Shiga Toxin-producing Escherichia Coli Infections Research Articles

Growth behavior of Shiga toxin-producing Escherichia coli, Salmonella, and generic E. coli in raw pork considering background microbiota at 10, 25, and 40 °C

Recent epidemiological evidence suggests that pork products may be vehicles for the transmission of Shiga toxin-producing Escherichia coli (STEC) to humans. The severe morbidity associated with STEC infections highlights the need for research to understand the growth behavior of these bacteria in pork products. Classical predictive models can estimate pathogen growth in sterile meat. However, competition models considering background microbiota reflect a more realistic scenario for raw meat products. The objective of this study was to estimate the growth kinetics of clinically significant STEC (O157, non-O157, and O91), Salmonella, and generic E. coli in raw ground pork using competition primary growth models at temperature abuse (10 and 25 °C) and sublethal temperature (40 °C). A competition model incorporating the No lag Buchanan model was validated using the acceptable prediction zone (APZ) method where >92 % (1498/1620) of the residual errors fell within the APZ (pAPZ > 0.70). The background microbiota (mesophilic aerobic plate counts, APC) inhibited the growth of STEC and Salmonella indicating a simple one-directional competitive interaction between pathogens and the mesophilic microbiota of ground pork. The maximum specific growth rate (μmax) of all the bacterial groups was not significantly different (p > 0.05) based on fat content (5 vs 25 %) except for generic E. coli at 10 °C. E. coli O157 and non-O157 behaved similarly in terms of μmax and maximum population density (MPD). Salmonella showed a similar (p > 0.05) μmax to E. coli O157 and non-O157 at 10 and 40 °C but a significantly higher rate (p < 0.05) at 25 °C. STEC were more prone to be inhibited by APC than Salmonella at 10 and 25 °C. The μmax of O91 was lower (p < 0.05) than other STEC and Salmonella at 10 and 25 °C but similar (p > 0.05) at 40 °C. Generic E. coli showed a two- to five-times higher (p < 0.05) μmax (0.028 ± 0.011 log10 CFU/h) than other bacterial groups (0.006 ± 0.004 to 0.012 ± 0.003 log10 CFU/h) at 10 °C making it a potential indicator bacteria for process control. Industry and regulators can use competitive models to develop appropriate risk assessment and mitigation strategies to improve the microbiological safety of raw pork products.

Core Genome Sequencing Analysis of E. coli O157:H7 Unravelling Genetic Relatedness among Strains from Cattle, Beef, and Humans in Bishoftu, Ethiopia

E. coli O157:H7 is a known Shiga toxin-producing Escherichia coli (STEC), causing foodborne disease globally. Cattle are the main reservoir and consumption of beef and beef products contaminated with E. coli O157:H7 is an important source of STEC infections in humans. To emphasize the cattle-to-human transmission through the consumption of contaminated beef in Bishoftu, Ethiopia, whole-genome sequencing (WGS) was performed on E. coli O157 strains isolated from three sources (cattle, beef, and humans). Forty-four E. coli O157:H7 isolates originating from 23 cattle rectal contents, three cattle hides, five beef carcasses, seven beef cuts at retail shops, and six human stools in Bishoftu between June 2017 and May 2019 were included. This study identified six clusters of closely related E. coli O157:H7 isolates based on core genome multilocus sequence typing (cgMLST) by targeting 2513 loci. A genetic linkage was observed among the isolate genomes from the cattle rectal contents, cattle hides, beef carcasses at slaughterhouses, beef at retail shops, and human stool within a time frame of 20 months. All the strains carried practically the same repertoire of virulence genes except for the stx2 gene, which was present in all but eight of the closely related isolates. All the strains carried the mdfA gene, encoding for the MdfA multi-drug efflux pump. CgMLST analysis revealed genetically linked E. coli O157:H7 isolates circulating in the area, with a potential transmission from cattle to humans through the consumption of contaminated beef and beef products.

Diarrhoea-associated haemolytic uraemic syndrome and Shiga toxin-producing Escherichia coli infections in New Zealand children: Clinical features and short-term complications from a 23-year cohort study.

Diarrhoea-associated haemolytic uraemic syndrome (D+HUS) is an important cause of acute kidney injury (AKI) in young children and it is most commonly associated with Shiga toxin-producing Escherichia coli (STEC). Gastrointestinal infections caused by STEC have been increasing in New Zealand over the past two decades, but little is known regarding the acute and short-term outcomes of New Zealand children who develop D+HUS. To describe the clinical characteristics, complications and short-term outcomes of New Zealand children with D+HUS identified between 1 January 1998 and 31 December 2020. The New Zealand Paediatric Surveillance Unit sends out a monthly survey to all practising paediatricians regarding conditions under active surveillance. Paediatricians caring for a child aged 0-15 years of age with D+HUS over the prior month were requested to report their patient. Reporting clinicians were then contacted by the principal investigator and sent a questionnaire requesting patient clinical and laboratory information. Two hundred and twenty-six children had D+HUS; median age 2.8 years (interquartile range 1.7-4.9). Acute dialysis was required in 128/226 (56.2%) of children for a median of 9 days (range 1-38). Children with shorter diarrhoeal prodrome, higher neutrophil count and haemoglobin had a longer duration of dialysis. Seizures occurred in 31/226 (13.7%) and were not associated with a greater HUS severity score. Acute mortality was 1.3%, all resulting from thrombotic microangiopathic cerebral injury. D+HUS is a major cause of AKI in previously healthy young children. Earlier recognition of STEC infections in young children may reduce the need for dialysis and other extra-renal complications. The New Zealand incidence of acute dialysis, other major complications and mortality are consistent with other reported studies.

Burden of disease estimation based on Escherichia coli quantification in ready-to-eat meals served in Portuguese institutional canteens

Shiga toxin-producing E. coli (STEC), enterotoxigenic E. coli (ETEC) and enteropathogenic E. coli (EPEC) are associated with the onset of gastroenteritis with different severities, and STEC has been associated with other sequelae, such as hemolytic uremic syndrome and end stage renal disease. The main goal of this study was to estimate the annual foodborne burden of disease associated with STEC, ETEC and EPEC infection based on E. coli quantification results obtained in the routine analysis of ready-to-eat meals served in institutional canteens from 2018 to 2019. A stochastic Quantitative Microbial Risk Assessment (QMRA) model was used to estimate the expected number of cases per health outcome and Disability Adjusted Life Years (DALYs). Assuming a daily consumption of a whole meal portion (450 g), the estimated burden was of 4.2 x 10−3 DALYs/person/year for STEC infection, 2.82 x 10−4 DALYs/person/year for ETEC infection and 7.91 x 10−6 DALYs/person/year for EPEC infection. Additionally, using the Sobol method, the sensitivity analysis revealed that the factors with higher influence on the final output (DALYs) were the pathotype's prevalence for the STEC model, the number of people exposed to the hazard for the ETEC model and E. coli concentrations for the EPEC model.

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.

Shiga toxin (stx) encoding genes in sheep and goats reared in Trinidad and Tobago.

Shiga toxin-producing Escherichia coli (STEC) is estimated to cause over two million cases of human disease annually. Trinidad and Tobago is one of the largest livestock producer and consumer of sheep and goat meat in the Caribbean, however, the potential role of these animals in the epidemiology of STEC infections has not been previously described. To fill this critical gap in knowledge, the prevalence of Shiga toxin genes (stx1 and stx2) shed in the faeces of healthy sheep (n = 204) and goats (n = 105) in Trinidad was investigated. Based on PCR screening, goats had a higher stx prevalence than sheep (46% vs 35%, P = 0.06). Most of the recovered STEC isolates were positive for stx1 only; and only three isolates were positive for the eae gene. None of the recovered isolates belonged to the O157 serogroup. In both species, the prevalence of stx was higher in young animals versus older animals. Sheep reared on deep litter flooring (43%) had a higher prevalence than sheep reared other flooring types, however this was not the same for goats. The presence of cows on the same premise was not an associated predictor for STEC carriage in sheep or goats. This study demonstrates that although sheep and goats in Trinidad are reservoirs for stx-positive E. coli isolates, no fecal samples tested positive for O157 STEC, harbored. Furthermore, it appears that non-O157 stx-positive isolates harbored by these animals do not pose a significant threat to human health.

A Multi-Specific DARPin Potently Neutralizes Shiga Toxin 2 via Simultaneous Modulation of Both Toxin Subunits.

Shiga toxin-producing E. coli (STEC) is a common cause of bloody diarrhea. The pathology of STEC infection derives from two exotoxins—Shiga toxin 1 (Stx1) and Shiga toxin 2 (Stx2)—that are secreted by STEC in the gut, from where they are systemically absorbed, causing severe kidney damage leading to hemolytic uremic syndrome (HUS). Currently, there is no effective treatment for HUS, and only supportive care is recommended. We report the engineering of a panel of designed ankyrin repeat proteins (DARPin) with potent neutralization activity against Stx2a, the major subtype associated with HUS. The best dimeric DARPin, SD5, created via a combination of directed evolution and rational design, neutralizes Stx2a with a half maximal effective concentration (EC50) of 0.61 nM in vitro. The two monomeric DARPin constituents of SD5 exhibit complementary functions—SHT targets the enzymatic A subunit of Stx2a and inhibits the toxin’s catalytic activity, while DARPin #3 binds the B subunit, based on the cryo-EM study, and induces a novel conformational change in the B subunit that distorts its five-fold symmetry and presumably interferes with toxin attachment to target cells. SD5 was fused to an albumin-binding DARPin, and the resulting trimeric DARPin DA1-SD5 efficiently protects mice in a toxin challenge model, pointing to a high potential of this DARPin as a therapeutic for STEC infection. Finally, the unprecedented toxin conformational change induced by DARPin #3 represents a novel mode of action for neutralizing Stx2 toxicity and reveals new targets for future drug development.

Plasmon-Enhanced Bimodal Nanosensors: An Enzyme-Free Signal Amplification Strategy for Ultrasensitive Detection of Pathogens.

Increasing foodborne illnesses have led to global health and economic burdens. E. coli O157:H7 is one of the most common disease-provoking pathogens and known to be lethal Shiga toxin-producing E. coli (STEC) strains. With a low infection dose in addition to person-to-person transmission, STEC infections are easily spread. As a result, specific and rapid testing methods to identify foodborne pathogens are urgently needed. Nanozymes have emerged as enzyme-mimetic nanoparticles, demonstrating intrinsic catalytic activity that could allow for rapid, specific, and accurate pathogen identification in the agrifood industry. In this study, we developed a sensitive nanoplatform based on the traditional ELISA assay with the synergistic properties of gold and iron oxide nanozymes, replacing the conventional enzyme horseradish peroxidase (HRP). We designed an easily interchangeable sandwich ELISA composed of a novel, multifunctional magneto-plasmonic nanosensor (MPnS) with target antibodies (MPnS-Ab). Our experiments demonstrate a 100-fold increase in catalytic activity in comparison to HRP with observable color changes within 15 min. Results further indicate that the MPnS-Ab is highly specific for E. coli O157:H7. Additionally, effective translatability of catalytic activity of the MPnS technology in the lateral flow assay (LFA) platform is also demonstrated for E. coli O157:H7 detection. As nanozymes display more stability, tunable activity, and multi-functionality than natural enzymes, our platform could provide customizable, low-cost assay that combines high specificity with rapid detection for a variety of pathogens in a point-of-care setup.

Atypical Presentation of Shiga Toxin Haemolytic Uremic Syndrome (STEC-HUS)

Shiga-toxin producing Escherichia coli (STEC) infection is the most common cause of haemolytic uremic syndrome (HUS) in children. It should be noted that 5% of STEC-HUS patients have no prodromal diarrhoea, while it may be present in up to 30% of atypical HUS (aHUS) cases. In patients at risk for aHUS, STEC infection may act as a trigger rather than playing a causative role. Presently, eculizumab is the first-line therapy for children presenting with aHUS, whereas treatment of children with STEC-HUS is mainly supportive. We report the case of a child with STEC-HUS who had a severe presentation, requiring renal replacement therapy in the acute phase and surgical treatment for a colonic stricture that arose as a late extra-renal complication of the disease. We aim to review the diagnostic workup of children presenting with HUS, stressing the resources available in our setting. In Portugal, O157 E. coli isolation medium is widely available in clinical laboratories. Non-O157 STEC identification requires specific testing, which is increasingly relevant as non-O157 STEC is becoming a more common cause of STEC-HUS than serogroup O157. The National Health Institute Doutor Ricardo Jorge (INSA) has the capacity to identify verotoxin genes and to proceed to verotxin-producing E. coli (VTEC) pathogenicity assessment using multiplex polymerase chain reaction. Fresh stool samples must be obtained early in the disease course to be sent for culture in O157 E. coli isolation medium and for non-O157 E. coli identification assays.

Pre-treatment with phages achieved greater protection of mice against infection with Shiga toxin-producing Escherichia coli than post-treatment

Shiga toxin-producing Escherichia coli (STEC) is a group of pathogen that can cause various diseases in both humans and animals, such as watery diarrhea, hemorrhagic colitis, and uremia syndrome. Due to the serious situation of antibiotic resistance, phage therapy is considered to have a great potential in combating bacterial diseases. In this study, three phages (NJ-10, NJ-20, and NJ-38) with strong abilities to lyse virulent STEC strain CVCC193 cells in vitro were isolated. Subsequently, the therapeutic effects of the three phages were investigated in mice infected with CVCC193 cells. The results showed that the survival rates of mice injected with the phages at 3 h after challenge with CVCC193 cells were 40%–50%, while the survival rates of mice injected with the phages at 24 h before challenge were 80%–100%, indicating that pre-treatment with phages had better therapeutic effects than post-treatment. Pathological changes, bacterial loads in different organs, and serum levels of inflammatory factors of the infected mice were also detected. The results showed that the mice injected with the phages at 3 h after or 24 h before challenge with CVCC193 cells had significantly decreased organ lesions, bacterial loads, and serum levels of inflammatory factors as compared to infected mice without phage treatment. These results suggested that phages NJ-10, NJ-20, and NJ-38 can potentially protect against STEC infections.

Comparative Genomics of Shiga Toxin-Producing Escherichia coli Strains Isolated from Pediatric Patients with and without Hemolytic Uremic Syndrome from 2000 to 2016 in Finland.

ABSTRACTShiga toxin-producing Escherichia coli (STEC) infection can cause mild to severe illness, such as nonbloody or bloody diarrhea, and the fatal hemolytic uremic syndrome (HUS). The molecular mechanism underlying the variable pathogenicity of STEC infection is not fully defined so far. Here, we performed a comparative genomics study on a large collection of clinical STEC strains collected from STEC-infected pediatric patients with and without HUS in Finland over a 16-year period, aiming to identify the bacterial genetic factors that can predict the risk to cause HUS and poor renal outcome. Of 240 STEC strains included in this study, 52 (21.7%) were from pediatric patients with HUS. Serotype O157:H7 was the main cause of HUS, and Shiga toxin gene subtype stx2a was significantly associated with HUS. Comparative genomics and pangenome-wide association studies identified a number of virulence and accessory genes overrepresented in HUS-associated STEC compared to non-HUS STEC strains, including genes encoding cytolethal distending toxins, type III secretion system effectors, adherence factors, etc. No virulence or accessory gene was significantly associated with risk factors for poor renal outcome among HUS patients assessed in this study, including need for and duration of dialysis, presence and duration of anuria, and leukocyte counts. Whole-genome phylogeny and multiple-correspondence analysis of pangenomes could not separate HUS STEC from non-HUS STEC strains, suggesting that STEC strains with diverse genetic backgrounds may independently acquire genetic elements that determine their varied pathogenicity. Our findings indicate that nonbacterial factors, i.e., characteristics of the host immunity, might affect STEC virulence and clinical outcomes.IMPORTANCE Shiga toxin-producing Escherichia coli (STEC) is a serious public health burden worldwide which causes outbreaks of gastrointestinal diseases and the fatal hemolytic uremic syndrome (HUS) characterized by the triad of mechanical hemolytic anemia, thrombocytopenia, and acute renal failure. Understanding the mechanism underlying the disease severity and patient outcome is of high importance. Using comparative genomics on a large collection of clinical STEC strains from STEC-infected patients with and without HUS, our study provides a reference of STEC genetic factors/variants that can be used as predictors of the development of HUS, which will aid risk assessment at the early stage of STEC infection. Additionally, our findings suggest that nonbacterial factors may play a primary role in the renal outcome in STEC-infected patients with HUS; further studies are needed to validate this.

"FAMILY CLUSTER ASSOCIATED WITH SHIGA TOXIN-PRODUCING ESCHERICHIA COLI O26 SEROGROUP"

"Introduction: Shiga toxin-producing Escherichia coli (STEC) can cause severe human infections especially in the pediatric population who are at risk of progressing to life threatening systemic complications such as hemolytic uremic syndrome (HUS). In Romania, enhanced monitoring of STEC-associated HUS was implemented in year 2016 after a cluster of infections caused by STEC O26:H11 serotype heightened public awareness of their consequences. Objectives: This study describes the microbiological investigation initiated to identify a STEC-associated HUS pediatric case and determine additional infections among his household contacts. Methods: Fecal specimens collected from a small child hospitalized with HUS and his asymptomatic parents and brother were investigated using a culture-based approach which targeted STEC isolation, identification, and typing. PCR-based assays for key virulence genes (i.e., stx1 and stx2 types/subtypes, eae, and ehxA) were combined with traditional methods such as serogroup determination by slide agglutination. Also, multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE) were used for strain genotyping. Results: STEC was identified only in the stool samples of the siblings, one who developed HUS and the other who remained asymptomatic. Both strains belonged to O26 E. coli serogroup and displayed an identical virulence gene profile (presence of stx2a, eae, and ehxA genes). Molecular typing by PFGE showed the STEC strains as highly related genetically and MLST assigned them to sequence type ST21 lineage. Conclusion: A family cluster of O26 STEC infections was detected by using a culture- based molecular approach. Further analysis at whole genome sequencing resolution level is needed for a detailed characterization of such pathogens of significant public health importance. Keywords: HUS, Shiga toxin-producing Escherichia coli, virulence genes, PCR, molecular typing"

The Importance of Shiga Toxin-Producing Escherichia coli O145:NM[H28]/H28 Infections in Argentina, 1998-2020.

Shiga toxin-producing Escherichia coli (STEC) is known as a pathogen associated with food-borne diseases. The STEC O145 serogroup has been related with acute watery diarrhea, bloody diarrhea, hemorrhagic colitis, and hemolytic uremic syndrome (HUS). Argentina has the highest rate of HUS worldwide with 70% of the cases associated with STEC infections. We aimed to describe the epidemiology and genetic diversity of STEC O145 strains isolated across Argentina between 1998–2020. The strains isolated from 543 cases of human disease and four cattle, were pheno-genotipically characterized. Sequencing of five strains was performed. The strains were serotyped as O145:NM[H28]/H28, O145:H25, and O145:HNT, and mainly characterized as O145:NM[H28]/stx2a/eae/ehxA (98.1%). The results obtained by sequencing were consistent with those obtained by traditional methods and additional genes involved in different mechanisms of the pathogen were observed. In this study, we confirmed that STEC O145 strains are the second serogroup after O157 and represent 20.3% of HUS cases in Argentina. The frequency of STEC O145 and other significant serogroups is of utmost importance for public health in the country. This study encourages the improvement of the surveillance system to prevent severe cases of human disease.

Microbial risk assessment of Escherichia coli shiga-toxin producers (STEC) in raw sheep's milk cheeses in Italy

Shiga toxin-producing E. coli (STEC), widespread pathogens associated with severe foodborne disease, can contaminate milk during the milking process through faecal matter and survive or grow during cheese making if a pasteurization treatment has not been applied. Thus, a stochastic “farm-to-fork” model was developed to assess the risk of human infection by O157 STEC, one of the main pathogenic serotypes, associated with the consumption of a portion of raw sheep's milk cheese produced in a farmhouse dairy in Italy. The average risk of illness after the consumption of a portion of brief-, medium- and long-ripened cheese ranged between 1.64 × 10−4 and 4.03 × 10−4 for adults. Considering only a difference in serving size, the risk for children varied from 1.35 × 10−4 to 3.34 × 10−4. Among the several intervention strategies simulated to mitigate the risk, administration of bacteriophages was, by far, the most effective measure with an average risk reduction of 34 times followed by use of probiotics and antimicrobials, which lowered the risk about 12 times. The sensitivity analysis showed that the probability that a shedder is present in the herd, the occurrence of the milk contamination with faeces and the within-herd prevalence of the pathogen were the parameters that most affected the risk. While further data is necessary to confirm the conclusion of this study, the model results might be able to assist producers and policymakers to manage the risk of STEC infection linked to such products.

Shiga Toxin-Producing Escherichia coli and Milk Fat Globules.

Shiga toxin-producing Escherichia coli (STEC) are zoonotic Gram-negative bacteria. While raw milk cheese consumption is healthful, contamination with pathogens such as STEC can occur due to poor hygiene practices at the farm level. STEC infections cause mild to serious symptoms in humans. The raw milk cheese-making process concentrates certain milk macromolecules such as proteins and milk fat globules (MFGs), allowing the intrinsic beneficial and pathogenic microflora to continue to thrive. MFGs are surrounded by a biological membrane, the milk fat globule membrane (MFGM), which has a globally positive health effect, including inhibition of pathogen adhesion. In this review, we provide an update on the adhesion between STEC and raw MFGs and highlight the consequences of this interaction in terms of food safety, pathogen detection, and therapeutic development.

Interventions for Shiga toxin-producing Escherichia coli gastroenteritis and risk of hemolytic uremic syndrome: A population-based matched case control study.

The role of antibiotics in the treatment of Shiga toxin-producing Escherichia coli (STEC) infection is controversial. To evaluate the association between treatment (antibiotics, antidiarrheal agents, and probiotics) for STEC infection and hemolytic uremic syndrome (HUS) development. We performed a population-based matched case-control study using the data from the National Epidemiological Surveillance of Infectious Diseases (NESID) between January 1, 2017 and December 31, 2018. We identified all patients with STEC infection and HUS as cases and matched patients with STEC infection without HUS as controls, with a case-control a ratio of 1:5. Further medical information was obtained by a standardized questionnaire. Multivariable conditional logistic regression model was used. 7760 patients with STEC infection were registered in the NESID. 182 patients with HUS and 910 matched controls without HUS were selected. 90 patients with HUS (68 children and 22 adults) and 371 patients without HUS (266 children and 105 adults) were included in the main analysis. The matched ORs of any antibiotics and fosfomycin for HUS in children were 0.56 (95% CI 0.32-0.98), 0.58 (0.34-1.01). The matched ORs for HUS were 2.07 (1.07-4.03), 0.86 (0.46-1.61) in all ages treated with antidiarrheal agent and probiotics. Antibiotics, especially fosfomycin, may prevent the development of HUS in children, while use of antidiarrheal agents should be avoided.

AB5 Enterotoxin-Mediated Pathogenesis: Perspectives Gleaned from Shiga Toxins.

Foodborne diseases affect an estimated 600 million people worldwide annually, with the majority of these illnesses caused by Norovirus, Vibrio, Listeria, Campylobacter, Salmonella, and Escherichia coli. To elicit infections in humans, bacterial pathogens express a combination of virulence factors and toxins. AB5 toxins are an example of such toxins that can cause various clinical manifestations, including dehydration, diarrhea, kidney damage, hemorrhagic colitis, and hemolytic uremic syndrome (HUS). Treatment of most bacterial foodborne illnesses consists of fluid replacement and antibiotics. However, antibiotics are not recommended for infections caused by Shiga toxin-producing E. coli (STEC) because of the increased risk of HUS development, although there are conflicting views and results in this regard. Lack of effective treatment strategies for STEC infections pose a public health threat during outbreaks; therefore, the debate on antibiotic use for STEC infections could be further explored, along with investigations into antibiotic alternatives. The overall goal of this review is to provide a succinct summary on the mechanisms of action and the pathogenesis of AB5 and related toxins, as expressed by bacterial foodborne pathogens, with a primary focus on Shiga toxins (Stx). The role of Stx in human STEC disease, detection methodologies, and available treatment options are also briefly discussed.

Improved Genomic Identification, Clustering, and Serotyping of Shiga Toxin-Producing Escherichia coli Using Cluster/Serotype-Specific Gene Markers.

Shiga toxin-producing Escherichia coli (STEC) have more than 470 serotypes. The well-known STEC O157:H7 serotype is a leading cause of STEC infections in humans. However, the incidence of non-O157:H7 STEC serotypes associated with foodborne outbreaks and human infections has increased in recent years. Current detection and serotyping assays are focusing on O157 and top six (“Big six”) non-O157 STEC serogroups. In this study, we performed phylogenetic analysis of nearly 41,000 publicly available STEC genomes representing 460 different STEC serotypes and identified 19 major and 229 minor STEC clusters. STEC cluster-specific gene markers were then identified through comparative genomic analysis. We further identified serotype-specific gene markers for the top 10 most frequent non-O157:H7 STEC serotypes. The cluster or serotype specific gene markers had 99.54% accuracy and more than 97.25% specificity when tested using 38,534 STEC and 14,216 non-STEC E. coli genomes, respectively. In addition, we developed a freely available in silico serotyping pipeline named STECFinder that combined these robust gene markers with established E. coli serotype specific O and H antigen genes and stx genes for accurate identification, cluster determination and serotyping of STEC. STECFinder can assign 99.85% and 99.83% of 38,534 STEC isolates to STEC clusters using assembled genomes and Illumina reads respectively and can simultaneously predict stx subtypes and STEC serotypes. Using shotgun metagenomic sequencing reads of STEC spiked food samples from a published study, we demonstrated that STECFinder can detect the spiked STEC serotypes, accurately. The cluster/serotype-specific gene markers could also be adapted for culture independent typing, facilitating rapid STEC typing. STECFinder is available as an installable package (https://github.com/LanLab/STECFinder) and will be useful for in silico STEC cluster identification and serotyping using genome data.

The epidemiology of Shiga toxin-producing Escherichia coli serogroup O157 in England, 2009-2019.

Shiga toxin-producing Escherichia coli (STEC) serogroup O157 is a zoonotic, foodborne gastrointestinal pathogen of major public health concern. We describe the epidemiology of STEC O157 infection in England by exploring the microbiological and clinical characteristics, the demographic and geographical distribution of cases, and examining changes in environmental exposures over 11 years of enhanced surveillance. Enhanced surveillance data including microbiological subtyping, clinical presentations and exposures were extracted for all cases resident in England with evidence of STEC O157 infection, either due to faecal culture or serology detection. Incidence rates were calculated based on mid-year population estimates from the Office of National Statistics (ONS). Demographics, geography, severity and environmental exposures were compared across the time periods 2009-2014 and 2015-2019. The number of cases reported to national surveillance decreased, with the mean cases per year dropping from 887 for the period 2009-2014 to 595 for the period 2015-2019. The decline in STEC O157 infections appears to be mirrored by the decrease in cases infected with phage type 21/28. Although the percentage of cases that developed HUS decreased, the percentage of cases reporting bloody diarrhoea and hospitalisation remained stable. The number of outbreaks declined over time, although more refined typing methods linked more cases to each outbreak. Integration of epidemiological data with microbiological typing data is essential to understanding the changes in the burden of STEC infection, assessment of the risks to public health, and the prediction and mitigation of emerging threats.

High prevalence and pathogenic potential of Shiga toxin-producing Escherichia coli strains in raw mutton and beef in Shandong, China

Shiga toxin-producing Escherichia coli (STEC) is a foodborne pathogen that can cause severe human diseases such as hemolytic uremic syndrome (HUS). Human STEC infections are frequently caused through consumption of contaminated foods, especially raw meats. This study aimed to investigate the prevalence of STEC in raw meats and to characterize the meat-derived STEC strains using whole genome sequencing. Our study showed that 26.6% of raw mutton, and 7.5% of raw beef samples were culture-positive for STEC. Thirteen serotypes were identified in 22 meat-derived isolates in this study, including the virulent serotypes O157:H7 and O26:H11. Seven Shiga toxin (Stx) subtypes were found in 22 isolates, of these, stx1c and stx1c + stx2b were predominant. The recently-reported stx2k subtype was found in three mutton-sourced isolates. A number of other virulence genes such as genes encoding intimin (eae), enterohemorrhagic E. coli (EHEC) hemolysin (ehxA), EHEC factor for adherence (efa1), heat-stable enterotoxin 1 (astA), type III secretion system effectors, were detected in meat-derived STEC strains. One mutton-sourced isolate was resistant to three antibiotics, i.e., tetracycline, chloramphenicol, and trimethoprim-sulfamethoxazole. Whole-genome phylogeny indicated the genomic diversity of meat-derived strains in this study. O157:H7 and O26:H11 isolates in this study were phylogenetically grouped together with strains from HUS patients, suggesting their pathogenic potential. To conclude, our study reported high STEC contaminations in retail raw meats, particularly raw mutton, genomic characterization indicated pathogenic potential of meat-derived STEC strains. These findings highlight the critical need for increased monitoring of STEC in retail raw meats in China.