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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.

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 Pearsons 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. [J Adv Vet Anim Res 2018; 5(2.000): 218-225]

Milkborne transmission of Shiga toxin- producing Escherichia coli (STEC) has raised considerable concern due to recent outbreaks worldwide and poses a threat to public health. The aim of this study was to develop a sensitive and specific multiplex PCR assay to detect the presence of STEC in bovine raw milk. To identify E. coli (ATCC 25922) contamination, the gene uspA was used, and PCR sensitivity and specificity were accessed by testing diluted samples ranging from 2 to 2.0 × 10(6) CFU/mL. To detect STEC, the stx1 and stx2 genes were selected as targets. After reaction standardization, the multiplex assay was tested in raw milk collected from 101 cows on dairy farms. PCR assay for E. coli detection had a specificity of 100% and sensitivity of 79% (P

Shiga toxin-producing Escherichia coli (STEC) and enterotoxigenic E. coli (ETEC) are important causes of diarrhea in humans and animals worldwide. Although ruminant animals are the main source of STEC, diarrhea due to this pathotype is very low in Bangladesh where ETEC remains the predominant group associated with childhood diarrhea. In the present study, E. coli strains (n = 35) isolated from Bangladesh livestock (goats, sheep, and cattle) and poultry (chicken and ducks) were analyzed for the presence of major virulence factors, such as Shiga toxins (STX-1 and STX-2), heat-labile toxin, and heat-stable toxins (STa and STb). Multiplex polymerase chain reaction results revealed 23 (66%) E. coli strains to be virulent possessing either sta (n = 5), stx (stx1, n = 8; stx2, n = 2), or both (n = 8) genes in varying combinations. Thirty-four percent (8/23) of strains from livestock were hybrid type that carried both stx (either stx1 or stx2) and ETEC-specific enterotoxin gene sta. Serotyping results revealed that the ETEC strains belonged to five serotypes, namely O36: H5, O174: H-, O152: H8, O109: H51, and O8: H21, while the STEC-producing strains belonged to serotypes O76: H19 (n = 3), O43: H2 (n = 2), O87: H16 (n = 2), OR: H2 (n = 1), O110: H16 (n = 1), and O152: H8 (n = 1). The STEC-ETEC hybrid strains belonged to serotypes O76: H19 (n = 3), O43: H2 (n = 2), O87: H16, OR: H2, and O152: H8. Forty percent (2/5) of the ETEC and 20% (2/10) of the STEC strains were multidrug resistant with the highest drug resistance (50%) being found in the hybrid strains. Molecular fingerprinting determined by pulsed-field gel electrophoresis and cluster analyses by dendrogram revealed that, genetically, STEC-ETEC hybrid strains were highly heterogeneous. Multidrug-resistant E. coli STEC-ETEC hybrid strains in domesticated animals pose a public health threat for humans in Bangladesh.

Shiga toxins (Stx1 and Stx2) are the main virulence factors of enterohemorrhagic Escherichia coli (EHEC), a foodborne pathogen associated with diarrhea, hemorrhagic colitis and hemolytic uremic syndrome. The aim of this study was to evaluate the antibodies against Stx2 obtained from egg yolks of laying hens immunized with a recombinant Stx2B subunit. A high specific response in serum was observed 25 days after the first immunization and IgY antibodies were extracted from day 47th and purified from egg yolk. A concentration of 0.84 mg of total IgY/ml of egg yolk was obtained, of which 8% were antigen specific. The ability of anti-Stx2B IgY to recognize Stx2B and Stx2 either in solid-phase or in solution were evaluated and compared with anti-Stx2B rabbit antibodies by Western blotting and ELISA. The protective efficacy of IgY against Stx2 was determined by in vitro and in vivo experiments. The results show that IgY was able to recognize Stx2B and Stx2 in denatured conditions, attached to a solid-phase and free in solution. The anti-Stx2B IgY could effectively block the biological activity of Stx2 on Vero cells and protect mice from Stx2 challenge. The data suggest that immunization of hens with Stx2B could be a strategy to obtain at low cost a relatively high concentration of anti-Stx2 egg yolk IgY, able to neutralize Stx2 lethal activity. IgY technology could be an useful tool for research, diagnosis and therapy of EHEC infection., Highlights ► We studied the antibodies response in laying hens against Shiga toxin 2 B subunit. ► IgY antibodies anti-Shiga toxin 2 are obtained from egg yolk. ► Specific IgY binds to Shiga toxin 2 and neutralize activity in vivo and in vitro.

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. Fil: Alonso, Mónica Zulema. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Lucchesi, Paula Maria Alejandra. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Rodriguez, Edgardo Mario. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Parma, Alberto Ernesto. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Padola, Nora Lía. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-27T11:25:57Z No. of bitstreams: 0Bitstream added on 2014-05-27T14:36:19Z : No. of bitstreams: 1 2-s2.0-79961068787.pdf: 846743 bytes, checksum: 44ff5a4c4c6011063fd4120ed61ea6e2 (MD5) Made available in DSpace on 2014-05-27T11:25:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-08-08 Bacterial cultures of cloaca swabs from 86 captivity kept psittacidaes revealed 17 Escherichia coli bearing birds sharing strains which, on the basis of enterobacterial repetitive intergenic consensus (ERIC) PCR analysis, proved to be genetically similar. Further, triplex PCR specific for the genetic markers chuA, yjaA, and TSPE4.C2 was used to assign the strains to the E. coli reference collection (EcoR) B2 group. One strain of each, from the enteropathogenic (EPEC), enteroaggregative (EAEC) and Shiga toxin (STEC) E. coli pathovars were found among these isolates. © Marietto-Gonçalves et al.; Licensee Bentham Open. Department of Microbiology and Immunology Institute of Bioscience of the São Paulo State University in Botucatu, ZIP 18618-970, Botucatu SP Floravida Institute, Botucatu SP Department of Microbiology and Immunology Institute of Bioscience of the São Paulo State University in Botucatu, ZIP 18618-970, Botucatu SP

Aims: To demonstrate that a thorough characterization and virulotyping of Escherichia coli strains isolated from sheep over time leads to new insights into ovine E. coli potentially becoming human pathogens through horizontal gene transfer. Methods and Results: One hundred and fifty E. coli isolates from two sheep, sampled over 3 weeks, were characterized by serotyping, virulotyping, genotyping using multiple locus variable number tandem repeats analysis (MLVA) and susceptibility to phage infection in vitro. The 35 MLVA profiles and the serotype and virulotypes of the strains were closely associated. Many MLVA profiles differed in one locus independent of serotypes. Escherichia coli isolates of the same serotype or virulotype had identical or very similar MLVA profiles. No transductants that incorporated the bacteriophages were found in vivo, but six E. coli isolates were susceptible to the phage infection in vitro. Changes in MLVA profiles were seen after acquisition of Stx phages in vitro only. Conclusions: The sheep carried Stx phage susceptible E. coli that possessed virulence markers associated with human pathogenicity. Changes in bacterial genomes by phage transfer may complicate outbreak source investigations. Serotype has to be taken into account when evaluating strain relationships by MLVA. Significance and Impact of the Study: Sheep carry E. coli that encode for virulence markers and belong to serogroups known to be human pathogens. In addition, a selection of isolates was found to be susceptible to horizontal transfer of Shiga toxin genes by means of bacteriophages in vitro, and the transfer resulted in a discernible change of the MLVA patterns of E. coli.