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High hydrostatic pressure processing (HHP) is used to reduce or eliminate pathogens and spoilage bacteria in food. The aim of this study was to investigate the effect of fat on the pressure resistance of E. coli in beef and a yogurt model system. Ground beef and pH-adjusted (pH 5.5) yogurt were adjusted to a fat content from 3 to 35% with adipose tissue and crème fraiche, respectively, and inoculated with E. coli. Samples were treated at 600 MPa and 20 or 30 °C for 3 min. Sample temperatures were adjusted prior to compression to compensate adiabatic heating, or not. An increased fat content of ground beef decreased the pressure resistance of E. coli. This was observed irrespective of the temperature history of the treatment. For example, at 20 °C, an increase in fat content from 15.5 to 24.4% increased treatment lethality by 6 log CFU/g for E. coli MG 1655. The correction for adiabatic heating increased or decreased pressure resistance at 20 °C in a strain-specific manner. Addition of crème fraiche to pH-adjusted yogurt did not alter the pressure resistance of E. coli (strains MG 1655 and DM 18.3), or increased resistance (E. coli AW 1.7 and AW 1.3). In summary, the pressure resistance of E. coli is dependent on the fat content, the food matrix and the temperature trajectory during processing. The effect of adipose tissue in meat matrix links the fat content of meat to lipid oxidation and the oxidative stress response. [ABSTRACT FROM AUTHOR]

Generally, the majority of the food poisoning crisis from seafood comes out due to shellfish consumption, mainly gandofflibecause ofits filter feeders pattern. Many microbes are used as an indicator of the hygienic status of several foods, one of them is Escherichia coli which is used to detect fecal pollution in water and shellfish. Therefore, twenty gandoffli samples were randomly collected from local markets in Ismailia city, Egypt, for evaluation of Enterobacteriaceae counts, and identification of Escherichia coli and detection of phoA and Shiga toxin genes. The obtained results revealed that the total Enterobacteriaceae count of the gandoffli ranged from 7x10² to 7x105 cfu/g with an average of 5x104±3.5x104cfu/g. The occurrence of Enterobacteriaceae members in gandoffli was represented by E. coli (99%), Acinetobacter lwoffii (99%), Enterobacter hormaechei (85%), Klebsiella oxytoca (95%), Stentrophomonas maltophilia (85%), Moraxella lacunata (85%), Achromobacter xylosoxidans (93%), and ESBL E. coli (100%). In addition, E. coli isolated from gandoffli were subsequently serologically typed into O103, O55, O 128, O 126, and O157 then confirmed using conventional polymerase chain reaction by the presence of alkaline phosphatase gene. Upon checking virulence genes in E. coli: stx2 was absent in O157 and O103. Also, stx1 was present in O157 and absent in O103. Its should be concluded that gandoffli were exposed to Enterobacteriaceae contamination from different sources during handling, storage and distribution. Gandoffli had E. coli and their toxin that can pose serious public health hazards to consumers. Strick hygienic measures must be applied through the chain of gandoffli production to ensure their safety for consumer consumptions [ABSTRACT FROM AUTHOR]

Despite the lack of scientific evidence, some consumers assert that raw milk is a natural food with nutritional and immunological properties superior to pasteurized milk. This has led to the increased popularity of unpasteurized cow milk (UPM) and disregard for the risks of being exposed to zoonotic infections. Dairy cattle are healthy carriers of Shiga toxin (Stx)-producing E. coli (STEC), and contaminated UPM has caused STEC outbreaks worldwide. The association between STEC, carrying the eae (E. coli attachment effacement) gene, and severe diseases is well-established. We have previously isolated four eae positive STEC isolates from two neighboring dairy farms in the Southeast of Norway. A whole genome analysis revealed that isolates from different farms exhibited nearly identical genetic profiles. To explore the risks associated with drinking UPM, we examined the ability of the isolates to produce Stx and their growth in UPM at different temperatures. All the isolates produced Stx and one of the isolates was able to propagate in UPM at 8 °C (p < 0.02). Altogether, these results highlight the risk for STEC infections associated with the consumption of UPM. [ABSTRACT FROM AUTHOR]

Oedema disease (ED) caused by Shiga-toxin-producing E. coli in pigs is a serious life-threatening disease, particularly among weaned piglets. When a preventive protocol is adopted in a specific farm, interpretation of effectiveness is often complicated in field conditions due to natural or “common cause” variation. For this reason, in this study a Statistical process control (SPC) approach was used to retrospectively evaluate the application of an ED preventive protocol (lower protein diet, ad-libitum fiber, vaccination at 5 days of age) in an infected commercial piglets' weaning site. The analysis was established over a 9-years period (n = 75 consecutive batches; 1,800 weaners per batch) using mortality for each batch as the key parameter of health and production; the statistics and the control limits (mean ± 3-fold sd; UCL, upper control limit; LCL, lower control limit) were based on data from the first 28 batches (Period 1) before the onset of the first ED clinical signs. The charts allowed the detection of defined out of control batches (i.e., with mortality out of the intervention limits) from batch 29 ongoing, exploring a Period 2 (unstable production and ED clinical signs; 36 batches) and a Period 3 (application of the ED preventive protocol; 11 batches). Mortality evaluation using SPC revealed a production system defined under-control (mean moving range bar = 1,34%; UCL = 4,37%; LCL = 0%) during Period 1. During Period 2, charts lost the state of statistical control, as showed by several signals of special cause variation due to the ED outbreak. Period 3 was characterized again by a state of statistical control, where no signals of special cause variation was showed. In conclusion, the retrospective application of SPC charts in the present study was able to confirm the efficacy of an ED preventive protocol in reducing mortality in a piglets' weaning site. SPC charting is suggested as an useful tool to provide insights into relationships between health, managerial, and welfare decision and some selected iceberg parameters in livestock.

Background and Objectives: Enterohemorrhagic Escherichia coli (EHEC) causes bloody and non-bloody diarrhea, intestinal infection and extraintestinal complications in humans. This study aimed to isolate and evaluate the prevalence of E. coli O157: H7 and other Shiga toxin-producing E. coli (STEC) and identify the virulence genes (stx1, stx2, hly and eaeA) from patients with diarrhea. Also, the antibiotic resistance profile of the isolated strains was evaluated. Materials and Methods: A total of 100 stool samples were collected from patients with acute diarrhea referring to the hospital and clinics in Isfahan County, Iran. Phenotypic tests and PCR assay were used for detection of E. coli O157: H7 and other Shiga toxin-producing E. coli. The presence of virulence genes (stx1, stx2, hly and eaeA) were identified by PCR. The antibiotic resistance profile of the isolates was determined using the agar disk diffusion method. The results were analyzed descriptively by Sigma stat version 4 software. Results: Seventy - eight out of 100 samples (78%) were contaminated with E. coli. E. coli O157 was isolated from five samples (6.4%), of which only two strains (2.56%) were identified as E. coli O157: H7. According to the results, out of two E. coli O157: H7 isolates, one (50%) isolate contained eaeA and two isolates (100%) contained Stx1, Stx2, hlyA genes. Out of three (3.84%) E. coli O157: HN, one of the isolate (33.3%) contained stx1 and, two isolates (66.7%) were positive for hlyA genes. Also, the results revealed that six strains (7.69%) were non-O157: H7 STEC, of which two isolates (33.3%) contained stx1 and four isolates (66.7%) were positive for stx2 and hlyA genes. The results of antibiogram tests revealed that all of the STEC isolates (100%) were sensitive to imipenem followed by kanamycin, gentamicin and nitrofurantoin (91%). High resistance (54.5%) to ampicillin and ciprofloxacin was observed among the STEC isolates. Conclusion: The results of the current study showed that although the prevalence of E. coli O157: H7 was low among patients with diarrhea, the other STEC strains with relative resistance to antibiotics are more prevalent.

Background and Objectives: Enterohemorrhagic Escherichia coli (EHEC) causes bloody and non-bloody diarrhea, intestinal infection and extraintestinal complications in humans. This study aimed to isolate and evaluate the prevalence of E. coli O157: H7 and other Shiga toxin-producing E. coli (STEC) and identify the virulence genes (stx1, stx2, hly and eaeA) from patients with diarrhea. Also, the antibiotic resistance profile of the isolated strains was evaluated. Materials and Methods: A total of 100 stool samples were collected from patients with acute diarrhea referring to the hospital and clinics in Isfahan County, Iran. Phenotypic tests and PCR assay were used for detection of E. coli O157: H7 and other Shiga toxin-producing E. coli. The presence of virulence genes (stx1, stx2, hly and eaeA) were identified by PCR. The antibiotic resistance profile of the isolates was determined using the agar disk diffusion method. The results were analyzed descriptively by Sigma stat version 4 software. Results: Seventy - eight out of 100 samples (78%) were contaminated with E. coli. E. coli O157 was isolated from five samples (6.4%), of which only two strains (2.56%) were identified as E. coli O157: H7. According to the results, out of two E. coli O157: H7 isolates, one (50%) isolate contained eaeA and two isolates (100%) contained Stx1, Stx2, hlyA genes. Out of three (3.84%) E. coli O157: HN, one of the isolate (33.3%) contained stx1 and, two isolates (66.7%) were positive for hlyA genes. Also, the results revealed that six strains (7.69%) were non-O157: H7 STEC, of which two isolates (33.3%) contained stx1 and four isolates (66.7%) were positive for stx2 and hlyA genes. The results of antibiogram tests revealed that all of the STEC isolates (100%) were sensitive to imipenem followed by kanamycin, gentamicin and nitrofurantoin (91%). High resistance (54.5%) to ampicillin and ciprofloxacin was observed among the STEC isolates. Conclusion: The results of the current study showed that although the prevalence of E. coli O157: H7 was low among patients with diarrhea, the other STEC strains with relative resistance to antibiotics are more prevalent. [ABSTRACT FROM AUTHOR]

Objective: Shiga-toxin producing Escherichia coli (STEC) is the most important foodborne bacterial pathogen worldwide and the bovine animals are assumed as a reservoir of this pathogen. The present study was conducted to assess the role of bovine animals as the source of STEC. Materials and methods: To assess the role of bovine animals as the source of STEC, we examined 100 samples (50 rectal swab and 50 beef samples) collected from the local beef slaughterhouses by cultural, morphological, biochemical tests and polymerase chain reaction. Finally, the drug resistance pattern of isolated organisms has been examined. Result: In the preliminary screening by polymerase chain reaction (PCR), E. coli was more prevalent in rectal swab (n=21/50) than beef samples (n=16/50). Among 39 isolated E. coli, 10 isolates were confirmed as STEC (Rectal swab=7, Beef=3) by PCR, where stx2 gene (n=7/10) was predominant than stx1 gene (n=3/10). Remaining 29 isolates did not react to stx primers in PCR. Presence of STEC in beef samples was significantly associated with the fecal contamination at P≤0.1 (0.074818) in Pearson's correlation coefficient method. In addition, most of the isolated STEC strains were resistant to one or more commonly used antimicrobials in the country. Conclusion: The bovine animals and its products could be an important source of multidrug-resistant STEC in the country. [ABSTRACT FROM AUTHOR]

An investigation of mortality of piglets through clinical signs, post-mortem, histopathology and bacteriological analyses revealed the causal organism to be Escherichia coli, mainly O149:K91:K88 which belongs to the enterotoxigenic biotypes. Molecular characterization and epidemiologic analysis elucidated it as shiga-toxin (ST) E. coli resistant to ampicillin, cefotaxime, tetracycline, trimethoprim-sulfamethoxazole, tylosin and neomycin. Conventional PCR results detected genes for ST-2, adhesin involved in diffuse adherence (AIDA-1) and F18 fimbriae virulence factors. Survival analyses and logistic regression of piglet mortality patterns showed that season of weaning, weaning weight and age of dam had significant influence on survival rate of piglets. Factors affecting pathogenicity of bowel edema and survival of affected piglets on a farm with persistent infection were reported for the first time. An association of E. coli O149:K91:K88 (F4) with clinical edema disease was made even though it has been reported in the past that this serotype does not produce ST. It was concluded that more stringent measures to mitigate the impact of the disease need to be targeted for spring and in older sows. [ABSTRACT FROM AUTHOR]

Abstract: To better understand Escherichia coli O157:H7 on-farm transmission dynamics requires sensitive methods for quantification of a broad range of concentrations of target organisms. For this purpose, a multiplex real time PCR (qPCR) assay was developed for quantification of O157 E. coli from 1g fecal samples of cattle and other animal species, targeting the Shiga toxin genes (stx1 and stx2) and the O157 somatic antigen gene, per. The multiplex qPCR assay provided specific detection across a broad range of bacterial concentrations with a lower limit of detection (LOD) of 101 genome copies which is equivalent to 101 bacteria. However, the LOD, when direct qPCR was applied to quantification of the targets in the feces of dairy cattle, was 103 genome copies per gram of feces. Enumeration below the threshold for direct qPCR was performed using a modified most probable number (mMPN) method whereby E. coli O157 in enriched samples was isolated using immunomagnetic bead separation (IMS) and detected using qPCR, thus reducing the time and logistic constraints of biochemical/serological/gel analysis. Application of the mMPN (IMS/qPCR) assay to samples that were negative when tested using direct qPCR alone permitted quantification of low levels of E. coli O157 below levels detectable with direct qPCR. The direct qPCR and mMPN (IMS/qPCR) assays were applied to fecal samples from dairy, beef, swine and poultry feces. This approach can be employed to gain a better understanding of the patterns of infection in animals for analysis of on-farm transmission dynamics, for evaluating the effects of on-farm control strategies and for risk assessment in public health. [Copyright &y& Elsevier]

Foodborne diseases are mainly caused by bacterial contamination which can lead to severe diarrhea. This study aimed to detect the presence of Shiga toxin-Producing Escherichia coli O157, Escherichia coli non-O157 and virulence gene in raw vegetables. The samples were purchased from wet market and hypermarket in Selangor. The detections were carried out by using the combination methods of Most Probable Number-Polymerase Chain Reaction (MPNPCR). A total of 37(18.5%) samples were found to be contaminated by STEC. Out of these 37 isolates, four (10.8%) of the isolates were E. coli O157 while 33(89.2%) were E. coli non-O157. However, there was no E. coli O157:H7 detected in all the samples. The occurrence of Shiga toxin-Producing E. coli in edible raw vegetables samples suggests the importance of this pathogen in vegetables. Therefore, more studies are required to remove this pathogen from vegetables. [ABSTRACT FROM AUTHOR]

Copyright of Reanimation is the property of Lavoisier and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Abstract: Shiga-toxigenic strains of Escherichia coli (STEC) are zoonotic pathogens with human health, meat processing and trade impacts. Cattle are the principal reservoirs of STEC, although other animals can be carriers. The STEC status of Australian native marsupials has not been formatively described to date. The aim of the current study was to investigate carriage of STEC by native Australian marsupials in Southeast Queensland. Faeces from a variety of marsupial species, stratified by gastrointestinal morphology and dietary type, were screened for stx 1, stx 2 and other STEC virulence genes by PCR. Positive samples were cultured to isolate STEC for characterisation. A number of macropods from both captive and wild habitats had evidence of STEC in their faeces. Rates of stx carriage by macropods (8.6%) were comparable, though generally low, compared to cattle. Eastern grey kangaroos had the highest rate of stx presence in faeces (10.3%). Hindgut-fermenting and monogastric marsupials had no evidence of STEC shedding. Based on virulence marker possession and serotype, the human pathogenic potential of isolates was low. This is the first report of Australian marsupials carrying STEC. Australian native macropods may act as reservoirs for STEC strains, but the potential significance to public health and/or livestock epidemiology remains questionable. [Copyright &y& Elsevier]

Abstract: Enteropathogenic (EPEC) and Shigatoxigenic Escherichia coli (STEC) are foodborne pathogens that cause potentially fatal infant diarrhea and hemolytic uremic syndrome, respectively. We investigated the presence of intimin and Shiga toxin encoding genes, as indicators of EPEC and STEC presence in cloacae and chicken products. The analyzed products were hamburgers, giblets and carcasses obtained from poultry and butcher shops. EPEC contamination predominated over STEC contamination in cloacae and chicken products, although some differences were detected when the kind of food or shop was taken into account. In particular, among chicken hamburgers we found a greater proportion of EPEC than STEC-positive samples at poultry shops, while in butcheries STEC was predominant. This finding could suggest cross contamination during handling at butcheries. The results indicate that it is necessary to improve hygienic measures both during slaughtering and manipulation of chicken products at retail stores, to provide a safe product to consumers. [Copyright &y& Elsevier]

Abstract: Probiotics are known to have an inhibitory effect against the growth of various foodborne pathogens, however, the specific role of probiotics in Shiga-toxin-producing Escherichia coli (STEC) virulence gene expression has not been well defined. Shiga toxins are members of a family of highly potent bacterial toxins and are the main virulence marker for STEC. Shiga toxins inhibit protein synthesis in eukaryotic cells and play a role in hemorrhagic colitis and hemolytic uremic syndrome. STEC possesses Shiga toxin 1 (Stx1) and Shiga toxin 2 (Stx2), both of which have A and B subunits. Although STEC containing both Stx1 and Stx2 has been isolated from patients with hemorrhagic colitis, Stx2 is more frequently associated with human disease complications. Thus, the effect of Lactobacillus, Pediococcus, and Bifidobacterium strains on stx2A expression levels in STEC was investigated. Lactic acid bacteria and bifidobacteria were isolated from farm animals, dairy, and human sources and included L. rhamnosus GG, L. curvatus, L. plantarum, L. jensenii, L. acidophilus, L. casei, L. reuteri, P. acidilactici, P. cerevisiae, P. pentosaceus, B. thermophilum, B. boum, B. suis and B. animalis. E. coli O157:H7 (EDL 933) was coincubated with sub-lethal concentrations of each probiotic strain. Following RNA extraction and cDNA synthesis, relative stx2A mRNA levels were determined according to a comparative critical threshold (Ct) real-time PCR. Data were normalized to the endogenous control glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and the level of stx2A expression between treated and untreated STEC was compared. Observed for all probiotic strains tested, stx2A was down-regulated, when compared to the control culture. Probiotic production of organic acids, as demonstrated by a decrease in pH, influenced stx2A gene expression. [Copyright &y& Elsevier]

Aim: To quantify the effect of enrichment, immunomagnetic separation (IMS), and selective plating procedures on isolation of Shiga-toxigenic Escherichia coli O157 (STEC O157) and non-Shiga-toxigenic Escherichia coli O157 (non-STEC O157) from naturally contaminated bovine faeces. Methods and Results: Two broth enrichment times, two IMS strategies, and two selective plating media were evaluated. STEC O157 and non-STEC O157 strains were often isolated from the same faecal specimen and responded differently to the isolation protocols. A large-volume IMS system was more sensitive than a conventional small-volume IMS method, but was also more expensive. STEC O157 was more frequently isolated from 6 h enriched broth and ChromAgar plates containing 0·63 mg l−1 potassium tellurite (TCA). Non-STEC O157 was more frequently isolated from un-enriched broth and ChromAgar plates without tellurite (CA). Conclusions: The combination of 6-h enrichment in Gram-negative broth containing vancomycin, cefixime and cefsuludin, large volume IMS and selective plating on TCA maximized STEC O157 recovery from naturally contaminated cattle faecal specimens. Significance and Impact of the Study: The pairing of proper enrichment with a specific plating procedure is key for STEC O157 recovery from naturally contaminated bovine faeces. Incorporating tellurite into an E. coli O157 detection strategy may select for the subset of E. coli O157 that contains the Shiga-toxin genes. [ABSTRACT FROM AUTHOR]

Copyright of Revista Chilena de Pediatría is the property of Revista Chilena de Pediatria and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Aims: To review and characterize the enrichment protocols used for detecting all Shiga-Toxin producing Escherichia coli (STEC) from different matrices. Methods and Results: Firstly, the frequency distribution of the factors characterizing the enrichment protocols is described; secondly, a multiple correspondence analysis is performed to display profiles of association of these factors, and thirdly, published results concerning the relative performances of the protocols are summarized. Trypticase Soy Broth (TSB) is reported as the most frequently used enrichment broth. More often, one antibiotic is added in enrichment broths and these broths are incubated for a duration of 16–24 h at 35–37°C. It also appears that the incubation temperature does not seem to be related to the type of serogroup looked for and that antibiotics are used regardless of the matrix analysed. Finally, results relating to the enrichment protocol efficacy are rare and differ from one study to another. Conclusions: Statistical studies must be conducted so as to assess the efficacy of the main enrichment protocols investigated in this study. Significance and Impact of the Study: This study reviews the most commonly used enrichment protocols and highlights the lack of results as to their relative efficacy. [ABSTRACT FROM AUTHOR]

Only a few studies concerning Shiga toxin-producing E. coli (STEC) detection in bivalves and their harvesting areas have been reported, and to the best of our knowledge there are no outbreaks associated with STEC from bivalves described. The aim of the present study was to investigate the occurrence of STEC in Norwegian bivalves, and to characterize potential STEC isolated from the samples. A total of 269 samples of bivalves were screened for the presence of stx and eae genes, and markers for the serogroups O26, O103, O111, O145 and O157 by using ISO TS 13136 (2012). The screening returned 19 samples that were positive for stx and eae , and attempts of isolation of STEC were made from these samples. Presumptive STEC were obtained from three samples, and three isolates (one from each sample) were subjected to whole-genome-sequencing (WGS). The WGS revealed that one of the isolates did not carry the stx genes, while the other two were identified as stx 2i positive E. coli O9:H19 and stx 2g positive E. coli O96:H19. Neither of the two STEC isolates were positive for virulence markers such as eae and ehx. The results suggest that the occurrence of STEC in Norwegian bivalves is low. • First time in Norway that STEC has been detected in bivalves. • Bivalves could be a potential vehicle for transmission of STEC. • One strain harboured a newly described Shiga toxin variant; stx 2 i. • Hybrid STEC O96:H19 isolated from shellfish (stx 2g and ST-Ia) with newly identified MLST type. [ABSTRACT FROM AUTHOR]

Background: Haemolytic uraemic syndrome is a rarely seen in adults often leading to critical illness. This case highlights how difficult it can be to establish a diagnosis and treat when a patient presents with bloody diarrhoea.Case Presentation: A 17-year-old Iraqi man presented to the emergency department with abdominal pain and bloody diarrhoea. He was initially treated as acute appendicitis, undergoing an appendectomy but following a recurrence in his symptoms a colonoscopy was performed. A diagnosis of shiga toxin-producing Escherichia coli leading to HUS was suspected following histology obtained at colonoscopy and this was confirmed on antibody testing. Despite intravenous fluids and supportive therapy the patient's symptoms and condition deteriorated. He developed seizures and acute renal failure requiring intubation and plasma exchange in the intensive care setting. He eventually required treatment with ecluzimab therapy; a monoclonal antibody and subsequently made a full recovery.Conclusions: Haemolytic uraemic syndrome is a triad of progressive renal failure, thrombocytopenia and haemolytic anaemia which is a condition rarely seen in adults. It is usually associated with an E. coli infection and supportive therapy remains the mainstay of treatment. [ABSTRACT FROM AUTHOR]