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This book presents an overview of Shiga toxin-producing E. coli (STEC), with in-depth coverage of key areas such as recent Shiga toxin-related poisonings in Europe and the US, the structure, production, and mechanism of action of Shiga toxin, and current methods of detection. The globalization of food production has introduced new risk factors and intensified existing hazards, complicating the assurance of food safety. Foodborne illness outbreaks, such as those related to STEC, are becoming more common and more dangerous. The threat that these bacterial toxins pose to the food supply is magnified by the frequent occurrence and severity of Shiga toxin-caused disease. As a result, STEC and their toxins remain a primary concern in food safety. This review serves as a key resource for scientists in the field and public health and regulatory officials charged with maintaining food safety. This book also looks to the future of treatment of Shiga toxin-associated disease, specifically the translation of lab bench science into clinical therapeutic strategies.
Shiga toxin-producing Escherichia coli (STEC) is an important foodborne pathogen associated with both outbreaks and sporadic cases of human disease, ranging from uncomplicated diarrhoea to haemorrhagic colitis (HC) and haemolytic uraemic syndrome (HUS). STEC affects children, elderly and immuno-compromised patients. STEC is capable of producing Shiga toxin type 1 (Stx1), type 2 (Stx2) or both, encoded by stx1 and stx2 genes, respectively. These strains are likely to produce putative accessory virulence factors such as intimin (encoded by eae), an enterohaemolysin (EhxA) and an autoagglutinating protein commonly associated with eae-negative strains (Saa), both encoded by an enterohaemorrhagic plasmid. Several studies have confirmed that cattle are the principal reservoir of STEC (O157 and non-O157:H7 serotypes) and many of these serotypes have been involved in HUS and HC outbreaks in other countries. Transmission of STEC to humans occurs through the consumption of undercooked meat, vegetables and water contaminated by faeces of carriers and by person-to-person contact. Diagnostic methods have evolved to avoid selective diagnostics, currently using molecular techniques for typing and subtyping of strains. Control is still a challenge, although there are animal vaccines directed against the serotype O157:H7.
Enterohemorrhagic E. coli (EHEC) are responsible for important diseases such as hemorrhagic colitis,hemolytic and uremic syndrome or thrombotic thrombocytopenic purpura. Human infections occurprincipally by consumption of contaminated food particularly beef meat, milk or vegetables. The mainnaturally reservoir of EHEC is the gastro-intestinal tract of cattle. Cattle feces are therefore responsiblefor contamination of various types of food but also environment dissemination of the pathogenicbacteria. Related to this problematic, the two objectives of this project were (i) to develop a newimmuno-capture method to improve the isolation of the main serogroups of EHEC involved in humaninfections in food; and (ii) to develop a new direct fed microbial usable in cattle to decrease prevalenceof EHEC in animals. This thesis allowed the development of an immuno-capture method based on theuse of 96-well microplates coated with specific antibody directed against E. coli O157 and otherserogroups. This method, called immuno-microplate capture (IMC) was efficient and user-friendly forthe isolation of E. coli O157; O26; O103 and O111 in foods. This could be an alternative to the use ofimmuno-magnetic beads which are currently used for the detection of EHEC in foods, but are timeconsumingand labor intensive when large number of samples is analyzed simultaneously. The secondpart of this thesis allowed the selection of 5 lactic acid bacteria strains which presented highantagonistic activity against E. coli O157 and other serogroups in vitro. Resistance of these strains togastro-intestinal conditions (acidic conditions, presence of bile salts and rumen fluid) was evaluated invitro. The safety of the 5 strains was checked in Balb-C mice by administration of each strain mixed infeed at 109 cfu/g. Finally, freeze-drying did not affect the antagonistic activity of the 5 strains, suggesteda possible large scale use of these strains. According to the various results obtained in vitro, the 5 strainscould potentially be used as DFM in cattle to decrease colonization of their gastro-intestinal tract byEHEC and consequently decrease the risk of food and environment contaminations.
Certain pathogenic Escherichia coli known as Shiga toxin (Stx)-producing Escherichia coli (STEC) are commensals in cattle, and typically cause bloody diarrhea in humans once the Stx toxin is secreted in invaded intestinal epithelial cells. Infections with STEC cells can lead to hemolytic uremic syndrome, which is commonly associated with kidney failure. Several STEC serogroups have been declared adulterants in raw, non-intact ground meat, and future regulations could potentially lead to a higher number of STEC serogroup detection strategies for these pathogenic microorganisms. Microbiological research laboratories may benefit from formalin-fixed STEC cells for periodic (daily, weekly, monthly, among others) instrument validation/calibration by serving as a working set of known cell concentration samples and internal standard i.e. positive control. These cell concentrations may be used across laboratories in different geographical locations, within an individual laboratory, and across a broad range of detection assays (molecular as well as immuno-based). This thesis consists of three research parts: a comprehensive literature review that covers STEC incidence in foods and molecular detection techniques (chapter 1), a literature review that covers immuno-based detection strategies (chapter 2), and a research manuscript that involves the development of an internal standard and positive control with formalin-fixed STEC cells that can be used for a broad range of molecular as well immuno-based detection assays for instrument calibration and validation purposes (chapter 3)
Escherichia coli O157:H7 and over 380 non-O157 serotypes of Shiga toxin producing E. coli (STEC) are human food-borne pathogens that inhabit the hindgut of ruminants and are shed in the feces, which subsequently contaminate food products. Recent epidemiological data have shown that six non-O157 STEC (O26, O103, O111, O121, O45 and O145) account for majority of human STEC infections. Fecal shedding of STEC is influenced by a number of factors, including diets, supplements, and feed additives, because of their potential to alter hindgut ecosystem. Not much is known about the fecal shedding of non-O157 STEC in cattle because of lack of standardized detection methods. Fecal shedding of E. coli O157:H7 was studied to determine the effects of supplemental urea, monensin, an ionophore, and ractopamine, a beta-agonist. Cattle fed monensin at 44 mg/kg of feed had lower (P = 0.05) fecal O157:H7 prevalence than cattle fed 33 mg/kg. Supplemental urea (0.35 or 0.70% of the diet) and inclusion of ractopamine at 200 mg/animal/day had no effect on fecal shedding of E. coli O157:H7. In an experimental inoculation study, inclusion of corn starch to a distiller's grains (DG)-supplemented diet had no effect on fecal shedding of E. coli O157 suggesting that either the decreased starch content in the DG-supplemented diet is not a factor in the increased shedding of E. coli O157:H7 or inclusion of pure starch in the diet may not have achieved our intended goal to have starch flow into the hindgut similar to that of corn grain. A multiplex PCR to detect O26, O45, O103, O111, O121, O145, and O157 was designed and applicability to detect the seven serogroups in cattle feces was evaluated. A multiplex PCR, designed to detect E. coli O104, feces showed presence of O104 in cattle feces (20.6%), but the isolated strains did not carry genes characteristic of the virulent strain responsible for the 2011 food-borne outbreak in Germany. Two preharvest interventions, a siderophore receptor and porin proteins-based vaccine and a Lactobacillus acidophilus-based direct-fed microbial, intended to control E. coli O157, had no effect on fecal shedding of O26 assessed by culture-based or PCR-based method.
Bacterial diarrheal diseases remain an important leading cause of preventable death, especially among children under five in developing countries. In the American continent, diarrheal disease and other health complications caused by Escherichia coli constitute a major public health problem, and, therefore, several research groups have dedicated their effort to understand this pathogen and provide feasible solutions to prevent, treat and reduce E. coli infections. The Latin American Coalition for Escherichia coli Research (LACER) was created as a multidisciplinary network of international research groups working with E. coli with the ultimate goal of advancing understanding of E. coli, and to prepare the next generation of American E. coli investigators. As such, this book compiles the knowledge of these investigators about E. coli, a commensal bacteria living inside its host, and a pathogen causing disease in animals and humans. Escherichia coli in the Americas contains a series of 15 chapters written by experts, covering basic concepts regarding the different categories of E. coli, including their environmental niche, virulence mechanisms, host reservoir, and disease outcomes, as well as diagnosis, vaccine development and treatment. This book's target audience include trainees and students learning about the basic and clinical aspects of E. coli pathogenesis, as well as experts around the globe who wish to learn more about this pathogen and the public health impact this bacteria has in America.
Nanobiosensors for Bio-molecular Targeting presents the latest analytical methods for the detection of different substances in the range of small molecules to whole cells, exploring the advantages and disadvantages of each method. Biosensors combine the component of biological origin and physicochemical detector to show the presence of analytes in a given sample. The use of bionanotechnology has led to a significant advancement in the progression of nanobiosensors and has been effectively used for biomedical diagnosis. Explains the detection techniques used by nanosensors, exploring the strengths and weaknesses of each for the detection of disease Shows how biosensors are used to detect various types of biomolecules Demonstrates how the use of nanomaterials makes biosensors both cheaper and more efficient
Strains of pathogenic Escherichia coli that are characterized by their ability to produce Shiga toxins are referred to as Shiga toxin-producing E. coli (STEC). STEC are an important cause of foodborne disease and infections have been associated with a wide range of human clinical illnesses ranging from mild non-bloody diarrhoea to bloody diarrhoea (BD) and haemolytic uraemic syndrome (HUS) which often includes kidney failure. A high proportion of patients are hospitalized, some develop end-stage renal disease (ESRD) and some die. The information requested by CCFH is divided into three main areas: the global burden of disease and source attribution; hazard identification and characterization; and monitoring, including the status of the currently available analytical methods. This report provides an overview of the work undertaken in response to the request from the CCFH and provides the conclusions and advice of the Expert Group based on the currently available information.