A COHORT STUDY ON SHIGA TOXIN PRODUCING E. COLI O157:H7 ISOLATED FROM SOME MEAT PRODUCTS IN ASSIUT GOVERNORATE AS A CAUSE OF BLOODY DIARRHEA IN CHILDREN

Abstract Background: Shiga toxin-producing Escherichia coli (STEC) are associated with major food illness around the world. E.coli O157, has been widely reported as the most common STEC serogroup, and has emerged as an important enteric pathogen. Further, cattle have been identified as a major E. coli O157:H7 reservoir for human infection; however, the ecology of this organism in camels, sheep and goats is less understood. The current study aims to evaluate the prevalence of E. coli serotype O157 in feces of cattle, camels, sheep and goats slaughtered in United Arab Emirates (UAE) for meat consumption. This study was carried out on fecal samples of healthy cattle (n = 137), camels (n = 140), sheep (n = 141) and goats (n = 150) during the period of September 2017 to August 2018. We have used the traditional sensitive immunomagnetic separation technique (IMS) coupled with a culture plating method for detection of E. coli O157. Non-sorbitol fermenting colonies were assessed via the latex agglutination test and the positive cultures were subjected to PCR for detection of attaching and effacing genes (eaeA), hemolysin A (hlyA) and Shiga toxin-producing genes (stx1 and stx2) and genes specific for E. coli O157:H7 (rfb O157, uid A and flic H7). All E. coli O157 isolates were analyzed for their susceptibility pattern toward 20 select antibiotics.Results: E. coli O157 was present in the fecal samples of goats, camels and cattle at 2%, 3.3%, and 1.6%, respectively. In sheep we failed to detect any E. coli O157 strains. The most prevalent E.coli O157 gene identified across all species’ isolates was stx2, while stx1 was not detected in any of the samples. After testing samples from camels, goats and cattle, Cefotaxime (100%), Chloramphenicol (100%), Ciprofloxacin (100%), Norfloxacin (100%) and Polymixin B (100%) showed susceptibility showed susceptibility to all E.coli O157 isolates.Conclusion: This is the first study, to our knowledge, to report on the prevalence of E. coli O157 in the slaughter animals in UAE and clearly demonstrates the presence of these pathogens in slaughtered animals, which could possibly contaminate the meat products intended for human consumption.

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Shiga toxin-producing Escherichia coli in meat and vegetable products in Emilia Romagna Region, years 2012-2013

Italian Journal of Food Safety ◽

10.4081/ijfs.2015.4511 ◽

2015 ◽

Vol 4 (1) ◽

Cited By ~ 4

Author(s):

Lia Bardasi ◽

Roberta Taddei ◽

Lucia Nocera ◽

Matteo Ricchi ◽

Giuseppe Merialdi

Keyword(s):

Escherichia Coli ◽

Shiga Toxin ◽

Virulence Genes ◽

Meat Products ◽

Gene Isolation ◽

E Coli ◽

Vegetable Products ◽

Emilia Romagna Region ◽

Meat Samples ◽

Pcr Targeting

In 2012-2013 Emilia-Romagna Region introduced a monitoring plan for Shiga toxin-producing Escherichia coli (STEC) in foodstuff. Six hundred eighty-nine meat samples and 273 fruit and vegetable products were analyzed according to ISOTS13136. Pre-enriched samples were tested by multiplex real time PCR targeting the virulence genes eae, stx1 and stx2. Stx2 positive samples were investigated for the presence of serogroup O104 associated gene. O103, O111, O145, O157, O26 associated genes were tested on samples positive for stx in association with eae gene. Isolation of E. coli strains was attempted from samples positive for serogroup-associated genes. Thirtyfour meat products (4.9%) resulted positive for stx1 and/or stx2 genes and 46 (6.7%) for stx1 and/or stx2 genes in association with eae gene. Forty-five (6.5%) samples resulted positive at least at one serogroup. Serogroup O103, O104, O111, O145, O157 and O26 genes were detected respectively in 1.3, 0.3, 0.1, 3.9, 2.9 and 2.5% samples; 0.6% samples resulted positive for STEC isolation (2 E. coli O103 and 2 E. coli O157). It is worth noting that STEC virulence genes were detected at high frequency (19%) in fresh pork meat sausages. Four (1.5%) vegetable samples were positive for stx1 and/or stx2 genes and 1 (0.4%) for stx1 and/or stx2 genes in association with eae gene; none resulted positive for the tested serogroups. Only a low number of samples positive by molecular methods were confirmed by cultural isolation. It is therefore of the uttermost importance for appropriate risk management, to be fully aware of the meaning of the analytical result.

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Occurrence, virulence genes, and antimicrobial profiles of Escherichia coli O157 isolated from ruminants slaughtered in Al Ain, United Arab Emirates

10.21203/rs.2.22016/v2 ◽

2020 ◽

Author(s):

Dawood Al-Ajmi ◽

Shafeeq Rahman ◽

Sharmila Banu

Keyword(s):

Escherichia Coli ◽

United Arab Emirates ◽

Shiga Toxin ◽

Meat Products ◽

Immunomagnetic Separation ◽

Latex Agglutination ◽

Human Consumption ◽

Fecal Samples ◽

Chain Reactions ◽

E Coli

Abstract Background: Shiga toxin-producing Escherichia coli (STEC) is a major source of food-borne illness around the world. E. coli O157 has been widely reported as the most common STEC serogroup and has emerged as an important enteric pathogen. Cattle, in particular have been identified as a major E. coli O157:H7 reservoir of human infections; however, the prevalence of this organism in camels, sheep, and goats is less understood. The aim of this study was to evaluate the occurrence and concentration of E. coli serotype O157 in the feces of healthy camels (n = 140), cattle (n =137), sheep (n = 141) and goats (n = 150) slaughtered in United Arab Emirates (UAE) for meat consumption between September 2017 and August 2018. We used immunomagnetic separation coupled with a culture-plating method to detect E. coli O157. Non-sorbitol fermenting colonies were assessed via latex-agglutination testing, and positive cultures were analyzed by performing polymerase chain reactions to detect genes encoding attaching and effacing protein (eaeA), hemolysin A (hlyA, also known as ehxA) and Shiga toxin (stx1 and stx2), and E. coli O157:H7 specific genes (rfb O157, uidA, and fliC). All E. coli O157 isolates were analyzed for their susceptibility to 20 selected antimicrobials.Results: E. coli O157 was observed in camels, goats, and cattle fecal samples at abundances of 4.3%, 2%, and 1.46%, respectively, but it was undetectable in sheep feces. The most prevalent E. coli O157 gene in all STEC isolates was stx2;, whereas, stx1 was not detected in any of the samples. The fecal samples from camels, goats, and cattle harbored E. coli O157 isolates that were 100% susceptible to cefotaxime, chloramphenicol, ciprofloxacin, norfloxacin, and polymyxin B.Conclusion: To our knowledge, this is the first report on the occurrence of E. coli O157 in slaughter animals in the UAE. Our results clearly demonstrate the presence of E. coli O157 in slaughtered animals, which could possibly contaminate meat products intended for human consumption.

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Validation of the Applied Biosystems RapidFinder Shiga Toxin–Producing E. coli (STEC) Detection Workflow

Journal of AOAC International ◽

10.5740/jaoacint.16-0235 ◽

2016 ◽

Vol 99 (6) ◽

pp. 1537-1554 ◽

Cited By ~ 3

Author(s):

Jonathan Cloke ◽

Sharon Matheny ◽

Michelle Swimley ◽

Robert Tebbs ◽

Angelia Burrell ◽

...

Keyword(s):

Escherichia Coli ◽

Shiga Toxin ◽

Method Comparison ◽

Meat Products ◽

Ground Beef ◽

Reference Method ◽

Acid Extraction ◽

Screening Assay ◽

Isolation And Identification ◽

E Coli

Abstract The Applied Biosystems™ RapidFinder™ STEC Detection Workflow (Thermo Fisher Scientific) is a complete protocol for the rapid qualitative detection of Escherichia coli (E. coli) O157:H7 and the “Big 6” non-O157 Shiga-like toxin-producing E. coli (STEC) serotypes (defined as serogroups: O26, O45, O103, O111, O121, and O145). The RapidFinder STEC Detection Workflow makes use of either the automated preparation of PCR-ready DNA using the Applied Biosystems PrepSEQ™ Nucleic Acid Extraction Kit in conjunction with the Applied Biosystems MagMAX™ Express 96-well magnetic particle processor or the Applied Biosystems PrepSEQ Rapid Spin kit for manual preparation of PCR-ready DNA. Two separate assays comprise the RapidFinder STEC Detection Workflow, the Applied Biosystems RapidFinder STEC Screening Assay and the Applied Biosystems RapidFinder STEC Confirmation Assay. The RapidFinder STEC Screening Assay includes primers and probes to detect the presence of stx1 (Shiga toxin 1), stx2 (Shiga toxin 2), eae (intimin), and E. coli O157 gene targets. The RapidFinder STEC Confirmation Assay includes primers and probes for the “Big 6” non-O157 STEC and E. coli O157:H7. The use of these two assays in tandem allows a user to detect accurately the presence of the “Big 6” STECs and E. coli O157:H7. The performance of the RapidFinder STEC Detection Workflow was evaluated in a method comparison study, in inclusivity and exclusivity studies, and in a robustness evaluation. The assays were compared to the U.S. Department of Agriculture (USDA), Food Safety and Inspection Service (FSIS) Microbiology Laboratory Guidebook (MLG) 5.09: Detection, Isolation and Identification of Escherichia coli O157:H7 from Meat Products and Carcass and Environmental Sponges for raw ground beef (73% lean) and USDA/FSIS-MLG 5B.05: Detection, Isolation and Identification of Escherichia coli non-O157:H7 from Meat Products and Carcass and Environmental Sponges for raw beef trim. No statistically significant differences were observed between the reference method and the individual or combined kits forming the candidate assay using either of the DNA preparation kits (manual or automated extraction). For the inclusivity and exclusivity evaluation, the RapidFinder STEC Detection Workflow, comprising both RapidFinder STEC screening and confirmation kits, correctly identified all 50 target organism isolates and correctly excluded all 30 nontarget strains for both of the assays evaluated. The results of these studies demonstrate the sensitivity and selectivity of the RapidFinder STEC Detection Workflow for the detection of E. coli O157:H7 and the “Big 6” STEC serotypes in both raw ground beef and beef trim. The robustness testing demonstrated that minor variations in the method parameters did not impact the accuracy of the assay and highlighted the importance of following the correct incubation temperatures

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Molecular characterization and diversity of shiga toxin-producing E. coli (STEC) isolates obtained in the Netherlands

10.26226/morressier.56d5ba26d462b80296c94bac ◽

2016 ◽

Author(s):

Mithila Ferdous

Keyword(s):

The Netherlands ◽

Molecular Characterization ◽

Shiga Toxin ◽

E Coli

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Commensal Flora Reduce E. coli O157:H7 and Shiga Toxin in Mouse Intestine

Microbe Magazine ◽

10.1128/microbe.1.193.2 ◽

2006 ◽

Vol 1 (4) ◽

pp. 193-193

Keyword(s):

Shiga Toxin ◽

E Coli ◽

Commensal Flora ◽

Mouse Intestine

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Multiresistant Bacteria Isolated from Intestinal Faeces of Farm Animals in Austria

Antibiotics ◽

10.3390/antibiotics10040466 ◽

2021 ◽

Vol 10 (4) ◽

pp. 466

Author(s):

Herbert Galler ◽

Josefa Luxner ◽

Christian Petternel ◽

Franz F. Reinthaler ◽

Juliana Habib ◽

...

Keyword(s):

Meat Products ◽

Animal Husbandry ◽

Multidrug Resistant ◽

Farm Animals ◽

Resistant Bacteria ◽

Antibiotic Resistant Bacteria ◽

Antibiotic Resistant ◽

E Coli ◽

Multiresistant Bacteria ◽

Vancomycin Resistant

In recent years, antibiotic-resistant bacteria with an impact on human health, such as extended spectrum β-lactamase (ESBL)-containing Enterobacteriaceae, methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci (VRE), have become more common in food. This is due to the use of antibiotics in animal husbandry, which leads to the promotion of antibiotic resistance and thus also makes food a source of such resistant bacteria. Most studies dealing with this issue usually focus on the animals or processed food products to examine the antibiotic resistant bacteria. This study investigated the intestine as another main habitat besides the skin for multiresistant bacteria. For this purpose, faeces samples were taken directly from the intestines of swine (n = 71) and broiler (n = 100) during the slaughter process and analysed. All samples were from animals fed in Austria and slaughtered in Austrian slaughterhouses for food production. The samples were examined for the presence of ESBL-producing Enterobacteriaceae, MRSA, MRCoNS and VRE. The resistance genes of the isolated bacteria were detected and sequenced by PCR. Phenotypic ESBL-producing Escherichia coli could be isolated in 10% of broiler casings (10 out of 100) and 43.6% of swine casings (31 out of 71). In line with previous studies, the results of this study showed that CTX-M-1 was the dominant ESBL produced by E. coli from swine (n = 25, 83.3%) and SHV-12 from broilers (n = 13, 81.3%). Overall, the frequency of positive samples with multidrug-resistant bacteria was lower than in most comparable studies focusing on meat products.

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Utilizing the Food–Pathogen Metabolome to Putatively Identify Biomarkers for the Detection of Shiga Toxin-Producing E. coli (STEC) from Spinach

Metabolites ◽

10.3390/metabo11020067 ◽

2021 ◽

Vol 11 (2) ◽

pp. 67

Author(s):

Snehal R. Jadhav ◽

Rohan M. Shah ◽

Avinash V. Karpe ◽

Robert S. Barlow ◽

Kate E. McMillan ◽

...

Keyword(s):

Shiga Toxin ◽

Metabolic Profiling ◽

Biomarker Discovery ◽

Risk Groups ◽

Proof Of Concept ◽

Rapid Methods ◽

E Coli ◽

Leafy Greens ◽

Food Pathogen ◽

Genomic Similarity

Shiga toxigenic E. coli (STEC) are an important cause of foodborne disease globally with many outbreaks linked to the consumption of contaminated foods such as leafy greens. Existing methods for STEC detection and isolation are time-consuming. Rapid methods may assist in preventing contaminated products from reaching consumers. This proof-of-concept study aimed to determine if a metabolomics approach could be used to detect STEC contamination in spinach. Using untargeted metabolic profiling, the bacterial pellets and supernatants arising from bacterial and inoculated spinach enrichments were investigated for the presence of unique metabolites that enabled categorization of three E. coli risk groups. A total of 109 and 471 metabolite features were identified in bacterial and inoculated spinach enrichments, respectively. Supervised OPLS-DA analysis demonstrated clear discrimination between bacterial enrichments containing different risk groups. Further analysis of the spinach enrichments determined that pathogen risk groups 1 and 2 could be easily discriminated from the other groups, though some clustering of risk groups 1 and 2 was observed, likely representing their genomic similarity. Biomarker discovery identified metabolites that were significantly associated with risk groups and may be appropriate targets for potential biosensor development. This study has confirmed that metabolomics can be used to identify the presence of pathogenic E. coli likely to be implicated in human disease.

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W1780 Ciprofloxacin in Rabbits Infected with a Shiga-Toxin (Stx)-Producing E. coli (Stec) Srain Leads to Worse Clinical Outcome and Increased Toxin Release

Gastroenterology ◽

10.1016/s0016-5085(08)63334-x ◽

2008 ◽

Vol 134 (4) ◽

pp. A-714

Author(s):

Antonio Serna ◽

Chengru Zhu ◽

Amelia J. Nugent ◽

Erin K. Okeefe ◽

Edgar Boedeker

Keyword(s):

Clinical Outcome ◽

Shiga Toxin ◽

E Coli

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Hemoconcentration and predictors in Shiga toxin-producing E. coli-hemolytic uremic syndrome (STEC-HUS)

Pediatric Nephrology ◽

10.1007/s00467-021-05108-6 ◽

2021 ◽

Author(s):

Sebastian Loos ◽

Jun Oh ◽

Laura van de Loo ◽

Markus J. Kemper ◽

Martin Blohm ◽

...

Keyword(s):

Risk Factor ◽

Serum Creatinine ◽

Hemolytic Uremic Syndrome ◽

Shiga Toxin ◽

Neurological Symptoms ◽

Fluid Loss ◽

Good Prediction ◽

E Coli ◽

Complicated Course ◽

Uremic Syndrome

Abstract Background Hemoconcentration has been identified as a risk factor for a complicated course in Shiga toxin-producing E. coli-hemolytic uremic syndrome (STEC-HUS). This single-center study assesses hemoconcentration and predictors at presentation in STEC-HUS treated from 2009–2017. Methods Data of 107 pediatric patients with STEC-HUS were analyzed retrospectively. Patients with mild HUS (mHUS, definition: max. serum creatinine < 1.5 mg/dL and no major neurological symptoms) were compared to patients with severe HUS (sHUS, definition: max. serum creatinine ≥ 1.5 mg/dL ± major neurological symptoms). Additionally, predictors of complicated HUS (dialysis ± major neurological symptoms) were analyzed. Results Sixteen of one hundred seven (15%) patients had mHUS. Admission of patients with sHUS occurred median 2 days earlier after the onset of symptoms than in patients with mHUS. On admission, patients with subsequent sHUS had significantly higher median hemoglobin (9.5 g/dL (3.6–15.7) vs. 8.5 g/dL (4.2–11.5), p = 0.016) than patients with mHUS. The product of hemoglobin (g/dL) and LDH (U/L) (cutoff value 13,302, sensitivity 78.0%, specificity of 87.5%) was a predictor of severe vs. mild HUS. Creatinine (AUC 0.86, 95% CI 0.79–0.93) and the previously published score hemoglobin (g/dL) + 2 × creatinine (mg/dL) showed a good prediction for development of complicated HUS (AUC 0.87, 95% CI 0.80–0.93). Conclusions At presentation, patients with subsequent severe STEC-HUS had a higher degree of hemoconcentration. This underlines that fluid loss or reduced fluid intake/administration may be a risk factor for severe HUS. The good predictive value of the score hemoglobin (g/dL) + 2 × creatinine (mg/dL) for complicated HUS could be validated in our cohort. Graphical abstract

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