Shiga Toxin Research Articles

Immunomodulatory effect of aqueous extract of Termitomyces striatus mushroom on enterohemorrhagic Escherichia coli-infected mice

Enterohemorrhagic Escherichia coli (EHEC) generates Shiga toxins that causes immunosuppression and diarrhea. Synthetic immunomodulatory drugs are associated with numerous severe effects, necessitating the need for substitute therapeutic agents. This study aimed to determine the immunomodulatory activity of aqueous extract of Termitomyces striatus in in mice infected with EHEC. Mice in the positive control, negative control and two extract-treated groups were administered orally with 200µL of 9×108 CFU/mL EHEC. From day 5, the positive control, negative control, and extract-treated mice were orally administered with levamisole (50mg/kg body weight (bw)), phosphate-buffered saline (vehicle), and aqueous extract of Termitomyces striatus (200 and 400mg/kg body weight), respectively. The control mice received the vehicle only. After the experiment, the mice were euthanized, blood was collected, and white blood cells (WBCs) and differentials analyzed using a hematological analyzer. Immunoglobulin G (IgG), interleukin-4 (IL-4) and interferon-gamma (INF-γ) levels were also quantified using enzyme-linked immunosorbent assays. The extract significantly reduced elevated levels of WBCs, neutrophils, monocytes, eosinophils and basophils, as well as significantly enhanced the levels of lymphocytes in EHEC-infected mice (p<0.05). Besides, the extract significantly lowered the levels of INF-γ while significantly increasing the levels of IL-4 and IgG in EHEC-infected mice. Flavonoids, alkaloids, saponins, steroids and phenolics are some of the phytochemicals that were detected in T. striatus. Together, these finding revealed that T. striatus extract modulated immune response in EHEC- challenged mice by lowering leucocyte counts and INF-γ, enhancing lymphocytes, IL-4 and IgG, suggesting that the extract could be developed as an immunomodulatory agent.

Microbiological evaluation of cattle carcasses in some local slaughterhouses at Menoufia governorate

Beef carcass evaluation is generally the basis for judging the commercial value of the livestock and is consequently one of the most common quality control tests carried out in the meat industry. In the current study, sixty swabs were taken from the outer surface of neck and groin (30 of each) of freshly dressed beef carcasses taken from three local slaughterhouses located in El-Bagour, El-May and El-Shohada provinces of Menoufia governorate, Egypt. Swabs were microbiologically examined; the results revealed that groin samples had significantly (P≤0.05) lower contamination levels than that of neck samples. In addition, samples that were collected from El-May’s abattoir revealed the highest bacterial counts, followed by El-Bagour and El-Shohada, respectively. Regarding to detection of some pathogenic bacteria in the examined swab samples, S. aureus was the most detected bacteria, followed by Salmonella spp. and E. coli with total incidence 23.3, 18.3 and 15.0, respectively. However, equal number of S. aureus isolates was obtained from neck and groin samples, groin samples showed higher incidence of Salmonella and E. coli than those of neck samples. In addition, staphylococcal enterotoxins were investigated in the contaminated samples. Results revealed that all of the examined samples had enterotoxin type A only. Moreover, E. coli isolates were serotyped into O103:H2, O146:H21 and O117:H4; out of which, three strains were molecularly examined for their Shiga toxin types 1 (stx-1) and 2 (stx-2) genes; where, stx-1 was detected in 66.7% of the examined strains, while stx-2 was not detected in any of the examined isolates. On the other hand, Salmonella strains were serotyped into S. Enteritidis, S. Montevideo, S. Tsevie, S. Typhimurium and S. Shangani were the detected serotypes. Salmonella Typhimurium was detected in higher incidence than the other detected strains. So, regular monitoring of beef carcasses microbiological evaluation, and application of strict hygienic measures are recommended.

Pathogenic Factors and Recent Study on the Rapid Detection of Shiga Toxin-Producing Escherichia coli (STEC).

This comprehensive review delves into the pathogenicity and detection of Shiga Toxin-Producing Escherichia coli (STEC), shedding light on its various genetic and clinical manifestations. STEC originating from E. coli acquires pathogenicity through mobility and genetic elements. The pathogenicity of STEC is explored in terms of clinical progression, complications, and key toxins such as Shiga toxin (Stx). Stx1 and Stx2 are two distinct Stx types exhibiting different toxicities, with Stx2 often associated with severe diseases. This review also delves into Subtilase cytotoxin, an additional cytotoxin produced by some STEC strains. Pathogenic mechanisms of STEC, such as attaching and effacing intestinal lesions, are discussed, with a focus on roles of genetic factors. Plasmids in STEC can confer unique pathogenicity. Hybridization with other pathogenic E. coli can create more lethal pathogens. This review covers a range of detection methods, ranging from DNA amplification to antigen detection techniques, emphasizing the need for innovative approaches to improve the sensitivity and speed of STEC diagnosis. In conclusion, understanding diverse aspects of STEC pathogenicity and exploring enhanced diagnostic methods are critical to addressing this foodborne pathogen effectively. Pathology of Shiga toxin toxicity. STEC-derived Shiga toxin consists of one A subunit and five B subunits. Pathological symptoms of the disease can progress to HUS within two weeks after the onset of diarrhea. Shiga toxin intoxication is also associated with many complications, such as neurological and cardiac complications. This figure was reconstructed based on data from Bruyand et al.

(Invited) Pathogen Agnostic Biodetection at the Point of Need

Emerging pathogens are a constant threat to global health and human life, as exemplified by the recent COVID-19 pandemic. The availability of rapid testing - including in home tests - was instrumental in facilitating return to our 'normal' way of life. Diagnostics - especially those that can quickly and effectively inform on the condition at the point of need, enhance situational awareness and guide decision making and mitigation strategies can greatly impact our ability to counter emerging threats, epidemics and pandemics. Yet, current diagnostics are largely targeted- meaning they are tailored to identify a limited repertoire of anticipated threats. For true preparedness, it is important that the strategies we develop are pathogen agnostic and can be readily used against any organism quickly. Our team has sought inspiration from the human innate immune system in order to develop truly agnostic diagnostics. Innate immunity is medicated by pattern recognition receptors that have evolved to recognize evolutionarily conserved signatures on pathogens - allowing for their early identification. These germline receptors bind with conserved pathogen associated molecular patterns – PAMPs – produced by all pathogens, and result in the development of an associated cytokine and chemokine response. Innate immunity is thus a universal biosensor. Drawing inspiration from this natural system, our team has developed diagnostic assays for all pathogens. This involved understanding of common mechanisms of host-pathogen interactions, which then lead to the design of curated novel assay modalities - membrane insertion and lipoprotein capture - for the detection of pathogen. Beyond assay development and characterization of the response, we have evaluated the feasibility of these methods to detect signatures in blinded clinical samples for a variety of diseases (tuberculosis, invasive Salmonella, Shiga toxin carrying E. coli, Staphylococcal bacteremia, and others) with excellent sensitivity as compared to benchmark assays. These outcomes and strategies for pathogen agnostic detection will be presented.Transitioning this platform for field use, our team engineered a portable waveguide-based optical biosensor platform with integrated microfluidics for sample processes. The platform named Portable EnGineered Analytical Sensor with aUtomated Sampling (PEGASUS, R&D100 2021). This platform uses single mode planar optical waveguides as the sensing element, and builds on a bench top instrument that has been developed at the Los Alamos National Laboratory. PEGASUS miniaturizes this benchtop sensor, with integrated sample processing, moving us toward the goal of developing a truly fieldable biosensor. PEGASUS uses a different, smaller waveguide mounting apparatus, miniaturized robust components, sensing hardware and software.PEGASUS integrates Catch-all, the first ever sample processing system capable of lipid separation with ultra-sensitive optical detection of complex bioanalytes at the point of need. a microfluidic device capable of centrifugal separation of serum from blood at the point of need with a system that is compatible with biomarkers that are both hydrophilic and hydrophobic. The cross-flow filtration device separates serum from blood as efficiently as traditional methods and retains amphiphilic biomarkers in serum for detection.Whereas the scope of targets is extremely broad, the platform still relies on the use of reagents, which complicates field use in some instances. Therefore, we are beginning to explore multi-wavelength spectroscopy as a tool for the measurement of complex clinical signatures - within seconds, without reagents. Preliminary data from this development will also be presented.

A prophage encoded ribosomal RNA methyltransferase regulates the virulence of Shiga-toxin-producing Escherichia coli (STEC).

Shiga toxin (Stx) released by Shiga toxin producing Escherichia coli (STEC) causes life-threatening illness. Its production and release require induction of Stx-encoding prophage resident within the STEC genome. We identified two different STEC strains, PA2 and PA8, bearing Stx-encoding prophage whose sequences primarily differ by the position of an IS629 insertion element, yet differ in their abilities to kill eukaryotic cells and whose prophages differ in their spontaneous induction frequencies. The IS629 element in ϕPA2, disrupts an ORF predicted to encode a DNA adenine methyltransferase, whereas in ϕPA8, this element lies in an intergenic region. Introducing a plasmid expressing the methyltransferase gene product into ϕPA2 bearing-strains increases both the prophage spontaneous induction frequency and virulence to those exhibited by ϕPA8 bearing-strains. However, a plasmid bearing mutations predicted to disrupt the putative active site of the methyltransferase does not complement either of these defects. When complexed with a second protein, the methyltransferase holoenzyme preferentially uses 16S rRNA as a substrate. The second subunit is responsible for directing the preferential methylation of rRNA. Together these findings reveal a previously unrecognized role for rRNA methylation in regulating induction of Stx-encoding prophage.

Characterization of the novel temperate Escherichia coli phage phiStx2k.

A novel temperate phage, phiStx2k, was induced from a clinical Escherichia coli isolate producing Shiga toxin (Stx) 2k. The phage particles have an icosahedral head (50 nm in diameter) and a long non-contractile tail (149 nm long). The phage genome consists of 46,647 bp of double-stranded DNA with an average G + C content of 51%. Genome sequence comparisons suggested that phiStx2k represents a new genus in the class Caudoviricetes. phiStx2k was capable of converting non-Stx-producing E. coli strains to Stx producers. These results expand our knowledge on the characteristics of Stx phages and highlight the potential risks of the emergence of Stx-producing strains or novel pathogens via horizontal gene transfer.

Progression of renal damage and tubular regeneration in pregnant and non-pregnant adult female rats inoculated with a sublethal dose of Shiga toxin 2

BackgroundShiga toxin-producing Escherichia coli is the main cause of post-diarrheal hemolytic uremic syndrome (HUS) which produces acute kidney injury mainly in children, although it can also affect adults. The kidneys are the organs most affected by Shiga toxin type 2 (Stx2) in patients with HUS. However, previous studies in pregnant rats showed that a sublethal dose of Stx2 causes severe damage in the uteroplacental unit and induces abortion, whereas produces mild to moderate renal damage. The aim of the present work was to study the progression of renal injury caused by a sublethal dose of Stx2, as well as renal recovery, in pregnant and non-pregnant rats, and to investigate whether pregnancy physiology may affect renal damage progression mediated by Stx2. MethodsRenal function and histopathology was evaluated in pregnant rats intraperitoneally injected with a sublethal dose of Stx2 (0.5 ng/g bwt) at the early stage of gestation (day 8 of gestation), and results in these rats were compared over time with those observed in non-pregnant female rats injected with the same Stx2 dose. Hence, progression of cell proliferation and dedifferentiation in renal tubular epithelia was also investigated. ResultsThe sublethal dose of Stx2 induced abortion in pregnant rats as well as a significant more extended functional and histological renal injury in non-pregnant rats than in pregnant rats. Stx2 also caused decreased ability to concentrate urine in non-pregnant rats compared to their controls. However, renal water handling in pregnant rats was not altered by Stx2, and was significantly different than in non-pregnant rats. The greatest renal injury in both pregnant and non-pregnant rats was observed at 4 days post-Stx2 injection, and coincided with a significant increase in tubular epithelial proliferation. Expression of mesenchymal marker vimentin in tubular epithelia was consistent with the level of tubular damage, being higher in non-pregnant rats than in pregnant rats. Recovery from Stx2-induced kidney injury was faster in pregnant rats than in non-pregnant rats. ConclusionsAdaptive mechanisms developed during pregnancy such as changes in water handle and renal hemodynamic may contribute to lessen the Stx2-induced renal injury, perhaps at the expense of fetal loss.

Detection of Cleaved Stx2a in the Blood of STEC-Infected Patients.

Typical hemolytic uremic syndrome (HUS) is mainly caused by Shiga toxin-producing Escherichia coli (STEC) releasing Shiga toxin 2 (Stx2). Two different structures of this AB5 toxin have been described: uncleaved, with intact B and A chains, and cleaved, with intact B and a nicked A chain consisting of two fragments, A1 and A2, connected by a disulfide bond. Despite having the same toxic effect on sensitive cells, the two forms differ in their binding properties for circulating cells, serum components and complement factors, thus contributing to the pathogenesis of HUS differently. The outcome of STEC infections and the development of HUS could be influenced by the relative amounts of uncleaved or cleaved Stx2 circulating in patients' blood. Cleaved Stx2 was identified and quantified for the first time in four out of eight STEC-infected patients' sera by a method based on the inhibition of cell-free translation. Cleaved Stx2 was present in the sera of patients with toxins bound to neutrophils and in two out of three patients developing HUS, suggesting its involvement in HUS pathogenesis, although in association with other bacterial or host factors.

Retraction: Dexamethasone Rescues Neurovascular Unit Integrity from Cell Damage Caused by Systemic Administration of Shiga Toxin 2 and Lipopolysaccharide in Mice Motor Cortex

Retraction: Dexamethasone Rescues Neurovascular Unit Integrity from Cell Damage Caused by Systemic Administration of Shiga Toxin 2 and Lipopolysaccharide in Mice Motor Cortex

Antimicrobial growth promoters approved in food-producing animals in South Africa induce shiga toxin-converting bacteriophages from Escherichia coli O157:H7.

In this study, four antimicrobial growth promoters, including virginiamycin, josamycin, flavophospholipol, poly 2-propenal 2-propenoic acid and ultraviolet light, were tested for their capacity to induce stx-bacteriophages in 47 Shiga toxin-producing E. coli O157:H7 isolates. Induced bacteriophages were characterized for shiga toxin subtypes and structural genes by PCR, DNA restriction fragment length polymorphisms (RFLP) and morphological features by electron microscopy. Bacteriophages were induced from 72.3% (34/47) of the STEC O157:H7 isolates tested. Bacteriophage induction rates per induction method were as follows: ultraviolet light, 53.2% (25/47); poly 2-propenal 2-propenoic acid, 42.6% (20/47); virginiamycin, 34.0% (16/47); josamycin, 34.0% (16/47); and flavophospholipol, 29.8% (14/47). A total of 98 bacteriophages were isolated, but only 59 were digestible by NdeI, revealing 40 RFLP profiles which could be subdivided in 12 phylogenetic subgroups. Among the 98 bacteriophages, stx2a, stx2c and stx2d were present in 85.7%, 94.9% and 36.7% of bacteriophages, respectively. The Q, P, CIII, N1, N2 and IS1203 genes were found in 96.9%, 82.7%, 69.4%, 40.8%, 60.2% and 73.5% of the samples, respectively. Electron microscopy revealed four main representative morphologies which included three bacteriophages which all had long tails but different head morphologies: long hexagonal head, oval/oblong head and oval/circular head, and one bacteriophage with an icosahedral/hexagonal head with a short thick contractile tail. This study demonstrated that virginiamycin, josamycin, flavophospholipol and poly 2-propenal 2-propenoic acid induce genetically and morphologically diverse free stx-converting bacteriophages from STEC O157:H7. The possibility that these antimicrobial growth promoters may induce bacteriophages in vivo in animals and human hosts is a public health concern. Policies aimed at minimizing or banning the use of antimicrobial growth promoters should be promoted and implemented in countries where these compounds are still in use in animal agriculture.

A novel Shiga toxin 2a neutralizing antibody therapeutic with low immunogenicity and high efficacy.

Shiga toxin-producing Escherichia coli infections are difficult to treat due to the risk of antibiotic-induced stress upregulating the production of toxins, medical treatment is consequently limited to supportive care to prevent the development of hemolytic uremic syndrome (HUS). Here, we introduce a potentially therapeutic humanized mouse monoclonal antibody (Hu-mAb 2-5) targeting Stx2a, the most common Shiga toxin subtype identified from outbreaks. We demonstrate that Hu-mAb 2-5 has low immunogenicity in healthy adults ex vivo and high neutralizing efficacy in vivo, protecting mice from mortality and HUS-related tissue damage.

First report of the prevalence of Shiga toxinproducing Escherichia coli in ground beef in Quindío, Colombia.

Shiga toxin-producing Escherichia coli (STEC) is a foodborne pathogen associated with clinical cases of diarrhea in humans. Its main virulence factors are the Shiga toxins (Stx1 and Stx2). Cattle are the main reservoir of STEC, and many outbreaks in humans have been related to the consumption of undercooked ground beef contaminated with this pathogen. To determine the prevalence of STEC in ground beef commercialized in all the butcher shops of a township in the department of Quindío and to characterize the virulence genes of the strains found. Thirty ground beef samples were taken in three different times; stx genes and other STEC virulence factors (eae, ehxA, saa) were detected by multiplex PCR. The overall prevalence of STEC was 33.33 % (10/30 positive samples). We isolated eight non-O157 (LEE-negative) strains with four different genetic profiles: stx2 / stx2-ehxA-saa / stx1-stx2-ehxA-saa / stx1-saa. This is the first report on the prevalence of STEC in ground beef in a township in the department of Quindío.

Compendium of Measures to Prevent Disease Associated with Animals in Public Settings, 2023

Abstract Opportunities for the public to interact with animals in public settings such as petting zoos, fairs, and farm visits can be valuable learning and entertainment experiences. However, zoonotic disease transmission from healthy animals on exhibit and their environments, as well as injuries and other health problems, may result from these interactions if steps are not taken to minimize risks. The 2023 Compendium of Measures to Prevent Disease Associated with Animals in Public Settings provides background about these potential risks and updates recommendations for reducing those risks. Enteric zoonotic disease risks from animal contact in public settings include Shiga toxin–producing Escherichia coli infections, cryptosporidiosis, and campylobacteriosis. These infections occur when pathogens are accidentally ingested by individuals following direct contact with animals or contaminated objects. Zoonotic influenza infections can occur in a similar manner or through aerosols generated from infected animals. Other infectious diseases, parasitic infections, bites, and other injuries are also possible hazards in these settings. Inadequate handwashing and facility design flaws are frequent contributors to these risks, especially on farms or temporary exhibits not specifically designed for public use. Venues should provide sufficient handwashing stations and establish clearly defined animal contact areas. Operators should educate visitors about potential disease and injury risks, steps necessary to minimize exposure, and the importance of handwashing after the visit. Additionally, operators and visitors alike should realize that certain individuals are at heightened risk for zoonotic infections. Signs, handouts, and other educational materials are publicly available in a toolkit that accompanies the compendium.

Enteric pharmacokinetics of monomeric and multimeric camelid nanobody single-domain antibodies.

Single-domain antibodies (sdAbs) derived from Camelidae heavy-chain-only antibodies (also called nanobodies or VHHs) have advantages over conventional antibodies in terms of their small size and stability to pH and temperature extremes, their ability to express well in microbial hosts, and to be functionally multimerized for enhanced properties. For these reasons, VHHs are showing promise as enteric disease therapeutics, yet little is known as to their pharmacokinetics (PK) within the digestive tract. To improve understanding of enteric VHH PK, we investigated the functional and structural stability of monomeric and multimeric camelid VHH-agents following in vitro incubation with intestinal extracts (chyme) from rabbits and pigs or fecal extracts from human sources, and in vivo in rabbits. The results showed that unstructured domains such as epitopic tags and flexible spacers composed of different amino acid sequences were rapidly degraded by enteric proteases while the functional core VHHs were much more stable to these treatments. Individual VHHs were widely variable in their functional stability to GI tract proteases. Some VHH-based agents which neutralize enteric Shiga toxin Stx2 displayed a functional stability to chyme incubations comparable to that of Stx2-neutralizing IgG and IgA mAbs, thus indicating that selected nanobodies can approach the functional stability of conventional immunoglobulins. Enteric PK data obtained from in vitro incubation studies were consistent with similar incubations performed in vivo in rabbit surgical gut loops. These findings have broad implications for enteric use of VHH-based agents, particularly VHH fusion proteins.

Genetic Characterization of Intimin Gene (eae) in Clinical Shiga Toxin-Producing Escherichia coli Strains from Pediatric Patients in Finland.

Shiga toxin (Stx)-producing Escherichia coli (STEC) infections cause outbreaks of severe disease in children ranging from bloody diarrhea to hemolytic uremic syndrome (HUS). The adherent factor intimin, encoded by eae, can facilitate the colonization process of strains and is frequently associated with severe disease. The purpose of this study was to examine and analyze the prevalence and polymorphisms of eae in clinical STEC strains from pediatric patients under 17 years old with and without HUS, and to assess the pathogenic risk of different eae subtypes. We studied 240 STEC strains isolated from pediatric patients in Finland with whole genome sequencing. The gene eae was present in 209 (87.1%) strains, among which 49 (23.4%) were from patients with HUS, and 160 (76.6%) were from patients without HUS. O157:H7 (126, 60.3%) was the most predominant serotype among eae-positive STEC strains. Twenty-three different eae genotypes were identified, which were categorized into five eae subtypes, i.e., γ1, β3, ε1, θ and ζ3. The subtype eae-γ1 was significantly overrepresented in strains from patients aged 5-17 years, while β3 and ε1 were more commonly found in strains from patients under 5 years. All O157:H7 strains carried eae-γ1; among non-O157 strains, strains of each serotype harbored one eae subtype. No association was observed between the presence of eae/its subtypes and HUS. However, the combination of eae-γ1+stx2a was significantly associated with HUS. In conclusion, this study demonstrated a high occurrence and genetic variety of eae in clinical STEC from pediatric patients under 17 years old in Finland, and that eae is not essential for STEC-associated HUS. However, the combination of certain eae subtypes with stx subtypes, i.e., eae-γ1+stx2a, may be used as risk predictors for the development of severe disease in children.

A Review on Ecoli 0157: H7 (EHEC)

Escherichia coli O157:H7 is an important food borne and water borne pathogen is first isolated in 1982, shiga toxin. producing Ecoli O157:H7 which causes diarrhea, hemorrhagic colitis and haemolytic uremic Syndrome in humans. Haemolytic uremic. Syndrome (Hus) is a Serious condition and upto 50% of Hus patients can develop renal acute dysfunction or blood pressure. Ecoli 0157:H7 infects the alimentary tract and induces abdominal cramps with hemorrhagic diarrhea and it can be transmitted by person to person through fecal shedding and 11%. of infection are estimated. Enterotoxigenic Escherichia coli(ETEC) strain are the main cause of the diarrheal disease problems. ETEC strains are the bacteria which cause the acute watery diarrhea in chinldren. Diarrheagenic Ecoli infection are more common in Africa countries. The occurance of antimicrobial resistance depends upon the type of antimicrobialagent,bacterial strain,dose,time and mode of ministration. some of Ecoli 0517:H7 occurs after taking the undercooked meat or deficiency pasteurized dairy products. And some of Ecoli 0157:H7 occurs due ro exposal to contaminated water from potable drinking sources,pools and lakes and foods such as inappropriate washed leafy green vegetables and fruits,apple juice and direct contact with contaminated animals. Ecoli 0157:H7 shiga toxin causes leukocyte aggregation,platelet aggregation,microthrombin formation, apoptosis of affected cells ,renal dysfunction. shiga toxin not only effect kidney but it can also effect in diffuse vasculitic injuries which cause the effect on multiple organ system and organ failure.

Gb3-Coated Bovine Milk Exosomes as a Practical Neutralizer for Shiga Toxin.

Shiga toxin (Stx) is associated with foodborne infections of some Shigella spp. and Shiga toxin-producing Escherichia coli (STEC), leading to life-threatening hemolytic uremic syndrome (HUS). Target-specific therapeutics against HUS are currently unavailable in clinical practice. Herein, we reported the construction and in vitro characterization of Gb3-coated bovine milk exosomes (Gb3-mExo) as a multivalent Shiga toxin neutralizer, utilizing the natural advantages of milk exosomes (mExo) in drug delivery and multivalent interactions between Stx and its receptor Gb3. Gb3-mExo constructs were achieved by conjugating mExo with the Gb3 derivatives containing stearic acid-derived lipid tail, which was prepared through an efficient chemoenzymatic approach. The constructs were able to potently neutralize the binding of the B subunit of Stx2 (Stx2B) to receptor Gb3 immobilized on the plate or expressed on model cells. General safety of the constructs was evidenced by the cytotoxicity analysis and hemolysis assay. In addition to the excellent stability under conventional storage and handling conditions, the construct can also retain most of its neutralization potency under gastrointestinal pH extremes, showing the potential for oral administration. Considering the natural availability and excellent biocompatibility of mExo, Gb3-mExo conjugates should prove to be a practical prophylactic and therapeutic for the Shiga toxin-related infections.

A Moderate Decrease in ADAMTS13 Activity Correlates with the Severity of STEC-HUS.

Atypical hemolytic uremic syndrome (HUS) develops as a result of damage to the endothelium of microvasculature vessels by Shiga toxin produced by enterohemorrhagic Escherichia coli (STEC-HUS). STEC-HUS remains the leading cause of acute kidney injury (AKI) in children aged 6 months to 5 years. The pathomorphological essence of the disease is the development of thrombotic microangiopathy (TMA). One of the key causes of TMA is an imbalance in the ADAMTS13-von Willebrand factor (vWF)-platelet system. The goal of the work was to clarify the role of a moderate decrease in ADAMTS13 activity in the pathogenesis of STEC-HUS. The activity of ADAMTS13 was determined in 138 children (4 months-14.7 years) in the acute period of STEC-HUS and the features of the course of the disease in these patients were analyzed. The study revealed a decrease in the activity and concentration of ADAMTS13 in 79.8% and 90.6% of patients, respectively. Measurements of von Willebrand factor antigen content and the activity of von Willebrand factor in the blood plasma of part of these patients were carried out. In 48.6% and 34.4% of cases, there was an increase in the antigen concentration and the activity of the Willebrand factor, respectively. Thrombocytopenia was diagnosed in 97.8% of children. We have demonstrated that moderately reduced ADAMTS13 activity correlates with the risk of severe manifestations of STEC-HUS in children; the rate of developing multiple organ failure, cerebral disorders, pulmonary edema, and acute kidney injury with the need for dialysis increases. It is assumed that reduction in ADAMTS13 activity may serve as a predictor of disease severity.

Genomic and Phenotypic Characterization of Shiga Toxin-Producing Escherichia albertii Strains Isolated from Wild Birds in a Major Agricultural Region in California.

Escherichia albertii is an emerging foodborne pathogen. To better understand the pathogenesis and health risk of this pathogen, comparative genomics and phenotypic characterization were applied to assess the pathogenicity potential of E. albertii strains isolated from wild birds in a major agricultural region in California. Shiga toxin genes stx2f were present in all avian strains. Pangenome analyses of 20 complete genomes revealed a total of 11,249 genes, of which nearly 80% were accessory genes. Both core gene-based phylogenetic and accessory gene-based relatedness analyses consistently grouped the three stx2f-positive clinical strains with the five avian strains carrying ST7971. Among the three Stx2f-converting prophage integration sites identified, ssrA was the most common one. Besides the locus of enterocyte effacement and type three secretion system, the high pathogenicity island, OI-122, and type six secretion systems were identified. Substantial strain variation in virulence gene repertoire, Shiga toxin production, and cytotoxicity were revealed. Six avian strains exhibited significantly higher cytotoxicity than that of stx2f-positive E. coli, and three of them exhibited a comparable level of cytotoxicity with that of enterohemorrhagic E. coli outbreak strains, suggesting that wild birds could serve as a reservoir of E. albertii strains with great potential to cause severe diseases in humans.

Detection and characterization of circulating microvesicles containing Shiga toxin type 2 in a rat model of Hemolytic Uremic Syndrome

Shiga toxin producing Escherichia coli (STEC) are foodborne pathogens that release Shiga toxin (Stx), virulence factor responsible for the development of Hemolytic Uremic Syndrome (HUS). Stx causes endothelial cell damage, which leads to platelets deposition and thrombi formation within the microvasculature. It has been described that Stx activates blood cells and induces the shedding of proinflammatory and prothrombotic microvesicles (MVs) containing the toxin. In this sense, it has been postulated that MVs containing Stx2 (MVs-Stx2+) can contribute to the physiopathology of HUS, allowing Stx2 to reach the target organs while evading the immune system. In this work, we propose that circulating MVs-Stx2+ can be a potential biomarker for the diagnosis and prognosis of STEC infections and HUS progression. We developed a rat HUS model by the intraperitoneal injection of a sublethal dose of Stx2 and observed: decrease in body weight, increase of creatinine and urea levels, decrease of creatinine clearance and histological renal damages. After characterization of renal damages, we investigated circulating total MVs and MVs-Stx2+ by flow cytometry at different times after Stx2 injection. Additionally, we evaluated the correlation of biochemical parameters such as creatinine and urea in plasma with MVs-Stx2+. As a result, we found a significant circulation of MVs-Stx2+ at 72 and 96 h after Stx2 injection, nevertheless no correlation with creatinine and urea plasma levels were detected. Our results suggest that MVs-Stx2+ may be an additional biomarker for the characterization and diagnosis of HUS progression. A further analysis is required in order to validate MVs-Stx2+ as biomarker of the disease.