Coli O157 Research Articles

Conductometric immunosensor for specific Escherichia coli O157:H7 detection on chemically funcationalizaed interdigitated aptasensor

Escherichia coli O157:H7 is a strain of Escherichia coli known for causing foodborne illness through the consumption of contaminated or raw food. To detect this pathogen, a conductometric immunosensor was developed using a conductometric sensing approach. The sensor was constructed on an interdigitated electrode and modified with a monoclonal anti-Escherichia coli O157:H7 aptamer. A total of 200 electrode pairs were fabricated and modified to bind to the target molecule replica. The binding replica, acting as the bio-recognizer, was linked to the electrode surface using 3-Aminopropyl triethoxysilane. The sensor exhibited excellent performance, detecting Escherichia coli O157:H7 in a short time frame and demonstrating a wide detection range of 1 fM to 1 nM. Concentrations of Escherichia coli O157:H7 were detected within this range, with a minimum detection limit of 1 fM. This innovative sensor offers simplicity, speed, high sensitivity, selectivity, and the potential for rapid sample processing. The potential of this proposed biosensor is particularly beneficial in applications such as drug screening, environmental monitoring, and disease diagnosis, where real-time information on biomolecular interactions is crucial for timely decision-making and where cross-reactivity or interference may compromise the accuracy of the analysis.

Comparison of Three Preharvest Sampling Strategies to Monitor Pathogens in Cattle Lairage Areas

The objective of this study was to compare preharvest monitoring strategies by evaluating three different sampling methods in the lairage area to determine pathogen recovery for each sampling method and incoming pathogen prevalence from the cattle to inform in-plant decision making. Samples were gathered over a 5-month period, from February to June 2022, at a harvesting and processing facility located in Eastern Nebraska. Sampling methods included (i) fecal pats, (ii) boot swabs, and (iii) MicroTally swab. A total of 329 samples were collected over the study period (fecal pats: n = 105, boot swabs: n = 104, and MicroTally swabs: n = 120). Specific media combinations, an incubation temperature of 42°C, and incubation timepoints (18–24 h) were utilized for each matrix and the prevalence of Salmonella, Escherichia coli O157:H7, and six non-O157 Shiga-toxin producing E. coli (STEC) was evaluated using the BAX system Real-Time PCR assay. Overall, results from the study concluded that boot swabs were an effective sampling method for pathogen detection in the cattle lairage area. Boot swabs (97.1%) were statistically more likely to detect for Salmonella (p < 0.05) when compared to fecal pats (67.6%) and MicroTally swab (77.5%) methods. For E. coli O157:H7 and STEC – O26, O121, O45, and O103 prevalence, boot swabs were significantly better at detecting for these pathogens (p < 0.05) than MicroTally swabs (OR = 3.16 – 11.95) and a comparable sampling method to fecal pats (OR = 0.93 – 2.01, p > 0.05). Lastly, all three sampling methods detected a very low prevalence for E. coli O111 and O145; therefore, no further analysis was conducted. The boot swab sampling method was strongly favored because they require little training to implement, are inexpensive, and they do not require much sampling labor; therefore, would be a simple and effective sampling method to implement within the industry to evaluate pathogen prevalence preharvest.

In vitro and In silico Evaluation of Antibacterial Effects in Some Oxadiazole Compounds against Escherichia coli O157: H7

Background: Escherichia coli O157:H7 is a significant pathogen responsible for causing diarrhea in humans. Oxadiazoles are known for their diverse range of biological activities. Objectives: This study aimed to assess the anti-E. coli O157:H7 effects of 1, 3, and 4-oxadiazole derivatives. Methods: Compounds containing an oxadiazole central core were synthesized anew. In vitro assays, including agar well diffusion, minimum inhibitory concentration, and minimum bactericidal concentration, were conducted. The molecular structures of the oxadiazole derivatives were optimized using the mm2 methodology with Chem3D v20.1.1.125 software. The ligand's inhibitory potential against the active sites of stx-1 and stx-2 was assessed using Autodock Vina software. The results were analyzed using Discovery Studio v16.1.0 software. Results: The findings indicated that compound C ((2E)-3-(3,4-difluorophenyl)-2-(5-(hydroxy(pyridin-2-yl)meth yl)-1,3,4-oxadiazol-2-yl)-N-methylacrylimidic acid) exhibited more potent anti-E. coli O157:H7 effects compared to other compounds and the control sample. Furthermore, in silico results demonstrated that compound C exhibited inhibitory effects against stx-1 and stx-2 by forming hydrogen bonds for inhibition. Conclusions: Compounds featuring a fluorophenyl structure with a 1, 3, and 4-oxadiazole core have the potential to serve as anti-E. coli O157:H7 agents for the development of therapeutic drugs.

Silver Nanoparticle-Aminogylcosides Conjugation for Enhanced Control of Pathogenic E. Coli O157:H7

Silver Nanoparticle-Aminogylcosides Conjugation for Enhanced Control of Pathogenic E. Coli O157:H7

Cultivar was more influential than bacterial strain and other experimental factors in recovery of Escherichia coli O157:H7 populations from inoculated live Romaine lettuce plants.

The varied choice of bacterial strain, plant cultivar, and method used to inoculate, retrieve, and enumerate Escherichia coli O157:H7 from live plants could affect comparability among studies evaluating lettuce-enterobacterial interactions. Cultivar, bacterial strain, incubation time, leaf side inoculated, and sample processing method were assessed for their influence in recovering and quantifying E. coli O157:H7 from live Romaine lettuce. Cultivar exerted the strongest effect on E. coli O157:H7 counts, which held up even when cultivar was considered in interactions with other factors. Recovery from the popularly grown green Romaine "Rio Bravo" was higher than from the red variety "Outredgeous." Other modulating variables were incubation time, strain, and leaf side inoculated. Sample processing method was not significant. Incubation for 24 hours post-lettuce inoculation yielded greater counts than 48 hours, but was affected by lettuce cultivar, bacterial strain, and leaf side inoculated. Higher counts obtained for strain EDL933 compared to a lettuce outbreak strain 2705C emphasized the importance of selecting relevant strains for the system being studied. Inoculating the abaxial side of leaves gave higher counts than adaxial surface inoculation, although this factor interacted with strain and incubation period. Our findings highlight the importance of studying interactions between appropriate bacterial strains and plant cultivars for more relevant research results, and of standardizing inoculation and incubation procedures. The strong effect of cultivar exerted on the E. coli O157:H7-lettuce association supports the need to start reporting cultivar information for illness outbreaks to facilitate the identification and study of plant traits that impact food safety risk.IMPORTANCEThe contamination of Romaine lettuce with Escherichia coli O157:H7 has been linked to multiple foodborne disease outbreaks, but variability in the methods used to evaluate E. coli O157:H7 association with live lettuce plants complicates the comparability of different studies. In this study, various experimental variables and sample processing methods for recovering and quantifying E. coli O157:H7 from live Romaine lettuce were assessed. Cultivar was found to exert the strongest influence on E. coli O157:H7 retrieval from lettuce. Other modulating factors were bacterial incubation time on plants, strain, and leaf side inoculated, while sample processing method had no impact. Our findings highlight the importance of selecting relevant cultivars and strains, and of standardizing inoculation and incubation procedures, in these types of assessments. Moreover, results support the need to start reporting cultivars implicated in foodborne illness outbreaks to facilitate the identification and study of plant traits that impact food safety risk.

Carbon dots and covalent organic frameworks based FRET immunosensor for sensitive detection of Escherichia coli O157:H7

The combination of carbon dots (CDs) with covalent organic frameworks (COFs) was used to design an innovative sensor based on fluorescence resonance energy transfer (FRET) for the detection of Escherichia coli O157:H7 (E. coli O157:H7) in food samples. Carbon dots were used as fluorescence donors, covalent organic frameworks as fluorescence acceptors. The antibody (Ab) specific to E. coli O157:H7 was used to form a CD-Ab-COF immunosensor by linking CDs and COFs. The antibody was specifically bound with E. coli O157:H7, which caused the connection between CDs and COFs to be interrupted, and the carbon dots exhibited fluorescence restoration. The sensor exhibited a linear detection range spanning from 0 to 106 CFU/mL, with the limit of detection (LOD) of 7 CFU/mL. The analytical performance of the developed immunosensor was evaluated using spiked food samples with different concentrations of E. coli O157:H7, validating the capability of assessing risks in food testing.

Microbial interventions in yak colibacillosis: Lactobacillus-mediated regulation of intestinal barrier.

The etiology of Escherichia coli in yaks, along with its drug resistance, results in economic losses within the yak breeding industry. The utilization of lactic acid bacteria treatment has emerged as a viable alternative to antibiotics in managing colibacillosis. To elucidate the therapeutic mechanisms of Lactobacillus against Escherichia coli-induced intestinal barrier damage in yaks, we employed yak epithelial cells as the experimental model and established a monolayer epithelial barrier using Transwell. The study encompassed four groups: a control group, a model group (exposed to E. coli O78), a low-dose Lactobacillus group (E. coli O78 + 1 × 105CFU LAB), and a high-dose Lactobacillus group (E. coli O78 + 1 × 107CFU LAB). Various techniques, including transmembrane resistance measurement, CFU counting, RT-qPCR, and Western Blot, were employed to assess indicators related to cell barrier permeability and tight junction integrity. In the Model group, Escherichia coli O78 significantly compromised the permeability and tight junction integrity of the yak epithelial barrier. It resulted in decreased transmembrane resistance, elevated FD4 flux, and bacterial translocation. Furthermore, it downregulated the mRNA and protein expression of MUC2, Occludin, and ZO-1, while upregulating the mRNA expression and protein expression of FABP2 and Zonulin, thereby impairing intestinal barrier function. Contrastingly, Lactobacillus exhibited a remarkable protective effect. It substantially increased transmembrane resistance, mitigated FD4 flux, and reduced bacterial translocation. Moreover, it significantly upregulated the mRNA and protein expression of MUC2, Occludin, and ZO-1, while downregulating the mRNA and protein expression of FABP2 and Zonulin. Notably, high-dose LAB demonstrated superior regulatory effects compared to the low-dose LAB group. In conclusion, our findings suggest that Lactobacillus holds promise in treating yak colibacillosis by enhancing mucin and tight junction protein expression. Furthermore, we propose that Lactobacillus achieves these effects through the regulation of Zonulin.

Effect of gas ultrafine bubbles on the potency of antimicrobials against Escherichia coliO157:H7 biofilms on various food processing surfaces

AbstractThis study investigated the impact of incorporating gas [air, carbon dioxide (CO2), and nitrogen (N2)] UFB on the potency of chlorine (Cl2; 50, 100, and 200 ppm) and peracetic acid (PAA; 20, 40, and 80 ppm) antimicrobial (AM) solutions against fresh (3 days) and aged (30 days) E. coli O157:H7 biofilms on polypropylene, silicone, and stainless‐steel surfaces. The biofilms were statically grown on polypropylene, silicone, and stainless‐steel coupons (7.62 × 2.54 cm) at 25°C for 3 or 30 days by immersing in a 3‐strain cocktail of E. coli. The incorporation of air, CO2, and N2 UFB in AM solutions resulted in significantly increased log reductions (2.1–3.7 logs) in fresh and aged E. coli biofilms on all surfaces compared to solutions without UFB, except for N2 UFB on aged stainless‐steel biofilms and air UFB on aged polypropylene biofilms, which resulted in similar log reductions as solutions without UFB (1.5–2.1 logs).

Minimizing Escherichia coli O157:H7 contamination in indoor farming: effects of cultivar type and ultra-violet light quality.

Bacterial contamination of produce is a concern in indoor farming due to close plant spacing, recycling irrigation, warm temperatures, and high relative humidity during production. Cultivars that inherently resist contamination and photo-sanitization using ultraviolet (UV) radiation during the production phase can reduce bacterial contamination. However, there is limited information to support their use in indoor farming. Lettuce (Lactuca sativa) cultivars with varying plant architectures grown in a custom-built indoor farm exhibited differences in E. coli O157:H7 survival after inoculation. The survival of E. coli O157:H7 was lowest in the leaf cultivar (open architecture) and highest in the romaine and oakleaf cultivars (compact architecture). Of the different UV wavelengths that were tested (UV-A, UV-A + B, UV-A + C), UV A + C at an intensity of 54.5 μmol m-2 s-1 (with 3.5 μmol m-2 s-1 of UV-C), provided for 15 min every day, was found to be most efficacious in reducing the E. coli O157:H7 survival on romaine lettuce with no negative effects on plant growth and quality. Contamination of E. coli O157:H7 on lettuce plants can be reduced and the food safety levels in indoor farms can be increased by selecting cultivars with an open leaf architecture coupled with photo-sanitization using low and frequent exposure to UV A + C radiation. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Pistacia terebinthus MEYVESİ: GIDA BOZULMASINI ÖNLEMEK İÇİN BİR ALTERNATİF

The aim of this work was to investigate the usage possibilities of P. terebinthus fruit, which has a limited usage area, in the food industry. Antimicrobial activity of hexane extract obtained from P. terebinthus fruits from Adıyaman (Turkey) was determined by disc diffusion and micro-dilution assays against test microorganisms. In addition, the antibacterial activity of the extract on Escherichia coli O157:H7, one of the most important food-borne pathogens, was determined by viable cell count using a macro-dilution assay. The potential for use of the extract with probiotic candidate lactic acid bacteria (LAB) strains was also investigated. The hexane extract presented antimicrobial activity against all the tested microorganisms, with good inhibition zone diameters between 10.51 mm and 18.02 mm. MIC and MFC or MBC values of P. terebinthus fruit extract were determined as 5-80 µg/µL against all tested microorganisms. The lowest MIC and MBC values (5 µg/µL) of the extract were obtained against E. coli O157:H7. Macro-dilution assay results indicated that the P. terebinthus extract at various concentrations (5-10-20 mg/mL) inhibited the growth of E. coli O157:H7 more than the control group after all of the incubation hours. No viable E. coli O157:H7 cells were detected after 48 hours at all concentrations. The extract showed low antimicrobial activity, and relatively high bactericidal concentration on probiotic candidate LAB strains. This shows that P. terebinthus hexane extract at appropriate concentrations can be used together with probiotic strains as a natural preservative and biopreservative to prevent food spoilage and extend shelf life.

Effect of Ground Beef Irradiation on Annual Nontyphoidal Salmonella and Escherichia coli O157 Burden and Direct Healthcare Costs in the United States: A Simulation Study

Over 20% of E. coli O157 illnesses and over 5% of Salmonella illnesses are estimated to be attributable to beef consumption in the United States. Irradiating ground beef is one possible method to reduce disease burden. We simulated the effect of ground beef irradiation on illnesses, hospitalizations, deaths, and direct healthcare costs from ground beef-associated E. coli O157 and Salmonella illnesses in the United States. To estimate the fraction of illnesses, hospitalizations, deaths, and direct healthcare costs preventable by ground beef irradiation, we multiplied the disease burden attributable to ground beef; the estimated percentage of ground beef sold that is not currently irradiated; the percentage of unirradiated ground beef that would be irradiated; and the percentage reduction in risk of illness after irradiation. We multiplied this fraction by estimates of burden and direct healthcare costs to calculate the numbers or amounts averted. Model inputs were obtained from the literature and expert opinion. We used Monte Carlo simulation to incorporate uncertainty in inputs into model estimates. Simulation outcomes were summarized with means and 95% uncertainty intervals (UI). Irradiating 50% of the currently unirradiated ground beef supply would avert 3,285 (95% UI: 624–9,977) E. coli O157 illnesses, 135 (95% UI: 24–397) hospitalizations, 197 (95% UI: 34–631) hemolytic uremic syndrome cases, 2 (95% UI: 0–16) deaths, and $2,972,656 (95% UI: $254,708–$14,496,916) in direct healthcare costs annually. For Salmonella, irradiation would avert 20,308 (95% UI: 9,858–38,903) illnesses, 400 (95% UI: 158–834) hospitalizations, 6 (95% UI: 0–18) deaths, and $7,318,632 (95% UI: $1,436,141–$26,439,493) in direct healthcare costs. Increasing ground beef irradiation could reduce E. coli O157 and Salmonella burden in the United States. Additional studies should assess whether targeted irradiation of higher-risk ground beef products could prevent similar numbers of illnesses with less total product irradiated.

Inactivation mechanism and kinetics of dielectric barrier discharge plasma against Escherichia coli O157:H7 in phosphate-buffered saline

ABSTRACT This study investigated the inactivation mechanism and kinetics of dielectric barrier discharges (DBD) plasma against Escherichia coli O157:H7. DBD plasma exhibited a remarkable bactericidal effect, and the inactivation kinetics was well described with the Weibull model. Using the ambient air as the working gas, surviving E. coli O157:H7 in phosphate-buffered saline achieved an undetectable level (<10 CFU/mL) after treatment at 18 W for 60 s. DBD plasma altered cell morphology, destructed both the outer and inner membranes, and increased intracellular reactive oxygen species levels. Moreover, DBD plasma significantly decreased the pH of the suspended medium, and increased its oxidation reduction potential and active species levels, which could be mainly responsible for microbial inactivation. The presence of glutathione or N-acetyl-L-cysteine weakened the antibacterial activity of DBD plasma due to their reactive oxygen and nitrogen species (RONS) scavenging capacity, demonstrating that RONS generated by DBD plasma played important roles in decontamination.

A new Rogue-like Escherichia phage UDF157lw to control Escherichia coli O157:H7.

Shiga toxin-producing Escherichia coli (STEC) O157:H7 is one of the notorious foodborne pathogens causing high mortality through the consumption of contaminated food items. The food safety risk from STEC pathogens could escalate when a group of bacterial cells aggregates to form a biofilm. Bacterial biofilm can diminish the effects of various antimicrobial interventions and enhance the pathogenicity of the pathogens. Therefore, there is an urgent need to have effective control measurements. Bacteriophages can kill the target bacterial cells through lytic infection, and some enzymes produced during the infection have the capability to penetrate the biofilm for mitigation compared to traditional interventions. This study aimed to characterize a new Escherichia phage vB_EcoS-UDF157lw (or UDF157lw) and determine its antimicrobial efficacy against E. coli O157:H7. Phage characterization included biological approaches, including phage morphology, one-step growth curve, stability tests (pH and temperature), and genomic approaches (whole-genome sequencing). Later, antimicrobial activity tests, including productive infection against susceptible bacterial strains, in vitro antimicrobial activity, and anti-biofilm, were conducted. UDF157lw is a new member of the phages belonging to the Rogunavirus genus, comprising a long and non-contractile tail, isolated from bovine feces and shares close genomic evolutionary similarities with Escherichia phages vB_EcoS-BECP10 and bV_EcoS_AKS96. When used against E. coli O157:H7 (ATCC35150), phage UDF157lw exhibited a latent period of 14 min and a burst size of 110 PFU per infected cell. The phage remained viable in a wide range of pH values (pH 4-11) and temperatures (4-60°C). No virulence genes, such as stx, lysogenic genes, and antibiotic resistance genes, were found. Phage UDF157lw demonstrated high infection efficiencies against different E. coli O157:H7 and generic E. coli strains. In addition, UDF157lw encoded a unique major tail protein (ORF_26) with prominent depolymerase enzyme activity against various E. coli O157:H7 strains, causing large plaque sizes. In contrast to the phage without encoding depolymerase gene, UDF157lw was able to reduce the 24-h and 48-h E. coli O157:H7 biofilm after 1-h phage treatment. The findings of this study provide insights into a new member of the Rogunavirus phages and demonstrate its antimicrobial potential against E. coli O157:H7 in vitro.

Fighting Microbial Infections from Escherichia coli O157:H7: The Combined Use of Three Essential Oils of the Cymbopogon Genus and a Derivative of Esculentin-1a Peptide.

The absence of effective therapy against Escherichia coli O157:H7 infections has led to the need to develop new antimicrobial agents. As the use of synergistic combinations of natural antimicrobial compounds is growing as a new weapon in the fight against multidrug-resistant bacteria, here, we have tested new synergistic combinations of natural agents. Notably, we investigated a possible synergistic effect of combinations of essential oils and natural peptides to counteract the formation of biofilm. We chose three essential oils (i.e., Cymbopogon citratus, C. flexuosus and C. martinii) and one peptide already studied in our previous works. We determined the fractional inhibitory concentration (FIC) by analyzing the combination of the peptide derived from esculentin-1a, Esc(1-21), with the three essential oils. We also studied the effects of combinations by time-kill curves, scanning electron microscopy on biofilm and Sytox Green on cell membrane permeability. Finally, we analyzed the expression of different genes implicated in motility, biofilm formation and stress responses. The results showed a different pattern of gene expression in bacteria treated with the mixtures compared to those treated with the peptide or the single C. citratus essential oil. In conclusion, we demonstrated that the three essential oils used in combination with the peptide showed synergy against the E. coli O157:H7, proving attractive as an alternative strategy against E. coli pathogen infections.

An Emerging Foodborne Pathogen Spotlight: A Bibliometric Analysis and Scholarly Review of Escherichia coli O157 Research.

Foodborne infections pose a substantial global threat, causing an estimated 600 million illnesses and resulting in approximately 420,000 deaths annually. Among the diverse array of pathogens implicated in these infections, Escherichia coli (E. coli), specifically the O157 strain (E. coli O157), emerges as a prominent pathogen associated with severe outbreaks. This study employs a comprehensive bibliometric analysis and scholarly review focused on E. coli O157 research. The bibliometric analysis highlights the significant role played by the United States in the E. coli O157 research domain. Further exploration underscores the noteworthy contributions of the researcher Doyle MP, whose body of work, consisting of 84 documents and an impressive H-Index of 49, reflects their substantial impact in the field. Recent research trends indicate a discernible shift towards innovative detection methods, exemplified by the adoption of CRISPR-CAS and Loop-Mediated Isothermal Amplification. Moreover, high-throughput whole-genome sequencing techniques are gaining prominence for the expeditious analysis of pathogenic E. coli strains. Scientists are increasingly exploring antimicrobial agents, including phage therapy, to address the challenges posed by antibiotic-resistant E. coli strains, thereby addressing critical concerns related to multi-drug resistance. This comprehensive analysis provides vital insights into the dynamic landscape of E. coli O157 research. It serves as a valuable resource for researchers, policymakers, and healthcare professionals dedicated to mitigating E. coli O157 outbreaks and advancing global public health strategies.

Evaluation of Peracetic Acid Treatment on Beef Trimmings and Subprimals Against Salmonella and E. Coli O157:H7 Within Regulatory Retained Water Limitations

The application of antimicrobial treatments to beef trimmings prior to grinding for the reduction of microbial contamination in ground beef has increased recently. However, raw single-ingredient meat products are not permitted by Food Safety and Inspection Services (FSIS) to retain more than 0.49% water resulting from postevisceration processing. The effectiveness of antimicrobials with the limited water retention is not well documented. The objective of this study was to determine the effectiveness of peracetic acid at varied concentrations against E. coli O157:H7 and Salmonella on the surface of beef trimmings and beef subprimals that was applied at industry operating parameters within the retained water requirement. One hundred and forty-four each of beef trimmings and subprimals were used to evaluate the effect of different concentrations of peracetic acid solution on reducing E. coli O157:H7 and Salmonella on surfaces of fresh beef within the FSIS requirement of ≤0.49% retained water from antimicrobial spray treatments using a conveyor system. A ten-strain cocktail mixture was inoculated on surfaces of fresh beef and subjected to water or four different concentrations of peracetic acid (130, 150, 200, and 400 ppm). Spray treatments with 130, 150, and 200 ppm peracetic acid reduced (P ≤ 0.05) E. coli O157:H7 and Salmonella at least 0.2 log on surfaces of beef trimmings and subprimals. Spray treatment with 400 ppm peracetic acid resulted in approximately 0.5 and 0.3 log reduction of E. coli O157:H7 and Salmonella, respectively. Results indicate that all concentrations (130–400 ppm) of peracetic acid significantly reduced E. coli O157:H7 and Salmonella on beef trimmings and subprimals compared to untreated controls. Thus, a range from 130 to 400 ppm of peracetic acid can be used during beef processing to improve the safety of beef trimmings and subprimals when weight gain is limited to ≤0.49% to meet regulatory requirements.

Red blood cell-derived arginase release in hemolytic uremic syndrome

BackgroundHemolysis is a cardinal feature of hemolytic uremic syndrome (HUS) and during hemolysis excess arginase 1 is released from red blood cells. Increased arginase activity leads to reduced L-arginine, as it is converted to urea and L-ornithine, and thereby reduced nitric oxide bioavailability, with secondary vascular injury. The objective of this study was to investigate arginase release in HUS patients and laboratory models and correlate arginase levels to hemolysis and kidney injury.MethodsTwo separate cohorts of patients (n = 47 in total) with HUS associated with Shiga toxin-producing enterohemorrhagic E. coli (EHEC) and pediatric controls (n = 35) were investigated. Two mouse models were used, in which mice were either challenged intragastrically with E. coli O157:H7 or injected intraperitoneally with Shiga toxin 2. An in vitro model of thrombotic microangiopathy was developed in which Shiga toxin 2- and E. coli O157 lipopolysaccharide-stimulated human blood cells combined with ADAMTS13-deficient plasma were perfused over glomerular endothelial cells. Two group statistical comparisons were performed using the Mann–Whitney test, multiple groups were compared using the Kruskal–Wallis test followed by Dunn’s procedure, the Wilcoxon signed rank test was used for paired data, or linear regression for continuous variables.ResultsHUS patients had excessively high plasma arginase 1 levels and activity (conversion of L-arginine to urea and L-ornithine) during the acute phase, compared to remission and controls. Arginase 1 levels correlated with lactate dehydrogenase activity, indicating hemolysis, as well as the need for dialysis treatment. Patients also exhibited high levels of plasma alpha-1-microglobulin, a heme scavenger. Both mouse models exhibited significantly elevated plasma arginase 1 levels and activity. Plasma arginase 1 levels correlated with lactate dehydrogenase activity, alpha-1-microglobulin and urea levels, the latter indicative of kidney dysfunction. In the in vitro model of thrombotic microangiopathy, bioactive arginase 1 was released and levels correlated to the degree of hemolysis.ConclusionsElevated red blood cell-derived arginase was demonstrated in HUS patients and in relevant in vivo and in vitro models. The excessively high arginase levels correlated to the degree of hemolysis and kidney dysfunction. Thus, arginase inhibition should be investigated in HUS.

Investigating the presence and antibiotic susceptibilities of Escherichia coli O157 and Listeria monocytogenes in ruminant feces and feed in Balıkesir province

The aim of this study is to determine the presence, virulence factors, and antibiotic susceptibilities of E. coli O157 and L. monocytogenes in ruminant feces and feed. This study was carried out for the first time in Balıkesir province. Feces, pellet feed, and silage samples were analyzed simultaneously for E. coli O157 and L. monocytogenes using feces of one gram and feed of twenty-five grams according to ISO 16654:2001/Amd 1:2017 and ISO 11290-1, respectively. 38 (38%) E. coli O157 strains were isolated and identified from a total of 100 ruminant feces. A total of 3 (3%) E. coli O157:H7 strains were detected by PCR from one hundred ruminant fecal samples. In the study, resistance to antibiotics increased, especially in E. coli O157 isolates. In this study, enterohaemolysin was the predominant virulence factor among the E. coli isolates, and it was thought that it was important for pathogenesis. The Sxt1 gene was higher than the Stx2 gene. A total of 24 L. monocytogenes strains were isolated from a total of 100 ruminant fecal samples and 50 silage samples. Three of these strains were isolated from silage samples taken from the farms, where L. monocytogenes was isolated from sheep feces. As a result, poor quality silage could be an important source of infection for listeriosis in Balıkesir province. Epidemiologically, poor quality silage was thought to be one of the sources of listeriosis. It was thought that ruminant feces played an important role as a reservoir in the spread and transmission of E. coli O157. The antibiotic resistance status of E. coli O157 and L. monocytogenes isolates should be monitored with epidemiological studies.

Trifunctional Nanocomposites with Colorimetric Magnetic Catalytic Activities Labels in Sandwich Immunochromatographic Detection of Escherichia coli O157:H7.

The emergence of nanoenzymes has catalyzed the robust advancement of the lateral flow immunoassay (LFIA) in recent years. Among them, multifunctional nanocomposite enzymes with core-shell architectures are considered preferable for promoting the sensing ability due to their good biocompatibility, precise control over size, and surface properties etc. Herein, we developed a dual-channel ensured lateral flow immunoassay (DFLIA) platform utilizing a magnetic, colorimetric, and catalytic multifunctional nanocomposite enzyme (Fe3O4@TCPP@Pd) [TCPP, Tetrakis (4-carboxyphenyl) porphyrin] for the ultrasensitive and highly accurate rapid detection of Escherichia coli O157:H7 (E. coli O157:H7). Fe3O4@TCPP@Pd-mAb exhibits superior performance compared to traditional AuNPs, including enhanced sensitivity and an extended linear detection range, benefiting from its high brightness signal, strong magnetic separation ability, and high peroxidase activity (Vmax = 2.32 μM S1-). Moreover, the Fe3O4@TCPP@Pd-labeled mAb probe exhibited exceptional stability and high affinity toward E. coli O157:H7 (with an affinity constant of approximately 1.723 × 109 M-1), indicating its potential for the efficient capture of the pathogen. Impressively, the developed Fe3O4@TCPP@Pd-DFLIA achieved ultrasensitive detection for E. coli O157:H7 with pre- and postcatalytic naked-eye detection sensitivities of 255 cfu/mL and 77 cfu/mL, respectively, representing an approximately 41-fold improvement over the conventional AuNP-based LFIA and also possessed good specificity and reproducibility [relative standard deviation (RSD) < 10%]. Additionally, the established DFLIA exhibited satisfactory recoveries in detecting pork and milk samples, further validating the reliability of this platform for immunoassays and demonstrating its potential for utilization in bioassays and clinical diagnostics.

Extended-Spectrum β-Lactamase and carbapenemase-producing Escherichia coli O157:H7 among diarrheic patients in Shashemene, Ethiopia.

The worldwide increase in multidrug resistance is a major threat to public health. One particular concern is the presence of Escherichia coli strains that carry Extended-Spectrum β-Lactamase (ESBL) and Carbapenemase enzymes, which can make multiple antibiotics ineffective. This complicates treatment strategies and raises the risk of illness and death. The aim of this study was to isolate E. coli O157:H7, assess its susceptibility against antimicrobial agents, and determine the presence of ESBL and Carbapenemase production in stool samples collected from diarrheic patients in Shashemene, west Arsi, Ethiopia from July to November 2022. The samples were cultured McConkey Agar and E. coli were isolated and identified by standard biochemical tests using API 20E. E. coli O157:H7 was further identified using sorbitol McConkey Agar and antisera for O157 antigen test. The antimicrobial susceptibility test was performed using the Kirby-Bauer disc diffusion method using different antibiotics. Each identified isolate was screened and tested for phenotypical ESBL and Carbapenemase production using combined disc method and modified carbapenem inactivation method, respectively. Bivariant and multivariant analyses were employed using a logistic regression model for further analysis and were interpreted based on the odds ratio and level of statistical significance at a p-value <0.05 with 95% confidence interval. E. coli O157:H7 strain was found from 9% (38/423) study participants. The majority of the participants [61.9% (262/423)] were males; and 19.1% (81/ 423) of the participants were under five children. Living in urban areas, having domestic animals, and ≥5 family size in the household were identified as statistically significant factors associated with E. coli O157:H7. Twenty-seven (71.1%) and 12 (31.6%) of the 38 E. coli O157:H7 isolates were phenotypically confirmed to be ESBL and carbapenemase producers, respectively. All isolates were resistant against Ampicillin, but sensitive to ciprofloxacin. High resistance to Ampicillin and Amoxicillin/Clavulanic acid was observed among the ESBL and carbapenemase producing isolates also. The extent of detection of multidrug resistant E. coli O157:H7 isolates against three or more classes of antimicrobial agents tested was alarmingly very high (84%). The E. coli O157:H7 isolates in this study showed a significant resistance to certain antimicrobials that were tested. The level of ESBL and Carbapenemase production among these isolates was found to be quite high. We observed a high resistance to Ampicillin and Amoxicillin/Clavulanic acid among the ESBL and carbapenemase producing isolates. Ciprofloxacin was found to be the most effective drug against both the ESBL producers and nonproducers.