal diversity of O157:H7 strains

Microbial contamination of milk and dairy products is a significant issue due to sensory changes and the risk of foodborne diseases. Shiga toxin-producing Escherichia coli (STEC), particularly the O157:H7, are major pathogens transmitted by dairy products. This study aimed to evaluate the microbiological quality of bovine milk at various stages of the production chain in a dairy cooperative and its suppliers in Rio de Janeiro, Brazil. Additionally, the presence of STEC in milk and faeces from dairy cattle was investigated, along with the isolation of STEC O157:H7 strains and the evaluation of virulence genes and clonal diversity. These findings were compared with previously isolated STEC O157:H7 strains from the same region. A total of 100 raw milk samples (54 farm gallon (FG) samples, 25 community bulk tank milk (CBTM) samples, 14 isothermal car-tanker milk (ICTM) samples, and 7 industry tank milk (ITM) samples) and 10 pasteurized milk samples, along with 63 bovine faeces samples, were analysed. The stx gene, a marker for STEC, was detected in 65% of raw milk samples and 88.9% of faecal samples, indicating a high risk of contamination throughout the production chain. None of the pasteurized milk samples tested positive for the stx gene. The prevalence of stx in faeces and raw milk underscores the need for rigorous controls to prevent contamination. While pasteurization was effective, the microbiological quality of raw milk remained unsatisfactory, highlighting the importance of good hygiene practices and continuous monitoring to ensure dairy safety.

Isolation and evaluation of the pathogenicity of a hybrid shiga toxin-producing and Enterotoxigenic Escherichia coli in pigs.

Porcine pathogenic Escherichia coli (E. coli), the globally recognized important pathogen, causes significant economic loss in the field. Enterotoxigenic E. coli (ETEC) causes porcine neonatal and post-weaning diarrhea (PWD), frequently carrying F4 adhesin, F18 adhesin, Heat-Stable toxin (ST), and Heat-Labile toxin (LT). Shiga Toxin-Producing E. coli (STEC) produces F18 adhesin and Shiga toxin type 2e (stx2e), majorly leading to systemic endothelial cell damage and edema disease. In this study, hemolytic pathogenic hybrid STEC/ETEC strains carrying ST and LT genes of ETEC and the Stx2e gene of STEC isolated from pigs with PWD in Taiwan were identified. The pathogenicity of a Taiwan hybrid STEC/ETEC strain was evaluated by oral inoculation in post-weaning pigs. Next generation sequencing and multilocus sequence typing of two hybrid Taiwan porcine STEC/ETEC isolates indicated that these two isolates were closely related to the ST88 porcine hybrid STEC/ETEC isolated from pigs with watery diarrhea. Furthermore, the two hybrid Taiwan porcine STEC/ETEC isolates also displayed combinations of multiple resistance genes encoding mechanisms for target modification and antibiotic inactivation. Animal experiments confirmed that the Taiwan hybrid STEC/ETEC could cause watery diarrhea in post-weaning pigs with no signs of edema disease and minimal histopathological lesions. To the best of the authors' knowledge, the present study is the first study demonstrating intestinal pathogenicity of the hybrid STEC/ETEC in pigs. The result suggests that the hybrid STEC/ETEC should be considered as a new emerging pathogen and a new target for vaccine development.

The inactivation of Shiga toxin-producing Escherichia coli (STEC) and Listeria monocytogenes using isochoric freezing in raw milk and carrot juice

Isochoric freezing is a novel food preservation method that may maintain the quality and safety of products by simultaneously applying low temperatures and high pressures. This work aims to determine whether isochoric freezing can improve food safety by inactivating Shiga toxin-producing Escherichia coli (STEC) and Listeria monocytogenes in raw milk and carrot juices. Raw milk and carrot juice samples were inoculated with cocktails of either STEC or L. monocytogenes and then subjected to isochoric freezing conditions in pressurized chambers set at −5, −10, and −15 °C and treated for 1, 5, 7, or 10 days. Following treatment, the product was sampled for surviving pathogens. The results demonstrated that a 5-log reduction of both STEC and L. monocytogenes can be achieved for raw milk and carrot juice without adversely affecting quality. For STEC inoculated in raw milk, both the linear and Weibull models suggest that 10 days is required to achieve 5-log reduction at −10 °C. Decreasing the temperature to −15 °C led to accelerated log reduction but induced phase separation in the milk. Listeria in raw milk demonstrated a biphasic reduction, indicating 1.3 days is required for a 5-log reduction at −10 °C. In comparison, pathogens demonstrated faster log reduction in carrot juice due to its intrinsic properties. The survival curves for STEC-inoculated carrot juice demonstrated that the 5-log reduction times were 6.9 days at −10 °C and 3.8 days at −15 °C. L. monocytogenes in carrot juice required 1.5 days at −10 °C for a 5-log reduction and was eliminated (7 log cfu/ml) within 24 h at −15 °C. Isochoric freezing could be an option for milk and carrot juice processors wishing to improve food safety without applying heat. This method is not a one-size-fits-all solution, and the time and temperature will need further optimization depending on the target pathogen and intended commodity.

A master regulator of central carbon metabolism directly activates virulence gene expression in attaching and effacing pathogens.

The ability of the attaching and effacing pathogens enterohaemorrhagic Escherichia coli (EHEC) and Citrobacter rodentium to overcome colonisation resistance is reliant on a type 3 secretion system used to intimately attach to the colonic epithelium. This crucial virulence factor is encoded on a pathogenicity island known as the Locus of Enterocyte Effacement (LEE) but its expression is regulated by several core-genome encoded transcription factors. Here, we unveil that the core transcription factor PdhR, traditionally known as a regulator of central metabolism in response to cellular pyruvate levels, is a key activator of the LEE. Through genetic and molecular analyses, we demonstrate that PdhR directly binds to a specific motif within the LEE master regulatory region, thus activating type 3 secretion directly and enhancing host cell adhesion. Deletion of pdhR in EHEC significantly impacted the transcription of hundreds of genes, with pathogenesis and protein secretion emerging as the most affected functional categories. Furthermore, in vivo studies using C. rodentium, a murine model for EHEC infection, revealed that PdhR is essential for effective host colonization and maximal LEE expression within the host. Our findings provide new insights into the complex regulatory networks governing bacterial pathogenesis. This research highlights the intricate relationship between virulence and metabolic processes in attaching and effacing pathogens, demonstrating how core transcriptional regulators can be co-opted to control virulence factor expression in tandem with the cell's essential metabolic circuitry.

Investigation of Shiga toxin-producing Escherichia coli and evaluation of the microbiological quality of raw vegetable sal-ads

Foodborne diseases (FBD) are a significant public health concern worldwide, affecting millions of people annually. Symptoms of FBD range from mild gastrointestinal discomfort to more severe conditions, which in some cases can lead to death. Raw vegetables, especially those consumed in ready-to-eat (RTE) salads, are particularly susceptible to microbial contamination. This study aimed to investigate the presence of Shiga toxin-producing Escherichia coli (STEC) and to evaluate the microbiological quality of 77 RTE raw vegetable salads from 20 restaurants in Niterói, RJ, Brazil. None samples presented STEC, nor the safety indicators (SI) Salmonella spp. and Listeria monocytogenes. However, 89.6% of the samples were considered unfit for human consumption, since they presented at least one hygiene indicator (HI) outside the established standards. The majority of samples (95.5%) were unsatisfactory for Enterobacteriaceae (EB), followed by total aerobic bacteria (TAB) (76.6%), thermotolerant coliforms (C45) (32.5%) and Escherichia coli (EC) (1.3%). The difference between the occurrences of hygiene indicators was not significant. Given the high percentage of inadequate samples found in the study, it is clear that there is a need for stricter surveillance and adequate hygiene practices in restaurants. To reduce the risks of microbial contamination in RTE raw vegetable salads, several approaches can be implemented. These include continuous training of food handlers, implementation of food safety management systems such as Hazard Analysis and Critical Control Points (HACCP), and regular inspections. In addition, consumers also play an important role by demanding safe food and following safe food handling and consumption practices at home.

Exploring the Intersection of Atypical Hemolytic Uremic Syndrome and Substance Use: A Comprehensive Narrative Review.

Hemolytic uremic syndrome (HUS) is a thrombotic microangiopathy characterized by hemolytic anemia, renal failure, and thrombocytopenia. While the typical form of HUS is often associated with Shiga toxin-producing Escherichia coli (STEC) infections, atypical hemolytic uremic syndrome (aHUS) is caused by uncontrolled complement system activation, leading to endothelial damage, microthrombi formation, and other complications. Although aHUS is commonly linked to genetic mutations and infections, emerging evidence suggests that certain substances, particularly illicit drugs like heroin, cocaine, and ecstasy, can also trigger this condition, adding complexity to its diagnosis and management. This narrative review examines the mechanisms by which substance use can lead to aHUS, discusses its clinical presentation, and highlights the diagnostic challenges in distinguishing it from other thrombotic microangiopathies, such as thrombotic thrombocytopenic purpura (TTP) and STEC-HUS. A thorough literature search identified relevant case reports, case series, and observational studies, underscoring the need for genetic testing and complement assays to confirm aHUS in substance users. The review also explores the role of complement inhibitors, such as eculizumab and ravulizumab, which target the underlying pathophysiology and have shown promise in improving patient outcomes. However, the management of substance-induced aHUS remains challenging due to limited data, varying clinical presentations, and the need to optimize treatment protocols. Early recognition and tailored therapy are crucial for effective management. Further research is needed to refine diagnostic criteria, develop new therapeutic approaches, and improve care for patients with this under-recognized condition.

Prevalence and characteristics of 11 potentially diarrhoeagenic microbes in asymptomatic individuals in Norway, 2015-2020.

We aimed to estimate the prevalence of potentially diarrhoeagenic microbes (PDMs) in faecal samples from asymptomatic individuals in a high-income country, identify risk factors for carriage and to identify microbial factors that differ between PDMs in asymptomatic versus symptomatic individuals. Samples from 1000 asymptomatic participants were collected, together with a questionnaire, between 2015 and 2020 and examined by PCR for 11 PDMs. Isolates were characterised and potential risk factors were registered. Atypical enteropathogenic Escherichia coli (aEPEC), Yersinia enterocolitica, Shiga toxin-producing E. coli (STEC), enterotoxigenic E. coli (ETEC) and Campylobacter spp. were found in 163 (16%), 20 (2.0%), 17 (1.7%), 12 (1.2%) and 11 (1.1%) asymptomatic individuals, respectively. Other PDMs were rare. Only low virulent STEC, with stx1c, stx2b or stx2f, was detected. Travels outside Europe was a significant risk factor for detecting Campylobacter spp. (odds ratio (OR) 6.99; 95% CI 1.12-43.6) and ETEC (OR 11.4; 95% CI 1.26-102). Individuals ≥65 years of age had lower odds of carrying STEC (OR 0.11; 95% CI 0.02-0.57) or EPEC (OR 0.09; 95% CI 0.05-0.16) than individuals ≤5 years of age. The common finding of PDMs in asymptomatic individuals could have implications for the interpretation of positive findings in clinical samples and infection control measures.

Commercial bacteriophage preparations for the control of Listeria monocytogenes and Shiga toxin-producing Escherichia coli in raw and pasteurized milk

Listeria monocytogenes was the etiologic agent in nearly all recent outbreaks in North America attributed to pasteurized dairy products, whereas Escherichia coli O157 infections were responsible for most of the rare, yet serious complications from outbreaks involving unpasteurized dairy. This study determined the susceptibility of selected strains of L. monocytogenes and Shiga toxin-producing E. coli (STEC) to commercial phage preparations and their ability to control these pathogens in pasteurized and raw milk during 7-day storage at 7 °C. Both phage products demonstrated high lytic efficiency against 17 strains of L. monocytogenes whereas the efficiency of E. coli phages was more variable against 11 strains of O157 and non-O157 STEC. Broth microdilution assays identified effective endpoint multiplicities of infection (MOI) ranging from log 2.53 to 5.13, which differed between strains of L. monocytogenes and phage products. Mean log MOIs of E. coli phages against STEC also varied within and between products from 0.43 to 7.05. Despite these observations, the change in counts over time of three L. monocytogenes strains exposed to phage in pasteurized milk (log MOI 6) was similar with counts ∼ 4 log CFU/mL lower than control at day 7. Results for STEC O157 varied by strain but counts were lower than control in all cases by 72 h thorough day 7. Titers on isolates of both pathogens isolated from pasteurized milk indicated that the surviving populations were less susceptible to phage. The addition of a phage preparation to raw milk did not reduce populations of either pathogen or affect the change in counts of any strain over time when compared to control. Reduced efficacy in raw milk may be attributed to reduced phage binding as titers in raw milk decreased steadily (∼2 log PFU/mL) during storage. Commercial phage products may be a promising pathogen control intervention for pasteurized dairy products, warranting further investigation.

Host-Pathogen Interactions during Shiga Toxin-Producing Escherichia coli Adherence and Colonization in the Bovine Gut: A Comprehensive Review.

Shiga toxin-producing Escherichia coli (STEC) is a significant public health threat due to its ability to cause severe gastrointestinal diseases in humans, ranging from diarrhea to life-threatening conditions such as hemorrhagic colitis and hemolytic uremic syndrome (HUS). As the primary reservoir of STEC, cattle play a crucial role in its transmission through contaminated food and water, posing a considerable risk to human health. This comprehensive review explores host-pathogen interactions during STEC colonization of the bovine gut, focusing on the role of gut microbiota in modulating these interactions and influencing disease outcomes. We integrated findings from published transcriptomics, proteomics, and genomics studies to provide a thorough understanding of how STEC adheres to and colonizes the bovine gastrointestinal tract. The insights from this review offer potential avenues for the development of novel preventative and therapeutic strategies aimed at controlling STEC colonization in cattle, thereby reducing the risk of zoonotic transmission.