Shiga Toxin-producing Escherichia Coli O157 Research Articles

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.

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.

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.

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.

79 Microbiological effects of control measures during pork production

Abstract The objective of this study was to determine the prevalence of Shiga toxin-producing Escherichia coli (STEC) O157:H7 and efficacy of control measures at a pork processing facility. Outbreaks of E. coli O157:H7 associated with pork, though not common globally, have been reported multiple times in recent years in Alberta. Sampling at a commercial facility processing hogs was carried out at monthly intervals for 11 mo. At the facility, the dressed carcasses are sprayed with 5% lactic acid (LA) and immediately air-chilled after the LA spray. At each sampling time, 14 different types of samples were collected: dirt from the holding pen floor (n = 2), gloves of workers (n = 5), scalding water (n = 1), tonsils (n = 5), cecal material (n = 5), five swabs each from hides of animals after bleeding (HAB), and carcasses after scalding (CAS), before evisceration (CBE), after evisceration (CAE), before LA spray (BLA), after LA spray (ALA), 1h after LA spray (ALA1), 2h after LA spray (ALA2), and after chilling (ACH). All samples were enumerated for total plate count (TPC), Enterobacteriaceae, and E. coli using specific media for each. In addition, an aliquot of each sample was enriched and tested for the presence of the O-antigen gene (rfbE) for E. coli O157 and Shiga toxin-producing gene (stx1/stx2) using real-time PCR. Cultures positive for both rfbE and stx were further plated for the isolation of E. coli O157:H7. Of the 683 enrichment cultures, 12 were positive for both marker genes, from cecal material (n = 2), dirt (n = 4) and HAB (n = 6). However, none of these samples yielded STEC O157 isolates. Of these enrichment cultures, 69, 45 and 32 were positive for stx1, stx2, and both stx genes, respectively. Positive samples were mostly associated with HAB, cecal material and dirt. HAB was most and least contaminated in August/June and December, differing by 1 log unit for TPC (P < 0.05). Enterobacteriaceae and E. coli on HAB were also affected by month of sampling (P < 0.05), but this temporal trend differed from TPC (Table). The numbers for TPC, Enterobacteriaceae, and E. coli all decreased (P < 0.05) as the stage of carcass processing progressed. The overall reduction of TPC and E. coli was 7 and 5 log units, respectively. Scalding, LA spray and air chilling all reduced (P < 0.05) the numbers of indicator bacteria on carcasses. Of the 55 samples each collected from BLA, ALA, ALA1, ALA2, and ACH, 36, 13, 2, 1 and 0 were positive for E. coli, respectively. In conclusion, pork processing environments can be contaminated with STEC from hides, feces and holding pen, but levels of STEC O157 appear to be low. Scalding, LA spray and air chilling can be effective measures to reduce the risk of bacterial contamination of pork.

A review of conditions influencing fate of Shiga toxin-producing Escherichia coli O157:H7 in leafy greens.

The accuracy of predictive microbial models used in quantitative microbial risk assessment (QMRA) relies on the relevancy of conditions influencing growth or inactivation. The continued use of log-linear models in studies remains widespread, despite evidence that they fail to accurately account for biphasic kinetics or include parameters to account for the effect of environmental conditions within the model equation. Although many experimental studies detail conditions of interest, studies that do not do so lead to uncertainty in QMRA modeling because the applicability of the predictive microbial models to the conditions in the risk scenarios is questionable or must be extrapolated. The current study systematically reviewed 65 articles that provided quantitative data and documented the conditions influencing the inactivation or growth of Shiga toxin-producing Escherichia coli (STEC) O157:H7 in leafy greens. The conditions were identified and categorized as environmental, biological, chemical, and/or processing. Our study found that temperature (n=37 studies) and sanitizing and washing procedures (n=12 studies) were the most studied conditions in the farm-to-table continuum of leafy greens. In addition, relative humidity was also established to affect growth and inactivation in more than one stage in the continuum. This study proposes the evaluation of the interactive effects of multiple conditions in processing and storage stages from controlled experiments as they relate to the fate of STEC O157:H7 in leafy greens for future quantitative analysis.

A Health Threat from Farm to Fork: Shiga Toxin-Producing Escherichia coli Co-Harboring blaNDM-1 and mcr-1 in Various Sources of the Food Supply Chain.

The dissemination of resistant pathogens through food supply chains poses a significant public health risk, spanning from farm to fork. This study analyzed the distribution of Shiga toxin-producing Escherichia coli (STEC) across various sources within the animal-based food supply chain. A total of 500 samples were collected from livestock, poultry, the environment, fisheries, and dairy. Standard microbiological procedures were employed to isolate and identify E. coli isolates, which were further confirmed using MALDI-TOF and virulence-associated genes (VAGs) such as stx1, stx2, ompT, hylF, iutA, fimH, and iss. The phenotypic resistance patterns of the isolates were determined using the disc diffusion method, followed by molecular identification of antibiotic resistance genes (ARGs) through PCR. STEC were subjected to PCR-based O typing using specific primers for different O types. Overall, 154 (30.5%) samples were confirmed as E. coli, of which 77 (50%) were multidrug-resistant (MDR) E. coli. Among these, 52 (67.53%) isolates exhibited an array of VAGs, and 21 (40.38%) were confirmed as STEC based on the presence of stx1 and stx2. Additionally, 12 out of 52 (23.07%) isolates were identified as non-O157 STEC co-harbouring mcr-1 and blaNDM-1. O26 STEC was found to be the most prevalent among the non-O157 types. The results suggest that the detection of STEC in food supply chains may lead to serious health consequences, particularly in developing countries with limited healthcare resources.

Prevalence, Molecular Characterization, and Antimicrobial Resistance Profiles of Shiga Toxin-Producing Escherichia coli Isolated from Raw Beef, Pork, and Chicken Meat in Vietnam.

Shiga toxin-producing Escherichia coli (STEC) is one of the most important foodborne pathogens, and the rise of antibiotic resistance to it is a significant threat to global public health. The purpose of this study is to investigate the prevalence, molecular characterization, and antibiotic resistance of STEC isolated from raw meat in Vietnam. The findings in this study showed that the prevalence of STEC in raw beef, pork, and chicken meat was 9.72% (7/72), 5.56% (4/72), and 1.39% (1/72), respectively. The STEC isolates were highly resistant to ampicillin (91.67%) and tetracycline (91.67%), followed by trimethoprim/sulfamethoxazole (83.33%), streptomycin (75%), and florfenicol (66.67%). The incidence of STEC virulence-associated genes, including stx1, stx2, eae, and ehxA, was 8.33% (1/12), 91.67% (11/12), 33.33% (4/12), and 58.33% (7/12), respectively. STEC serogroups O157, O26, and O111 were detected in 3 out of 12 STEC isolates. Two isolates were found to be ESBL producers carrying the blaCTX-M-55 gene, and three isolates were colistin-resistant strains harboring the mcr-1 gene. Notably, a STEC O111 isolate from chicken meat harbored both the blaCTX-M-55 and mcr-1 genes.

Shiga toxin-producing Escherichia coli O157:H7 outbreak associated with school field trips at a farm animal exhibit-Tennessee, September-October 2023.

In October 2023, the Tennessee Department of Health identified an outbreak of Shiga toxin-producing Escherichia coli (STEC) O157:H7 infections among elementary school students who attended school field trips to the same farm animal exhibit. Our aim was to determine STEC source and prevent additional illnesses by initiating epidemiologic, laboratory and environmental investigations. We identified cases using laboratory-based surveillance and by surveying caregivers of children who attended the exhibit. Probable cases were defined as illness with abdominal cramps or diarrhoea after attendance; confirmed cases were laboratory-confirmed STEC infection in an attendee or household contact. A site visit was conducted, and event organizers were interviewed. Human stool, animal faeces and environmental samples were tested for STEC O157:H7 by real-time polymerase chain reaction (PCR), culture and whole-genome sequencing (WGS). Approximately 2300 elementary school students attended the animal exhibit during 2 days. Field trip activities included contact with different farm animal species, drinking pasteurized milk outside animal enclosures and eating lunch in a separate building onsite. We received survey responses from 399 caregivers for 443 (19%) animal exhibit attendees. We identified 9 confirmed and 55 probable cases with illness onset dates during 26 September to 12 October. Seven children aged 1-7 years were hospitalized. Four children aged 1-6 years experienced haemolytic uraemic syndrome; none died. Laboratory testing identified STEC O157:H7 by culture from eight human stool samples with 0-1 allele difference by WGS. Three environmental samples had Shiga toxin (stx 2) genes detected by PCR, but no STEC isolates were recovered by culture. This is the largest reported STEC O157:H7 outbreak associated with an animal exhibit in Tennessee. We identified opportunities for educating school staff, event organizers and families about zoonotic disease risks associated with animal contact and published prevention measures.

Increased resistance against tellurite is conferred by a mutation in the promoter region of uncommon tellurite resistance gene tehB in the ter-negative Shiga toxin-producing Escherichia coli O157:H7.

Selective agents are important for isolating Shiga toxin-producing Escherichia coli (STEC) because the undesirable growth of microflora should be inhibited. Potassium tellurite (PT) is a common selective agent for major STEC serotypes. In this study, we found a novel variant of PT resistance genes, tehAB, in STEC O157:H7. Molecular experiments clearly showed that one point mutation in a predicted internal promoter region of tehB upregulated the expression of the gene and consequently led to increased resistance to PT. Because tehAB genes are ubiquitous across E. coli, these results provide universal insight into PT resistance in this species.

Foodborne Illnesses from Leafy Greens in the United States: Attribution, Burden, and Cost

Leafy green vegetables are a major source of foodborne illnesses. Nevertheless, few studies have attempted to estimate attribution and burden of illness estimates for leafy greens. This study combines results from three outbreak-based attribution models with illness incidence and economic cost models to develop comprehensive pathogen-specific burden estimates for leafy greens and their subcategories in the United States. We find that up to 9.18% (90% CI: 5.81%-15.18%) of foodborne illnesses linked to identified pathogens are attributed to leafy greens. Including ‘Unknown’ illnesses not linked to specific pathogens, leafy greens account for as many as 2,307,558 (90% CI: 1,077,815–4,075,642) illnesses annually in the United States. The economic cost of these illnesses is estimated to be up to $5.278 billion (90% CI: $3.230-$8.221 billion) annually. Excluding the pathogens with small outbreak sizes, Norovirus, Shiga toxin-producingEscherichia coli (both non-O157 and O157:H7), Campylobacter spp., and nontyphoidal Salmonella, are associated with the highest number of illnesses and greatest costs from leafy greens. While lettuce (romaine, iceberg, “other lettuce”) takes 60.8% of leafy green outbreaks, it accounts for up to 75.7% of leafy green foodborne illnesses and 70% of costs. Finally, we highlighted that 19.8% of Shiga toxin-producingEscherichia coli O157:H7 illnesses are associated with romaine among all food commodities, resulting in 12,496 estimated illnesses and $324.64 million annually in the United States.

Survival of O157 and non-O157 shiga toxin-producing Escherichia coli in Korean style kimchi

Korean style kimchi contaminated with Shiga toxin-producing Escherichia coli (STEC) O157:H7 was the cause of an outbreak in Canada from December 2021 to January 2022. To determine if this STEC O157:H7 has greater potential for survival in kimchi than other STEC, the outbreak strain and six other STEC strains (O26:H11, O91:H21, O103:H2, O121:H19, and two O157:H7) were inoculated individually at 6 to 6.5 log CFU/g into commercially sourced kimchi and incubation at 4 °C. At intervals of seven days inoculated and control kimchi was plated onto MacConkey agar to enumerate lactose utilising bacteria. The colony counts were interpreted as enumerating the inoculated STEC, since no colonies were observed on MacConkey agar plated with uninoculated kimchi. Over eight weeks of incubation the pH was stable at 4.10 to 4.05 and the STEC strains declined by 0.7–1.0 log, with a median reduction of 0.9 log. The linear rate of reduction of kimchi outbreak STEC O157:H7 was −0.4 log per 30 days (Slope Uncertainty 0.05), which was not significantly different from the other O157 and nonO157 STEC strains (P = 0.091). These results indicate that the outbreak was not due to the presence of strain better adapted to survival in kimchi than other STEC, and that STEC can persist in refrigerated Korean style kimchi with a minimal decline over the shelf-life of the product.

Isolation of Shiga Toxin-Producing Escherichia coli from the Surfaces of Beef Carcasses in Slaughterhouses in Japan

Shiga toxin-producing E. coli (STEC) is an important foodborne pathogen worldwide. It is necessary to control and prevent STEC contamination on beef carcasses in slaughterhouses because STEC infection is associated with beef consumption. However, the frequencies of STEC contamination of beef carcasses in various slaughterhouses in Japan are not well known. Herein, we investigated the contamination of beef carcasses with STEC in slaughterhouses to assess the potential risks of STEC. In total, 524 gauze samples were collected from the surfaces of beef carcasses at 12 domestic slaughterhouses from November 2020 to February 2023. The samples were measured for aerobic plate counts and tested for pathogenic genes (stx and eae) and major O-serogroups (O26, O45, O103, O111, O121, O145, and O157) by real-time PCR screening. Subsequently, immunomagnetic separation (IMS) was performed on samples positive for stx, eae, and at least one of the seven O-serogroups of STEC. Isolation process without IMS was performed on samples positive for stx, including those subjected to IMS. STEC O157:H7 and stx-positive E. coli other than serotype O157:H7 were isolated from 0.6% and 4.6% of beef carcass surfaces, respectively. Although the STEC O157:H7 isolation rate was low and stx-positive E. coli other than serotype O157:H7 belonged to minor O-serogroups, the results mean a risk of foodborne illness. Furthermore, a moderate correlation was observed between aerobic plate counts and detection rates of stx-positive samples by real-time PCR screening. The STEC O157:H7 isolated facilities showed higher values on aerobic plate counts and detection rates of stx-positive samples than the mean values of total samples. Therefore, these results suggest that it is important to evaluate hygiene treatments against beef carcasses for the reduction of STEC contamination risk, particularly in facilities with high aerobic plate counts.

Virulence and Antimicrobial Resistance Profiles of Shiga Toxin-Producing Escherichia coli from River Water and Farm Animal Feces near an Agricultural Region in Northwestern Mexico

Shiga toxin-producing Escherichia coli (STEC) are zoonotic enteric pathogens linked to human gastroenteritis worldwide. To aid the development of pathogen control efforts, the present study characterized the genotypic diversity and pathogenic potential of STEC recovered from sources near agricultural fields in Northwest Mexico. Samples were collected from irrigation river water and domestic animal feces in farms proximal to agricultural fields and were subjected to enrichment followed by immunomagnetic separation and plating on selective media for the recovery of the STEC isolates. Comparative genomic analyses indicated that the recovered STEC with the clinically relevant serotypes O157:H7, O8:H19, and O113:H21 had virulence genes repertoires associated with host cell adherence, iron uptake and effector protein secretion. Subsequent phenotypic characterization revealed multidrug resistance against aminoglycoside, carbapenem, cephalosporin, fluoroquinolone, penicillin, phenicol, and tetracycline, highlighting the need for improved surveillance on the use of antimicrobials. The present study indicated for the first time that river water in the agricultural Culiacan Valley in Mexico is a relevant key route of transmission for STEC O157 and non-O157 with a virulence potential. In addition, feces from domestic farm animals near surface waterways can act as potential point sources of contamination and transport of diverse STEC with clinically relevant genotypes.

Transcription levels of hes and their involvement in the biofilm formation of Shiga toxin-producing Escherichia coli O91.

Shiga toxin-producing Escherichia coli (STEC) are recognized as being responsible for many cases of foodborne diseases worldwide. Cattle are the main reservoir of STEC, shedding the microorganisms in their feces. The serogroup STEC O91 has been associated with hemorrhagic colitis and hemolytic uremic syndrome. Locus of Adhesion and Autoaggregation (LAA) and its hes gene are related to the pathogenicity of STEC and the ability to form biofilms. Considering the frequent isolation of STEC O91, the biofilm-forming ability, and the possible role of hes in the pathogenicity of STEC, we propose to evaluate the ability of STEC to form biofilms and to evaluate the expression of hes before and after of biofilm formation. All strains were classified as strong biofilm-forming. The hes expression showed variability between strains before and after biofilm formation, and this may be due to other genes carried by each strain. This study is the first to report the relationship between biofilm formation, and hes expression and proposes that the analysis and diagnosis of LAA, especially hes as STEC O91 virulence factors, could elucidate these unknown mechanisms. Considering that there is no specific treatment for HUS, only supportive care, it is necessary to know the survival and virulence mechanisms of STEC O91.

Linking active rectal mucosa-attached microbiota to host immunity reveals its role in host-pathogenic STEC O157 interactions.

The rectal-anal junction (RAJ) is the major colonization site of Shiga toxin-producing Escherichia coli (STEC) O157 in beef cattle, leading to transmission of this foodborne pathogen from farms to food chains. To date, there is limited understanding regarding whether the mucosa-attached microbiome has a profound impact on host-STEC interactions. In this study, the active RAJ mucosa-attached microbiota and its potential role in host immunity-STEC commensal interactions were investigated using RAJ mucosal biopsies collected from calves orally challenged with two STEC O157 strains with or without functional stx2a (stx2a+ or stx2a-). The results revealed that shifts of microbial diversity, topology, and assembly patterns were subjected to stx2a production post-challenge and Paeniclostridium and Gallibacterium were the keystone taxa for both microbial interactions and assembly. Additional mucosal transcriptome profiling showed stx2a-dependent host immune responses (i.e. B- and T-cell signaling and antigen processing and presentation) post-challenge. Further integrated analysis revealed that mucosa-attached beneficial microbes (i.e. Provotella, Faecalibacterium, and Dorea) interacted with host immune genes pre-challenge to maintain host homeostasis; however, opportunistic pathogenic microbes (i.e. Paeniclostridium) could interact with host immune genes after the STEC O157 colonization and interactions were stx2a-dependent. Furthermore, predicted bacterial functions involved in pathogen (O157 and Paeniclostridium) colonization and metabolism were related to host immunity. These findings suggest that during pathogen colonization, host-microbe interactions could shift from beneficial to opportunistic pathogenic bacteria driven and be dependent on the production of particular virulence factors, highlighting the potential regulatory role of mucosa-attached microbiota in affecting pathogen-commensal host interactions in calves with STEC O157 infection.

Shedding and exclusion from childcare in children with Shiga toxin-producing Escherichia coli, 2018-2022.

Excluding children with Shiga toxin-producing Escherichia coli (STEC) from childcare until microbiologically clear of the pathogen, disrupts families, education, and earnings. Since PCR introduction, non-O157 STEC serotype detections in England have increased. We examined shedding duration by serotype and transmission risk, to guide exclusion advice. We investigated STEC cases aged <6years, residing in England and attending childcare, with diarrhoea onset or sample date from 31 March 2018 to 30 March 2022. Duration of shedding was the interval between date of onset or date first positive specimen and earliest available negative specimen date. Transmission risk was estimated from proportions with secondary cases in settings attended by infectious cases. There were 367 cases (STEC O157 n=243, 66.2%; STEC non-O157 n=124, 33.8%). Median shedding duration was 32days (IQR 20-44) with no significant difference between O157 and non-O157; 2% (n=6) of cases shed for ≥100days. Duration of shedding was reduced by 17% (95% CI 4-29) among cases reporting bloody diarrhoea. Sixteen settings underwent screening; four had secondary cases (close contacts' secondary transmission rate=13%). Shedding duration estimates were consistent with previous studies (median 31days, IQR 17-41). Findings do not warrant guidance changes regarding exclusion and supervised return of prolonged shedders, despite serotype changes.

Escherichia coli O157:H7 tir 255 T > A allele strains differ in chromosomal and plasmid composition.

Shiga toxin-producing Escherichia coli (STEC) O157:H7 strains with the T allele in the translocated intimin receptor polymorphism (tir) 255 A > T gene associate with human disease more than strains with an A allele; however, the allele is not thought to be the direct cause of this difference. We sequenced a diverse set of STEC O157:H7 strains (26% A allele, 74% T allele) to identify linked differences that might underlie disease association. The average chromosome and pO157 plasmid size and gene content were significantly greater within the tir 255 A allele strains. Eighteen coding sequences were unique to tir 255 A allele chromosomes, and three were unique to tir 255 T allele chromosomes. There also were non-pO157 plasmids that were unique to each tir 255 allele variant. The overall average number of prophages did not differ between tir 255 allele strains; however, there were different types between the strains. Genomic and mobile element variation linked to the tir 255 polymorphism may account for the increased frequency of the T allele isolates in human disease.

Effect of electron beam irradiation on minas frescal cheese artificially contaminated with Shiga toxin-producing Escherichia coli O157:H7

Minas frescal cheese (MFC) is a potential vehicle for Shiga toxin-producing Escherichia coli (STEC) O157:H7. The effect of electron beam irradiation on the control of STEC O157:H7 in artificially contaminated MFC (5 log cfu g−1) was evaluated. Samples were irradiated at doses of 1.0, 1.5, and 2.0 kGy and evaluated for 40 days at 4 °C. Shelf life extension and the stability of the lipid fraction of the cheeses were also evaluated. At a dose of 1.0 kGy, STEC O157:H7 was reduced by 3 log cfu g−1, while at 1.5 and 2.0 kGy, STEC O157:H7 was eliminated. Shelf life extension, measured by the count of total aerobic bacteria (TAB), occurred in up to ten days. Irradiated and non-irradiated samples were affected by lipid oxidation. Electron beam irradiation proved to be a viable technology and a good alternative to guarantee the quality and control of STEC O157:H7 in the MFC.

Prevalence, characterization and antimicrobial resistance of non-O157 Shiga toxin-producing Escherichia coli isolates from healthy cattle in Tunisia

AbstractAs a continuation of our previous work, here we examined the occurrence, virulence factors and antibiotic resistance of non-O157 Shiga toxin-producing Escherichia coli (STEC) bacteria in faecal samples of healthy cattle in Tunisia. From 260 faecal samples, 236 E. coli isolates were obtained, of which 30 (12.7%) proved to be non-O157:H7 STEC. The presence of STEC-serogroup virulence genes stx1, stx2, eaeA and ehxA was detected by PCR. Further virulence genes and other genes used for phylogenetic grouping were also examined among the STEC isolates. The most dominant serogroup was O145, followed by O103 and O45. Shiga toxin gene (stx1) was found in 70% of the STEC strains, stx2 and ehxA were detected in 17 isolates (56.7%). Other virulence genes, including fimH, traT, sfa/focDE, iutA, cdt3, hly, fyuA and ibeA, were detected in 100%, 60%, 46.7%, 46.7%, 36.7%, 10%, 6.7% and 3.3%, of the strains, respectively. More than 70% of the isolates were susceptible to most of the tested antibiotics. We concluded that the presence of STEC O145, O103 and O45 (12.7%), together with the earlier described STEC O157:H7 (4.2%) in cattle, intended for slaughter, may represent a threat to public health. Efficient control measures are needed in the farms and slaughterhouses in Tunisia.