H7 Infection Research Articles

Intestinal tissue response to Shiga toxin exposure.

These studies have cemented the need for complex cell culture models when studying host-pathogen interactions. Common animal models such as mice are resistant to E. coli O157:H7 infections and intestinal delivery of Stx2, while humans appear to be sensitive to both. It has been proposed that in humans, shiga toxin-producing E. coli-mediated intestinal damage destroys the intestinal barrier and allows basolateral access to Stx2. In mice, there is no epithelial damage; therefore, they are resistant to epithelial delivery of Stx2 while remaining sensitive to Stx2 injection. Our studies show that like mice, the human epithelial layer is quite resistant to Stx2, and it is the sensitivity of the mesenchymal cells that kills the epithelial cells. We have shown that Stx2 is transported through the intact epithelium without causing damage to the resistant epithelial layer. Understanding tissue interactions during infections is therefore critical in determining the effects of pathogens on human tissues.

Shiga toxin-producing Escherichia coli O157:H7 outbreak associated with school field trips at a farm animal exhibit-Tennessee, September-October 2023.

In October 2023, the Tennessee Department of Health identified an outbreak of Shiga toxin-producing Escherichia coli (STEC) O157:H7 infections among elementary school students who attended school field trips to the same farm animal exhibit. Our aim was to determine STEC source and prevent additional illnesses by initiating epidemiologic, laboratory and environmental investigations. We identified cases using laboratory-based surveillance and by surveying caregivers of children who attended the exhibit. Probable cases were defined as illness with abdominal cramps or diarrhoea after attendance; confirmed cases were laboratory-confirmed STEC infection in an attendee or household contact. A site visit was conducted, and event organizers were interviewed. Human stool, animal faeces and environmental samples were tested for STEC O157:H7 by real-time polymerase chain reaction (PCR), culture and whole-genome sequencing (WGS). Approximately 2300 elementary school students attended the animal exhibit during 2 days. Field trip activities included contact with different farm animal species, drinking pasteurized milk outside animal enclosures and eating lunch in a separate building onsite. We received survey responses from 399 caregivers for 443 (19%) animal exhibit attendees. We identified 9 confirmed and 55 probable cases with illness onset dates during 26 September to 12 October. Seven children aged 1-7 years were hospitalized. Four children aged 1-6 years experienced haemolytic uraemic syndrome; none died. Laboratory testing identified STEC O157:H7 by culture from eight human stool samples with 0-1 allele difference by WGS. Three environmental samples had Shiga toxin (stx 2) genes detected by PCR, but no STEC isolates were recovered by culture. This is the largest reported STEC O157:H7 outbreak associated with an animal exhibit in Tennessee. We identified opportunities for educating school staff, event organizers and families about zoonotic disease risks associated with animal contact and published prevention measures.

Bifidobacterium longum K5 Prevents Enterohaemorrhagic Escherichia coli O157:H7 Infection in Mice through the Modulation of the Gut Microbiota.

Enterohemorrhagic Escherichia coli (EHEC) serotype O157:H7 is a commonly encountered foodborne pathogen that can cause hemorrhagic enteritis and lead to hemolytic uremic syndrome (HUS) in severe cases. Bifidobacterium is a beneficial bacterium that naturally exists in the human gut and plays a vital role in maintaining a healthy balance in the gut microbiota. This study investigated the protective effects of B. longum K5 in a mouse model of EHEC O157:H7 infection. The results indicated that pretreatment with B. longum K5 mitigated the clinical symptoms of EHEC O157:H7 infection and attenuated the increase in myeloperoxidase (MPO) activity in the colon of the mice. In comparison to the model group, elevated serum D-lactic acid concentrations and diamine oxidase (DAO) levels were prevented in the K5-EHEC group of mice. The reduced mRNA expression of tight junction proteins (ZO-1, Occludin, and Claudin-1) and mucin MUC2, as well as the elevated expression of virulence factors Stx1A and Stx2A, was alleviated in the colon of both the K5-PBS and K5-EHEC groups. Additionally, the increase in the inflammatory cytokine levels of TNF-α and IL-1β was inhibited and the production of IL-4 and IL-10 was promoted in the K5-EHEC group compared with the model group. B. longum K5 significantly prevented the reduction in the abundance and diversity of mouse gut microorganisms induced by EHEC O157:H7 infection, including blocking the decrease in the relative abundance of Roseburia, Lactobacillus, and Oscillibacter. Meanwhile, the intervention with B. longum K5 promoted the production of acetic acid and butyric acid in the gut. This study provides insights into the use of B. longum K5 for developing probiotic formulations to prevent intestinal diseases caused by pathogenic bacterial infections.

Three different routes of EHEC O157:H7 infection were used to establish EHEC broiler model

In order to study the prevention and control EHEC disease measures in poultry, the infection process and development of this disease and the pathological changes of various organs were to be observed. In this study, chickens were infected with different doses of enterohemorrhagic Escherichia coli (EHEC) O157:H7 using different routes of administration to establish EHEC broiler model. A total of 195 14-day-old broilers were randomly divided into 13 groups: including control group, Enema-drip groups (1010, 1011, 1012, 1013 CFUs E. coli O157:H7), gavage groups (P.O) (1011, 1012, 1013, 1014 CFUs E. coli O157:H7), and intraperitoneal injection group (I.P.) (108, 109, 1010, 1011 CFUs E. coli O157:H7). Escherichia coli (E. coli) was given using enema-drip, gavage or intraperitoneal infection. Then the feed intake, weight changes, stool and clinical symptoms of the chicks were recorded during the experiment. 7 d after E. coli infection, blood was collected from the jugular vein and serological tests were carried out. The liver, spleen, and colon of the chicks were extracted to get the organ index, bacteria load, and their histopathological changes. After infection with E. coli, some chicks feces were green or red watery stool, sometimes accompanied by foam, and the material to weight ratio of broilers in I.P. group increased significantly (P < 0.05), the 108 CFUs group were 1.3 times as large as control group. Three modeling methods can result in abnormal serum lipid metabolism and liver function indexes (increase of AST, TBA, T-Bil and TC level; decrease of ALB, TG, and TP level). Infection of chicks with O157:H7 by all 3 methods resulted in its detection in the liver, spleen, and colon. Three modeling methods significantly decreased liver index, and inflammatory cell infiltration and hyperemia were observed in liver. The spleen index in E. coli broilers by gavage and enema-drip was significantly decreased, splenic hyperemia and periarteriolar hyalinosis were observed. The spleen was enlarged with purplish-black spheroids in I.P. group broilers, and the spleen histological changes was more serious. The colon villi of broilers in gavage and enema-drip groups were thinner, more prone to rupture, intestinal lamina propria hyperemia, and inflammatory cell infiltration. Moreover, the number of goblet cells in the mucosal epithelium increased. E. coli O157:H7 can induce liver, spleen and intestinal damage and reduce growth performance of chicks. By comparing these 3 methods, we found that chicks infected with O157:H7 by gavage had more severe liver and intestinal damage, the enema-drip method caused most serious intestinal damage, and I.P. method significantly damaged the liver and spleen of chickens.

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.

Sığır Fekal Örneklerinden Salmonella spp. ve Escherichia coli O157:H7’ye Özgü Bakteriyofaj Varlığının Araştırılması

S. Typhimurium and E. coli O157:H7 are the most important foodborne pathogens forming bacterial biofilms that contribute to their virulence, antimicrobial resistance, and surface survival, causing severe food poisoning outbreaks worldwide. Bacteriophages are antibacterial agents that are increasingly used to control foodborne pathogens, and they also play a role in the solution against the development of antibiotic resistance. In addition, bacteriophages can be found in wastewater, natural and animal wastes, and foodstuffs. Aim of this study to determine the purification and lytic effects of Salmonella spp. and E. coli specific phages circulating in our country, which can effectively combat common Salmonella spp. and E. coli infections in our country and the world by using samples taken from the cowshed. In this study, 3 S. Typhimurium and 1 E. coli O157:H5 bacteriophages were isolated, and their lytic activities were determined. As a result, it is thought that the lytic activities of S. Typhimurium and E. coli O157:H7 bacteriophages purified from Aksaray province in this study can shed light on the treatment of S. Typhimurium and E. coli O157:H7 infections and prevention studies in the food industry.

Lcn2 deficiency accelerates the infection of Escherichia coli O157:H7 by disrupting the intestinal barrier function

Bacterial infections result in intestinal inflammation and injury, which affects gut health and nutrient absorption. Lipocalin 2 (Lcn2) is a protein that reacts to microbial invasion, inflammatory responses, and tissue damage. However, it remains unclear whether Lcn2 has a protective effect against bacterial induced intestinal inflammation. Therefore, this study endeavors to investigate the involvement of Lcn2 in the intestinal inflammation of mice infected with Enterohemorrhagic Escherichia coli O157:H7 (E. coli O157:H7). Lcn2 knockout (Lcn2−/−) mice were used to evaluate the changes of inflammatory responses. Lcn2 deficiency significantly exacerbated clinical symptoms of E. coli O157:H7 infection by reducing body weight and encouraging bacterial colonization of. Compared to infected wild type mice, infected Lcn2−/− mice had significantly elevated levels of pro-inflammatory cytokines in serum and ileum, including interleukin (IL)-6, IL-1β, and tumor necrosis factor-α (TNF-α), as well as severe villi destruction in the jejunum. Furthermore, Lcn2 deficiency aggravated intestinal barrier degradation by significantly reducing the expression of tight junction proteins occludin and claudin 1, the content of myeloperoxidase (MPO) in the ileum, and the number of goblet cells in the colon. Our findings indicated that Lcn2 could alleviate inflammatory damage caused by E. coli O157:H7 infection in mice by enhancing intestinal barrier function.

Public health risks of raw milk consumption: Lessons from a case of paediatric hemolytic uremic syndrome.

Pasteurization of raw milk is mandatory before sale in Canada and has been demonstrated to reduce the risk of food-borne illness associated with milk consumption. Consumption of raw milk sparks urgent concern from a public health perspective since it has been linked to numerous outbreaks by enteric organisms, particularly Escherichia coli-related illnesses and complications in pediatric populations. The sale and distribution of raw milk is illegal in Canada, based on these significant health risks, but growing popular interest and trends in consuming raw dairy products reflect changes in consumer preferences. Although the consumption of raw milk has been an ongoing issue, this new trend is alarming and action is needed to prevent serious consequences as seen in children and other populations with reduced immunity such as the elderly and pregnant people. This commentary explores key issues identified by a local public health unit during the investigation of a recent paediatric case of hemolytic uremic syndrome related to an E. coli O157:H7 infection that occurred within the context of consumption of raw milk. The main objective of this article is to highlight that the health risks and sequelae associated with consumption of raw milk far outweigh any potential benefits, with severe consequences particularly among children. Data and health impacts, distribution, regulation, pasteurization and proposed practice recommendations are also identified and discussed.

Notes from the Field: Multistate Outbreak of Escherichia coli O157:H7 Infections Linked to a National Fast-Food Chain - United States, 2022.

Notes from the Field: Multistate Outbreak of Escherichia coli O157:H7 Infections Linked to a National Fast-Food Chain - United States, 2022.

PLGA nanoparticles containing Intimin-Flagellin fusion protein for E. coli O157:H7 nano-vaccine

Escherichia coli O157:H7 is a foodborne pathogen that can lead to severe gastrointestinal diseases in humans. Vaccination is a promising strategy for preventing E. coli O157:H7 infections, which offers socio-economic benefits and provides the possibility of stimulating both humoral and cellular immune responses at systemic and mucosal sites. In this study, we developed a needle-free vaccine candidate against E. coli O157:H7 using poly(lactic-co-glycolic acid) (PLGA) nanoparticles entrapping a chimeric Intimin-Flagellin (IF) protein. The IF protein was expressed and verified using SDS-PAGE and western blot analysis, with a yield of 1/7 mg/L and a molecular weight of approximately 70 kDa. The prepared nanoparticles showed uniformly shaped spherical particles in the 200-nm range, as confirmed by SEM and DLS analysis. Three different routes of vaccine administration were used, including intranasal, oral, and subcutaneous, and the groups vaccinated with NPs protein had a higher antibody response compared to those receiving free protein. Subcutaneous administration of IF-NPs resulted in the highest level of IgG antibody titer, while oral administration of IF-NPs produced the highest amount of IgA antibody titer. Finally, all mice in the nanoparticle- intranasal and oral administered groups challenged with 100LD50 survived, while all control mice died before day 5. Based on these findings, we conclude that the PLGA-encapsulated IF protein has the potential to serve as a promising needle-free vaccine candidate against E. coli O157:H7.

Antibacterial characteristics of allyl methyl disulfide and dimethyl trisulfide of Allium tenuissimum flower essential oil against Escherichia coli O157:H7

The aim of this study was to compare the antibacterial properties of allyl methyl disulfide and dimethyl trisulfide with the chemical compositions of Allium tenuissimum flower essential oil against Escherichia coli O157:H7. Allium tenuissimum flower essential oil was extracted from fresh Allium tenuissimum flower by hydro-distillation method. Its antibacterial activities against Escherichia coli O157:H7, Staphylococcus aureus and Listeria monocytogenes were determined by diameters of the inhibition zone. Escherichia coli O157:H7 treated with the essential oil had the largest diameter of the inhibition zone (15.2 mm) among the three strains. Allyl methyl disulfide and dimethyl trisulfide were two main compounds of the essential oil, as analyzed by gas chromatography-mass spectrometry. The antibacterial characteristics of these two compounds against Escherichia coli O157:H7 were evaluated by minimum inhibitory and bactericidal concentrations, morphologies, cytomembrane destructions, biomacromolecule contents, and metabolic enzyme activities. Minimum inhibitory concentrations of allyl methyl disulfide and dimethyl trisulfide were both 0.7 mg/mL, and minimum bactericidal concentrations of these two compounds were both 1.4 mg/mL. The images of atomic force microscopy exhibited that the surface structures and cytomembranes of Escherichia coli O157:H7 cells were rigorously damaged by 1.4 mg/mL of allyl methyl disulfide and dimethyl trisulfide, leading to the change of β-galactosidases and the leakage of cellular nucleic acids from the bacterial cells. Moreover, the intensities of deoxyribonucleic acid and protein bands of the bacterial cells were significantly decreased after treated with these two compounds at 1.4 mg/mL. Adenosine triphosphatase activities (3.75 and 1.97 U/mg prot) of the bacterial cells treated with allyl methyl disulfide (1.4 mg/mL) and dimethyl trisulfide (1.4 mg/mL) were distinctly lower than the activity of the control (6.05 U/mg prot). Allyl methyl disulfide (1.4 mg/mL) and dimethyl trisulfide (1.4 mg/mL) inactivated 84.20 % and 90.76 % respiratory-chain dehydrogenase of the bacterial cells, respectively. These results demonstrated that allyl methyl disulfide and dimethyl trisulfide exhibited efficient antibacterial characteristics against Escherichia coli O157:H7 and could be used as natural antibacterial agents in the prevention of Escherichia coli O157:H7 infection.

L-Arabinose inhibits Shiga toxin type 2-converting bacteriophage induction in Escherichia coli O157:H7

ABSTRACT The pathogenicity of Escherichia coli (E. coli) O157:H7 is predominantly associated with Shiga toxin 2 (Stx2) that poses a huge threat to human and animal intestinal health. Production of Stx2 requires expression of stx2 gene, which is located in the genome of lambdoid Stx2 prophage. Growing evidence has implicated that many commonly consumed foods participate in the regulation of prophage induction. In this study, we aimed to explore whether specific dietary functional sugars could inhibit Stx2 prophage induction in E. coli O157:H7, thereby preventing Stx2 production and promoting intestinal health. We demonstrated that Stx2 prophage induction in E. coli O157:H7 was strongly inhibited by L-arabinose both in vitro and in a mouse model. Mechanistically, L-arabinose at doses of 9, 12, or 15 mM diminished RecA protein levels, a master mediator of the SOS response, contributing to reduced Stx2-converting phage induction. L-Arabinose inhibited quorum sensing and oxidative stress response, which are known as positive regulators of the SOS response and subsequent Stx2 phage production. Furthermore, L-arabinose impaired E. coli O157:H7 arginine transport and metabolism that were involved in producing Stx2 phage. Collectively, our results suggest that L-arabinose may be exploited as a novel Stx2 prophage induction inhibitor against E. coli O157:H7 infection.

Enterohemorrhagic Escherichia coli senses microbiota-derived nicotinamide to increase its virulence and colonization in the large intestine

Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is a foodborne pathogen that specifically colonizes and infects the human large intestine. EHEC O157:H7 engages intricate regulatory pathways to detect host intestinal signals and regulate virulence-related gene expression during colonization and infection. However, the overall EHEC O157:H7 virulence regulatory network in the human large intestine remains incompletely understood. Here, we report a complete signal regulatory pathway where the EvgSA two-component system responds to high-nicotinamide levels produced by microbiota in the large intestine and directly activates loci of enterocyte effacement genes to promote EHEC O157:H7 adherence and colonization. This EvgSA-mediated nicotinamide signaling regulatory pathway is conserved and widespread among several other EHEC serotypes. Moreover, disruption of this virulence-regulating pathway by the deletion of evgS or evgA significantly decreased EHEC O157:H7 adherence and colonization in the mouse intestinal tract, indicating that these genes could be potential targets for the development of new therapeutics for EHEC O157:H7 infection.

Antibody prevalence to avian influenza virus subtypes H5, H7 and H9 in falcons, captive and wild birds, United Arab Emirates, 2003-2006.

Avian influenza viruses (AIV) may cause enormous economic losses in the poultry industry and sporadically severe disease in humans. Falconry is a tradition of great importance in the Arabian Peninsula. Falcons may catch AIV through contact with infected quarry species. Falcons together with other bird species are the focus of this seroprevalence study, carried out on sera collected in the United Arab Emirates (UAE). AIV with the haemagglutinin subtypes H5, H7 and possibly H9 may infect humans. We investigated the antibody prevalence to these subtypes in falcons and other birds by haemagglutination inhibition test. 617 sera of falcons and 429 sera of 46 wild/captive bird species were tested. From the falcons, only one was positive for H5 antibodies (0.2%), none contained antibodies to H7, but 78 had antibodies to H9 (13.2%). Regarding other birds, eight were positive for antibodies to H5 (2.1%), none had antibodies to H7, but 55 sera from 17 species contained antibodies to H9 (14.4%). In contrast to H5 and H7 infections, H9N2 is widespread worldwide. Its ability to reassort, thereby creating possibly pathogenic strains for humans, should remind us of the potential risk that close contact with birds entails.

Impact of Selenium Nanoparticle-Enriched Lactobacilli Feeding Against Escherichia coli O157:H7 Infection of BALB/c Mice.

The effectiveness of selenium nanoparticle (SeNP)-enriched Lactiplantibacillus plantarum and Lactobacillus acidophilus was studied against Shiga toxin-producing Escherichia coli O157:H7 infection on the intestinal fragments and kidney tissue of BALB/c mice. Gut microbiota-targeted bacteria and E. coli O157:H7 counts were obtained by qPCR and PCR. Histology of ileum, colon, and kidney tissues and Stx secretions were analyzed until one-week post-infection. Mice fed with SeNP Lpb. plantarum in the preinfection feeding groups have lower E. coli O157:H7 counts and lower intestinal damage than those in the infected group. The lowest mean fecal probiotic counts were in the L. acidophilus group (7.61 log 10). In pretreatment groups of SeNP L. acidophilus and L. acidophilus, the mean counts of bacteria decreased to 104CFU/g by day 7. The lowest Stx copy number was demonstrated in SeNP Lpb. plantarum feeding groups' day 7 (P < 0.05). Feeding groups with SeNP Lpb. plantarum had significantly higher members of Lactobacilli in their fecal microbiota than the control group on day 7. It was clarified that Se-enriched Lpb. plantarum and L. acidophilus can be useful as a method of preventing STEC infections. The viability of STEC infection exposure to selenium-enriched Lactobacillus spp. was decreased more than for non-Se-enriched Lactobacillus spp.

Cross contamination of Escherichia coli O157:H7 in fresh-cut leafy vegetables: Derivation of a food safety objective and other risk management metrics

In the present study, the distribution of Escherichia coli O157:H7 was investigated in fresh-cut lettuce by simulating a commercial process at a pilot plant scale with different initial inoculum levels. A deterministic approach was used to derive a potential Food Safety Objective (FSO) for the studied pathogen. The experimental outcomes, together with literature data, were used to develop a probabilistic exposure model for E. coli O157:H7 to elucidate potential risk management metrics, i.e., Performance Objectives (POs) and a Performance Criterion (PC) at specific process steps within the food chain of fresh-cut leafy vegetables. The proposed FSO was estimated to be −6.0 log CFU/g for E. coli O157:H7 at consumption. The experimental results indicated that the pathogen was homogeneously distributed in fresh-cut lettuce after processing. Based on the modelling results, the average concentration estimated as PO in unprocessed lettuce should be −5.8 log CFU/g to meet the proposed FSO. On the other hand, the PO established after washing and packaging (processed lettuce) showed a lower value, with −6.8 log CFU/g E. coli O157:H7. Reduction levels during the washing or disinfection step estimated as the PC should be > 2.9 log CFU/g. The establishment of PC for the washing step could be considered as a suitable measure to reach the FSO in fresh-cut leafy vegetables. The suggested POs and PC would help food businesses and authorities to establish safety targets and corrective actions to reduce the number of potentially contaminated lettuce lots on the market, thus reducing the number of E. coli O157:H7 infections.

P22-Based Nanovaccines against Enterohemorrhagic Escherichia coli.

Enterohemorrhagic Escherichia coli (EHEC) is an important causative agent of diarrhea in humans that causes outbreaks worldwide. Efforts have been made to mitigate the morbidity and mortality caused by these microorganisms; however, the global incidence is still high, causing hundreds of deaths per year. Several vaccine candidates have been evaluated that demonstrate some stability and therapeutic potential but have limited overarching effect. Virus-like particles have been used successfully as nanocontainers for the targeted delivery of drugs, proteins, or nucleic acids. In this study, phage P22 nanocontainers were used as a carrier for the highly antigenic T3SS structural protein EscC that is conserved between EHEC and other enteropathogenic bacteria. We were able to stably incorporate the EscC protein into P22 nanocontainers. The EscC-P22 particles were used to intranasally inoculate mice, which generated specific antibodies against EscC. These antibodies increased the phagocytic activity of murine macrophages infected with EHEC in vitro and reduced bacterial adherence to Caco-2 epithelial cells in vitro, illustrating their functionality. The EscC-P22-based particles are a potential nanovaccine candidate for immunization against EHEC O157:H7 infections. IMPORTANCE This study describes the initial attempt to use P22 viral-like particles as nanocontainers expressing enterohemorrhagic Escherichia coli (EHEC) proteins that are immunogenic and could be used as effective vaccines against EHEC infections.

Virtual Screening and In Vitro Experimental Verification of LuxS Inhibitors for Escherichia coli O157:H7.

Enterohemorrhagic Escherichia coli O157:H7 is an important foodborne pathogen that forms biofilms. In this study, three quorum-sensing (QS) inhibitors (M414-3326, 3254-3286, and L413-0180) were obtained through virtual screening, and their in vitro antibiofilm activities were validated. Briefly, the three-dimensional structure model of LuxS was constructed and characterized using the SWISS-MODEL. High-affinity inhibitors were screened from the ChemDiv database (1,535,478 compounds) using LuxS as a ligand. Five compounds (L449-1159, L368-0079, M414-3326, 3254-3286, and L413-0180) with a good inhibitory effect (50% inhibitory concentration <10 μM) on type II QS signal molecule autoinducer-2 (AI-2) were obtained using a AI-2 bioluminescence assay. The absorption, distribution, metabolism, excretion, and toxicity (ADMET) properties predicated that the five compounds had high intestinal absorption levels (high) and plasma protein binding (absorbent strong) and did not inhibit the metabolism of CYP2D6 metabolic enzymes. In addition, molecular dynamics simulation showed that compounds L449-1159 and L368-0079 could not stably bind with LuxS. Thus, these compounds were excluded. Furthermore, surface plasmon resonance results showed that the three compounds could specifically bind to LuxS. IN addition, the three compounds could effectively inhibit the biofilm formation without affecting the growth and metabolism of the bacteria. Finally, the reverse transcription-quantitative PCR results showed that the three compounds downregulated the expression of the LuxS gene. Overall, these results revealed that the three compounds obtained through virtual screening could inhibit biofilm formation of E. coli O157:H7 and are potential LuxS inhibitors that can be used to treat E. coli O157:H7 infections. IMPORTANCE E. coli O157:H7 is a foodborne pathogen of public health importance. Quorum sensing (QS) is a form of bacterial communication that can regulate various group behaviors, including biofilm formation. Here, we identified three QS AI-2 inhibitors (M414-3326, 3254-3286, and L413-0180) that can stably and specifically bind to LuxS protein. The three QS AI-2 inhibitors inhibited biofilm formation without affecting the growth and metabolic activity of E. coli O157:H7. The three QS AI-2 inhibitors are promising agents for treating E. coli O157:H7 infections. Further studies to identify the mechanism of the three QS AI-2 inhibitors are needed to develop new drugs to overcome antibiotic resistance.

Visual and Rapid Detection of Escherichia coli O157:H7 in Stool Samples by FTA Card-based Loop-mediated Isothermal Amplification

Objective: Escherichia coli O157:H7 (E. coli O157:H7) can induce severe diseases in animals and humans that result in significant public health problems. Therefore, the development of rapid and visual detection methods to diagnose E. coli O157:H7 infections and monitor its prevalence is critical for the prevention and control purposes. Methods: A colorimetric loop-mediated isothermal amplification (LAMP) assay was utilized to detect E. coli O157:H7. A DNA extraction kit and Flinders Technology Associates (FTA) cards were used to extract nucleic acid in conjunction with colorimetric LAMP detection. Results: The method developed effectively distinguished E. coli O157:H7 from other pathogens with a detection limit of 25 CFU/mL in spiked stool samples. In addition, the nucleic acid of these samples was easily extracted and transported with an FTA card at room temperature. The entire detection process was completed within 35 min using simple constant-temperature equipment. Conclusion: The colorimetric LAMP method with FTA card-based nucleic acid purification was shown to rapidly detect E. coli O157:H7 with sensitivity and specificity. This visual method is expected to be widely used to control E. coli O157:H7 infections, particularly in resource-limited settings.

Quantitative risk assessment-epidemic curve prediction model for leafy green outbreak investigation.

The objective of this study was to leverage quantitative risk assessment to investigate possible root cause(s) of foodborne illness outbreaks related to Shiga toxin-producing Escherichia coli O157:H7 (STEC O157) infections in leafy greens in the United States. To this end, we developed the FDA leafy green quantitative risk assessment epidemic curve prediction model (FDA-LG QRA-EC) that simulated the lettuce supply chain. The model was used to predict the number of reported illnesses and the epidemic curve associated with lettuce contaminated with STEC O157 for a wide range of scenarios representing various contamination conditions and facility processing/sanitation practices. Model predictions were generated for fresh-cut and whole lettuce, quantifying the differing impacts of facility processing and home preparation on predicted illnesses. Our model revealed that the timespan (i.e., number of days with at least one reported illness) and the peak (i.e., day with the most predicted number of reported illnesses) of the epidemic curve of a STEC O157-lettuce outbreak were not strongly influenced by facility processing/sanitation practices and were indications of contamination pattern among incoming lettuce batches received by the facility or distribution center. Through comparisons with observed number of illnesses from recent STEC O157-lettuce outbreaks, the model identified contamination conditions on incoming lettuce heads that could result in an outbreak of similar size, which can be used to narrow down potential root cause hypotheses.