Shiga Toxin-producing Escherichia Coli Infections Research Articles

Shiga toxin-producing Escherichia coli infection-related acute encephalopathy.

Shiga toxin-producing Escherichia coli (STEC) poses a significant global health threat, contributing to gastrointestinal involvement and, rarely, systemic illnesses. In particular, the toxin's pathogenic mechanisms primarily target the kidneys and the central nervous system, leading to severe consequences. We present a rare case of STEC-related acute encephalopathy in a 34-year-old female without concurrent kidney involvement. The patient presented with hemorrhagic diarrhea and subsequent neurological symptoms, including seizures, cortical blindness, and left hemiparesis. Diagnostic evaluations, including blood tests, imaging studies, and cerebrospinal fluid analysis, suggested STEC infection-related encephalopathy. Central nervous system involvement in STEC infections, although more commonly observed in children, can manifest in various neurological symptoms, including altered mental status, seizures, and focal neurological deficits. Neuroimaging and electrophysiological findings often demonstrate non-specific abnormalities, reflecting diffuse brain dysfunction. The pathogenesis of STEC-related encephalopathy involves direct neuronal cytotoxicity mediated by Shiga toxin, leading to acute cerebrovascular events and neuroinflammation. Therapeutic strategies primarily focus on anti-inflammatory interventions, such as high-dose steroid therapy, with variable prognosis influenced by age, sex, disease severity, and response to treatment. This case underscores the importance of recognizing STEC infection-related encephalopathy as a rare but potentially severe complication, highlighting the need for prompt diagnosis and appropriate management to optimize patient outcomes.

P-130. A Case-Control Study of Post-Infectious Sequelae Following a Shiga Toxin-Producing Escherichia coli (STEC) Outbreak at a U.S. Marine Corps Recruit Depot

Abstract Background In November 2017, an outbreak of Shiga toxin-producing Escherichia coli (STEC) O157:H7 and O26 occurred among male recruits at a United States Marine Corps Recruit Depot, San Diego, CA. We sought to evaluate the 6-year risk of hypertension (HTN), chronic kidney disease (CKD), and irritable bowel syndrome (IBS) following STEC infection by conducting a retrospective, case-control study utilizing medical record data.Table 1.Proportion of STEC cases and matched asymptomatic controls with post-infectious sequelae following November 2017 outbreak Methods Data from the outbreak investigation were used to identify cases and asymptomatic controls. Asymptomatic controls were recruits who were in the exposure area during the outbreak but remained asymptomatic and were negative for STEC by stool PCR. Cases were matched 1:4 with asymptomatic controls based on the duration of follow-up available in the Military Health System Data Repository (MDR). Controls could be matched to > 1 case. Post-infectious outcomes were assessed using ICD-10 diagnostic codes in the MDR. Results 239 cases were identified of whom 192 (80%) developed bloody diarrhea, 30 (13%) were hospitalized, and 17 (7%) developed hemolytic uremic syndrome (HUS). The stx-2 gene was detected in 39% of PCR positive cases. The median age of recruits was 19 years, and the median duration of MDR follow-up for cases and controls was 3.7 years (IQR: 2.9-5.4). There was no significant difference in the proportion of cases and controls with post-infectious sequelae (Table 1). Resolution of renal dysfunction was noted within 3 months after discharge for all HUS cases, and only 1 case remained hypertensive following discharge. There was no significant difference in the proportion of bloody diarrhea and HUS cases that developed IBS compared to controls. A limitation of the analyses was the short duration of MDR follow-up available for 6/30 hospitalized cases due to military separation after hospital discharge. Conclusion STEC infection in this cohort of young military recruits was associated with a good prognosis, even among cases of STEC-associated HUS, with resolution of renal dysfunction and no increase in incidence of IBS or HTN compared with asymptomatic controls. The findings suggest that STEC infection in young adults who recover from acute illness is not associated with a higher likelihood of post-infectious sequelae. Disclosures Ryan C. Maves, MD, AiCuris: Grant/Research Support|Biotest: Grant/Research Support|GeoVax: Grant/Research Support|Shionogi: Advisor/Consultant|Shionogi: Honoraria|Sound Pharmaceuticals: Grant/Research Support

Virulent Bacteriophages for Controlling Shiga Toxin-Producing Escherichia coli (STEC) Without Inducing Toxin Production.

Shiga toxin-producing Escherichia coli (STEC) infections pose a significant public health challenge, characterized by severe complications including hemolytic uremic syndrome (HUS) due to Shiga toxin (Stx) production. Current therapeutic approaches encounter a critical limitation, as conventional antibiotic treatment is contraindicated due to its propensity to trigger bacterial SOS response and subsequently enhance Stx production, which increases the likelihood of developing HUS in antibiotic-treated patients. The lack of effective, safe therapeutic options has created an urgent need for alternative treatment strategies for STEC infections. This study investigates the therapeutic potential of virulent bacteriophages (phages) against STEC O157:H7. Our findings demonstrate that these phages effectively reduce STEC populations to levels comparable to ciprofloxacin treatment, while crucially avoiding the induction of SOS response and subsequent enhancement of Stx production. This is a significant advantage over conventional antibiotics which increase Stx levels. Furthermore, these phages exhibited broad-spectrum antimicrobial activity against multiple clinical STEC isolates without triggering toxin expression. The capability of virulent phages to effectively control STEC without enhancing toxin production represents a promising therapeutic strategy that combines antimicrobial efficacy with safety considerations. These characteristics indicate that virulent phages represent a potential solution to address the current therapeutic challenges in STEC infections, particularly in mitigating the risk of HUS development in infected patients.

Ruminant-dense environments increase risk of reported Shiga toxin-producing Escherichia coli infections independently of ruminant contact.

Cattle and other domestic ruminants are the primary reservoirs of O157 and non-O157 Shiga toxin-producing Escherichia coli (STEC). Living in areas with high ruminant density has been associated with excess risk of infection, which could be due to both direct ruminant contact and residual environmental risk, but the role of each is unclear. We investigated whether there is any meaningful risk to individuals living in ruminant-dense areas if they do not have direct contact with ruminants. Using a Bayesian spatial framework, we investigated the association between the density of ruminants on feedlots and STEC incidence in Minnesota from 2010 to 2019, stratified by serogroup and season, and adjusting for direct ruminant contact. For every additional head of cattle or sheep per 10 acres, the incidence of O157 STEC infection increased by 30% (incidence rate ratio [IRR] 1.30; 95% credible interval [CrI] 1.18, 1.42) or 135% (IRR 2.35; 95% CrI 1.14, 4.20), respectively, during the summer months. Sheep density was also associated with O157 STEC risk during winter (IRR 4.28; 95% CrI 1.40, 8.92). The risk of non-O157 STEC infection was only elevated in areas with goat operations during summer (IRR 19.6; 95% CrI 1.69, 78.8). STEC risk associated with ruminant density was independent of direct ruminant contact across serogroups and seasons. Our findings demonstrate that living in a ruminant-dense area increases an individual's risk of O157 and non-O157 STEC infection even without direct ruminant contact, indicating that prevention efforts need to extend to community strategies for averting indirect transmission from local ruminant populations.IMPORTANCEShiga toxin-producing Escherichia coli (STEC) are zoonotic enteric bacteria responsible for 2.5 million illnesses each year. Infections in young children can be especially devastating, causing hemolytic uremic syndrome (HUS), a debilitating and sometimes fatal form of acute kidney injury. STEC's primary reservoirs are cattle and other domestic ruminants, and transmission can occur through food, water, animal contact, and person-to-person. Living near ruminants poses a significant risk of STEC infection; however, the proportion of that risk due to direct ruminant contact or other routes of transmission is unknown. Our research demonstrates that direct ruminant contact is a substantial risk irrespective of location, and that individuals living in ruminant-rich regions are at high risk of STEC infection regardless of whether they come into contact with ruminants. These findings indicate a need for multi-pronged prevention efforts that emphasize control of contamination in the environments surrounding ruminant populations, in addition to biosafety precautions when contacting ruminants directly.

Characterization of Broad Spectrum Bacteriophage vB ESM-pEJ01 and Its Antimicrobial Efficacy Against Shiga Toxin-Producing Escherichia coli in Green Juice.

Shiga toxin-producing Escherichia coli (STEC) infections have increased in humans, animals, and the food industry, with ready-to-eat (RTE) food products being particularly susceptible to contamination. The prevalence of multidrug-resistant strains has rendered the current control strategies insufficient to effectively control STEC infections. Herein, we characterized the newly isolated STEC phage vB_ESM-pEJ01, a polyvalent phage capable of infecting Escherichia and Salmonella species, and assessed its efficacy in reducing STEC in vitro and food matrices. The phage, belonging to the Tevenvirinae, exhibits effective bacteriolytic activity, a short latent period, large burst size, and stability under a broad pH range and moderate temperatures. Moreover, the phage demonstrated strong anti-biofilm efficacy even at low concentrations. Genomic analysis revealed that the phage was similar to the well-characterized RB49 phage (T4-like phage) but possesses distinct host-specificity-related genes that potentially contribute to its extensive host range. The efficacy of phage vB_ESM-pEJ01 was evaluated in artificially STEC-inoculated green juice samples, where it significantly reduced STEC and the abundance of Shiga toxin-producing genes at 4 and 25 °C. Therefore, these results suggest that the polyvalent phage vB_ESM-pEJ01 is a promising biocontrol agent for foodborne pathogens in RTE foods such as fresh juices.

Epidemiology of Shiga toxin-producing Escherichia coli other than serotype O157:H7 in England, 2016-2023.

Introduction. Shiga toxin-producing Escherichia coli (STEC) infections are of public health concern as STEC can cause large national foodborne outbreaks of severe gastrointestinal disease, particularly in the young and elderly. In recent years, the implementation of PCR by diagnostic microbiology laboratories has improved the detection of STEC, and there has been an increase in notifications of cases of non-O157 STEC. However, the extent this increase in caseload can be attributed to the improved detection by PCR, or a true increase in non-O157 STEC infections, is unknown.Gap Statement. Epidemiological and microbiological data and analyses describing the trends in non-O157 STEC in England since the implementation of PCR are limited.Aim. Demographic, microbiological and clinical characteristics of non-O157 STEC from 8 years (2016-2023) of laboratory surveillance data were analysed to understand the recent trends in non-O157 serotypes and the incidence of disease in England.Methodology. All human isolates of STEC non-O157 detected between 2016 and 2023 were extracted from the laboratory surveillance system. Microbiological data were analysed and linked to clinical outcomes.Results. There was an almost 10-fold increase in diagnoses of non-O157 STEC from 2016 (n=297) to 2023 (n=2341). A total of 9378 isolates of non-O157 STEC were detected, comprising 338 different serotypes, and were linked to 9311 individuals. A higher proportion of non-O157 STEC cases were female (56%) and aged between 20 and 39 years (27%). The most common non-O157 serotypes were O26:H11 (16%), O146:H21 (12%), O91:H14 (11%), O128:H2 (6%), O145:H28 (5%) and O103:H2 (4%). STEC O26:H11 was more frequently reported in under 5s than any other age group (38%), whereas the other common serotypes were more frequently isolated from adults. Stx2a, which has been associated with greater disease severity, was detected in 18% of cases. Where clinical details were available, 27% of non-O157 cases were admitted to the hospital and 6% developed HUS. Cases of STEC O145:H28 reported a higher rate of hospitalisation than other non-O157 STEC cases. The serotypes most likely to be associated with progression to HUS were O26:H11 (9%) and O145:H28 (7%). STEC harbouring stx2f (19%), stx2a (11%) and stx2d (11%) were most frequently isolated from cases with HUS.Conclusion. The implementation of widespread PCR testing in England has facilitated better surveillance of STEC non-O157, with respect to establishing the true incidence and burden of disease of non-O157 STEC and monitoring the emergence of highly virulent strains.

Serogroups and Toxin Variants of Clinical Shiga Toxin-Producing Escherichia coli Strains Isolated from Patients with Hemolytic Uremic Syndrome in Turkey.

Shiga toxin-producing Escherichia coli (STEC) refers to a group of bacteria that can cause infections, which are common worldwide and pose a serious public health problem, as they can lead to conditions such as hemorrhagic colitis and hemolytic uremic syndrome (HUS). HUS is a disease characterized by microangiopathic hemolytic anemia, thrombocytopenia, and renal failure. Determination of serogroups and toxin profiles of STEC is important for estimating their disease-causing potential and predicting epidemiological changes. This study analyzed STEC isolates from 46 pediatric HUS patients across Turkey, using polymerase chain reaction to determine O serogroups and Shiga toxin (Stx) variants from stool samples collected between 2016 and 2019. Of the patients, 25 (54.3%) were in the 0-2 age group. Of the isolates, 82.6% were non-O157 serogroup. The most detected serogroup was O145 (32.6%), and 28.3% of the serogroups were not typed. Of the strains, 8 (17.4%) had Stx1 alone, 26 (56.5%) had Stx2 alone, and 12 (26.1%) had both Stx1 and Stx2. The Stx variants occurred in seven combinations, with the most common being Stx2a alone (56.5%). The duration of hospitalization for patients with Stx2a was found to be longer than that for patients with other variants (p = 0.01). This study highlights a concerning trend in Turkey, where non-O157 serogroups, particularly O145, emerged as prevalent causes of HUS. The predominance of Stx2a among our isolates and the longer hospitalization duration for patients with Stx2a support findings linking this variant to severe clinical outcomes, including HUS. Understanding the dynamics of these Stx variants will help better prepare for and mitigate the impacts of STEC infections in our population.

How colonizing alfalfa sprouts modulates the virulence of Shiga toxin-producing Escherichia coli

Shiga toxin-producing Escherichia coli (STEC), a significant cause of foodborne illnesses, is often associated with the consumption of fresh produce, including alfalfa sprouts. This study was executed to determine how quickly STEC grows, adapts, and colonizes alfalfa sprouts during production and storage, and whether the pathogen's virulence and infectious doses are affected by physiological adaptation to sprouts as an environment. A reporter STEC O157:H7 EDL933 strain was developed to track the transcription of eae, a virulence gene involved in colonizing human intestinal enterocytes. When the seeds were inoculated with 2.1 × 103 CFU/g of the reporter strain, the pathogen's population increased to 1.5 × 106 CFU/g sprouts within 1.38 days and then remained stable during the remainder of the 5-day sprouting, indicating physiological adaptation to this environment. Seeds were inoculated with ∼108 CFU/g and subsequently treated with 2000 ppm calcium hypochlorite solution, followed by a water-rinse (treated seeds), or just rinsed with water (untreated seeds). After 5 days of sprouting, the resulting fresh sprouts were refrigerated for three days at 4 °C. Sprout samples were collected and treated with 2000 ppm calcium hypochlorite solution and rinsed thoroughly with water before counting internalized STEC, or just water-washed before measuring total STEC. The transcription of eae (normalized to cell count) was the highest on the second day of sprouting, but the transcription of other virulence and stress-related genes varied, with sodA being upregulated in STEC cells. Lethal dose 50 (LD50) to Galleria mellonella, a STEC infection animal model, was lower (i.e., virulence was higher) in total STEC collected from fresh sprouts produced from treated seeds, compared to that from untreated seeds (1.9 × 100 and 6.0 × 101 CFU/larva, respectively). Compared to refrigerated sprouts, the LD50 of STEC from freshly produced sprouts was lower. Based on these findings, it can be concluded that (a) STEC quickly adapts physiologically to sprouts as an environment, (b) transcription of STEC virulence genes changed during sprouts production but generally decreased during refrigeration, and (c) STEC from fresh sprouts grown from sanitizer-treated seeds were more virulent in the animal model, but STEC from refrigerated sprouts were less virulent.

Occurrence and diversity of Shiga toxin-producing Escherichia coli (STEC) in Italian Alpine raw milk cheeses and their development in the earlier stages of different cheese-making processes

Shiga toxin-producing Escherichia coli (STEC) are foodborne pathogens implicated in diseases including hemorrhagic colitis and hemolytic uremic syndrome. In the last years, an increasing number of STEC infections associated with the consumption of raw milk cheeses have been reported, contributing to raise the public awareness. The aim of this study was to assess the occurrence of STEC biotypes in raw milk cheeses from Italian Alpine region and evaluate the influence of different scalding temperatures on their development. Six out 82 (7.3%) cheeses led to the isolation of E. coli biotypes with different virulence traits (stx1, stx2 and eae genes). To evaluate the impact of scalding temperatures on STEC growth, three E. coli biotypes were spiked into milk (<50 CFU/mL) according to the FAO and WHO recommendation. An increase in STEC counts of 3 log occurred in uncooked and semi-cooked cheeses (scalding temperature of 38 and 45 °C) while in the cooked-cheeses (54 and 56 °C) E. coli content does not exceed the 2.5 log10 CFU/g. These findings showed that raw milk cheese safety is strictly related to their production technology, highlighting the importance of the control measures at farm and dairy level to preserve the safety of these products.

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.

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.

Shiga toxin-producing Escherichia coli infection as a precipitating factor for atypical hemolytic-uremic syndrome.

Hemolytic uremic syndrome (HUS) is a thrombotic microangiopathy characterized by intravascular hemolysis. It can be classified as either typical, primarily caused by Shiga toxin-producing Escherichia coli (STEC) infection, or as atypical HUS (aHUS), which results from uncontrolled complement activation. We report the case of a 9-year-old boy with aHUS due to compound heterozygous complement factor H-related genes (CFHR) 1/3 and CFHR1-CFHR4 deletions, leading to the development of anti-complement factor H (CFH) autoantibodies. The patient presented nephrological and neurological thrombotic microangiopathy with STEC positivity. Additionally, we provide an extensive literature review of aHUS cases initially classified as typical. A total of 11 patients were included, 73% of whom were pediatric. Kidney replacement therapy was required in 73% of patients. The recurrence rate was 55%. All cases were found positive for pathological variants of the complement system genes. The most commonly implicated gene was CFH, while the CFHR genes were involved in 36% of cases, although none exhibited anti-CFH autoantibodies. Anti-complement therapy was administered in 54% of cases, and none of the patients who received it early progressed to kidney failure. STEC infection does not exclude aHUS diagnosis, and early use of anti-complement therapy might be reasonable in life-threatening conditions. Genetic testing can be helpful in patients with atypical presentations and can confirm the necessity of prolonged anti-complement therapy.

Detection of Shiga toxin-producing Escherichia coli in dairy cows: genetic characterization and inhibition of adherence by cattle anti-STEC antibodies to HEp-2 cell.

Shiga toxin-producing Escherichia coli (STEC) is a zoonotic pathogen associated with severe disease. Cattle are recognized as the primary animal reservoir of STEC. This study reports the occurrence and characterization of STEC from dairy cows and evaluates the inhibition of adherence by cattle anti-STEC antibodies to the HEp-2 cell. From 151 samples, 30% (n = 45) were positive for stx by PCR screening (25.21% of dairy cows and 46.8% of growing calves). From these positive samples, 17 STEC isolates were characterized. In dairy cows, 3 out of 17 samples carried stx2, 3 out of 17 possessed stx1, and 2 out of 17 carried stx1/stx2. In growing cows, 8 out of 17 samples carried stx1 and 1 out of 17 carried stx1/stx2. Other virulence factors such as ehxA, saa, iha, cah, and eae were detected. The strains were typed into 3 E. coli O groups (O26, O91, and O130). The analysis of the HEp-2 adherence assays indicated that all serum from both cattle categories presented high levels of inhibition of adherence. Considering the severity of the symptoms caused by STEC in humans, searching for factors influencing the colonization of STEC in cattle would help identify strategies to avoid transmission and STEC infection.

Ruminant-dense environments increase risk of Shiga toxin-producing Escherichia coli independently of ruminant contact.

Cattle and other domestic ruminants are the primary reservoirs of O157 and non-O157 Shiga toxin-producing Escherichia coli (STEC). Living in areas with high ruminant density has been associated with excess risk of infection, which could be due to both direct ruminant contact and residual environmental risk, but the role of each is unclear. We investigated whether there is any meaningful risk to individuals living in ruminant-dense areas if they do not have direct contact with ruminants. Using a Bayesian spatial framework, we investigated the association between the density of ruminants on feedlots and STEC incidence in Minnesota from 2010 to 2019, stratified by serogroup and season, and adjusting for direct ruminant contact. For every additional head of cattle or sheep per 10 acres, the incidence of O157 STEC infection increased by 30% (IRR 1.30; 95% CrI 1.18, 1.42) or 135% (IRR 2.35; 95% CrI 1.14, 4.20), respectively, during the summer months. Sheep density was also associated with O157 STEC risk during winter (IRR 4.28; 95% CrI 1.40, 8.92). The risk of non-O157 STEC infection was only elevated in areas with goat operations during summer (IRR 19.6; 95% CrI 1.69, 78.8). STEC risk associated with ruminant density was independent of direct ruminant contact across serogroups and seasons. Our findings demonstrate that living in a ruminant-dense area increases an individual's risk of O157 and non-O157 STEC infection even without direct ruminant contact, indicating that prevention efforts need to extend to community strategies for averting indirect transmission from local ruminant populations.

The impact of climate variation on the spread of Shiga toxin-producing E. coli.

Shiga toxin-producing Escherichia coli (STEC) is influenced by seasonality, but there is limited understanding of how specific climatic variables contribute to disease spread. This information aids in understanding disease transmission dynamics and could potentially inform public health modeling. This retrospective cohort study analyzed public health data from Ontario, Canada, between 2012 and 2021, along with historical climate data from Environment Canada. We employed Seasonal Autoregressive Integrated Moving Average (S-ARIMA) models to assess how temperature and precipitation impact the incidence of STEC infections, measured per 10,000,000 population. The study included 1658 confirmed STEC cases. A significant correlation was found between STEC incidence and climatic variables. Each degree Celsius increase in maximum temperature was associated with a rise of 3 STEC cases per 10,000,000 population (Centers for Disease Control and Prevention (2024)). Additionally, each millimeter of increased precipitation correlated with an increase of 1.1 cases per 10,000,000 population. The findings demonstrate a significant impact of temperature and precipitation on STEC transmission, highlighting the importance of integrating meteorological data into public health surveillance. This integration may help inform public health responses and support healthcare systems in planning for future outbreaks. Further studies are needed to refine predictive models and develop effective early warning systems for clinical settings.

Fragment Screening to Identify Inhibitors Targeting Ribosome Binding of Shiga Toxin 2.

Shiga toxins are the main virulence factors of Shiga toxin producing E. coli (STEC) and S. dysenteriae. There is no effective therapy to counter the disease caused by these toxins. The A1 subunits of Shiga toxins bind the C-termini of ribosomal P-stalk proteins to depurinate the sarcin/ricin loop. The ribosome binding site of Shiga toxin 2 has not been targeted by small molecules. We screened a fragment library against the A1 subunit of Shiga toxin 2 (Stx2A1) and identified a fragment, BTB13086, which bound at the ribosome binding site and mimicked the binding mode of the P-stalk proteins. We synthesized analogs of BTB13086 and identified a series of molecules with similar affinity and inhibitory activity. These are the first compounds that bind at the ribosome binding site of Stx2A1 and inhibit activity. These compounds hold great promise for further inhibitor development against STEC infection.

Humoral immune response to shiga toxin 2 (Stx2) in children with escherichiosis with hemolytic-uremic syndrome

Shiga toxin-producing Escherichia coli (STEC) causes acute intestinal infections and also causes acute renal failure, especially in children. Shiga toxins (Stx) occupy a central place in the pathogenesis of hemolytic uremic syndrome (HUS) in Escherichiosis. The presented work analyzes the effectiveness of laboratory diagnostics of enterohemorrhagic escherichiosis in patients at the stage of manifestation of HUS and/or acute renal failure using microbiological, immunological research methods and PCR analysis. The study used clinical material from 30 patients in the pediatric intensive care unit of the St. Vladimir Children’s City Clinical Hospital in Moscow with symptoms of HUS aged from 8 months to 5 years. Blood sera from 20 healthy donors were used as control. As a result of PCR analysis, stx2 DNA was detected in 23.3% of cases. Bacteriological research made it possible to sow a pure culture of Escherichia coli O157:H7 in only 3.3% of cases. Since the development of HUS begins in patients with acute intestinal infection caused by Shiga toxin-producing microorganisms starting from the 5th day of the disease, when antibiotic therapy is already carried out, the bacteria can be completely destroyed, which makes it difficult to identify them by bacteriological methods, as well as to detect genes encoding Shiga toxin in PCR analysis. Typically, patients with HUS are admitted to the intensive care unit 5–7 days after the onset of the disease, when class G immunoglobulins specific to the pathogen are already beginning to circulate in the blood. In this regard, the use of immunological tests can be effective to confirm the diagnosis of STEC infection. In our studies, enzyme immunoassay allowed us to detect antibodies to Stx2A in 63.3% and to Stx2B in 43.3% of patients. Using immunoblotting, antibodies to Stx2A were detected in all sera obtained from patients and in 66.7% of cases to Stx2B. Immunoblot analysis was characterized by higher sensitivity for detecting antibodies to Stx2, however, due to the presence of an immunological layer among healthy people, it is preferable to use ELISA analysis. In healthy donors with antibodies to Stx2, the antibody titer was significantly lower than in patients. Laboratory confirmation of the diagnosis of STEC infection is difficult when conducting microbiological and molecular genetic studies, which is confirmed in this work. The effectiveness of laboratory diagnostics can be expanded by performing an ELISA aimed at detecting antibodies to Stx2A.

Shiga Toxin-Producing Escherichia coli Strains from Romania: A Whole Genome-Based Description.

The zoonotic Shiga toxin-producing Escherichia coli (STEC) group is unanimously regarded as exceptionally hazardous for humans. This study aimed to provide a genomic perspective on the STEC recovered sporadically from humans and have a foundation of internationally comparable data. Fifty clinical STEC isolates, representing the culture-confirmed infections reported by the STEC Reference Laboratory between 2016 and 2023, were subjected to whole-genome sequencing (WGS) analysis and sequences were interpreted using both commercial and public free bioinformatics tools. The WGS analysis revealed a genetically diverse population of STEC dominated by non-O157 serogroups commonly reported in human STEC infections in the European Union. The O26:H11 strains of ST21 lineage played a major role in the clinical disease resulting in hospitalisation and cases of paediatric HUS in Romania surpassing the O157:H7 strains. The latter were all clade 7 and mostly ST1804. Notably, among the Romanian isolates was a stx2a-harbouring cryptic clade I strain associated with a HUS case, stx2f- and stx2e-positive strains, and hybrid strains displaying a mixture of intestinal and extraintestinal virulence genes were found. As a clearer picture emerges of the STEC strains responsible for infections in Romania, further surveillance efforts are needed to uncover their prevalence, sources, and reservoirs.

Vacation in Egypt associated with Shiga toxin-producing Escherichia coli infection in children and adolescents, northern Italy, 2023.

BackgroundHaemolytic uremic syndrome (HUS) is a severe complication of infection with Shiga toxin-producing Escherichia coli (STEC). Although the reservoirs of STEC are known, the source of the infection of sporadic cases is often unknown. In 2023, we observed several cases of bloody diarrhoea with STEC infection in children and adolescents returning from vacations.AimWe aimed to explore the association between travel and bloody diarrhoea with STEC infection in children and adolescents.MethodsWe included all children and adolescents with bloody diarrhoea with STEC infection identified in 2023 by the ItalKid-HUS Network surveillance system in northern Italy. We interviewed children's families and sent a questionnaire on recent travels abroad. The exposure time was between 3 days after arrival abroad and 5 days after return home. A self-controlled case series (SCCS) design was used in the analysis.ResultsOf the 43 cases, 11 developed HUS. Twenty-three cases did not travel abroad, while 20 had travelled to several destinations. The incidence rate ratio (IRR) associated with travel to Egypt was 88.6 (95% confidence interval (CI): 17.0-462). Serotype analysis excluded the possibility of a single strain causing the infections. We did not find the source of the infections.ConclusionThere is an elevated risk of acquiring STEC infection with bloody diarrhoea and HUS associated with travel to Egypt. Specific investigations to identify the source are needed to implement effective preventive measures.

How to Improve Surveillance Program for Shiga Toxin-Producing E. coli (STEC): Gap Analysis and Pilot Study.

Several pathotypes of enteric E. coli have been identified. The group represented by Shiga toxin-producing E. coli (STEC) is of particular interest. Raw milk and raw milk products are significant sources of STEC infection in humans; therefore, identifying pathogens at the herd level is crucial for public health. Most national surveillance programs focus solely on raw milk and raw milk cheeses that are ready for retail sale, neglecting the possibility of evaluating the source of contamination directly at the beginning of the dairy chain. To assess the viability of the application of new molecular methodologies to STEC identification in raw milk filters and in calf feces, we analyzed 290 samples from 18 different dairy herds, including 88 bulk tank milk (BTM), 104 raw milk filters (RMF), and 98 calf feces samples. In total 3.4% of BTM, 41.4% of RMF, and 73.4% of calves' feces were positive for stx, supporting our hypothesis that BTM is not a suitable matrix to assess the presence of STEC at herd level, underestimating it. Our conclusion is that the surveillance program needs critical and extensive improvements such as RMF and calves' feces analysis implementation to be more efficient in detecting and preventing STEC infections. The epidemiology of these infections and the characteristics of the pathogen clearly show how a One Health approach will be pivotal in improving our capabilities to control the spread of these infections.