Investigation of Feedlot-level Use of a Direct-fed Microbial on Fecal Shedding of E. coli O157:H7.

Our objectives were to determine whether the feedlot-level use of a direct-fed microbial (DFM; Lactobacillus animalis LA51 and Propionibacterium freudenreichii PF24; Bovamine Defend®, 2 × 109 CFU/g) was associated with fecal prevalence and concentration of E. coli O157:H7, and determine pen- and feedlot-level risk factors associated with fecal E. coli O157:H7 prevalence in cattle pens from commercial feedlot operations. Twenty commercial feedlots in Nebraska, ten that included DFM (DFM) and ten that did not (no-DFM), were sampled during the summer of 2017. In each sampling month, 22 pen-floor fecal samples were collected from three pens in each feedlot. Samples were subjected to cultural and molecular procedures for the detection of E. coli O157:H7 (immunomagnetic separation, plating on selective media, followed by PCR confirmation) and spiral plating for quantification. A total of 1,320 samples from 180 pens of finishing cattle belonging to 20 feedlots, which were sampled three times throughout a 12-week period, were processed and tested. Across all feedlots and sampling months, the mean within-pen prevalence was 13.5% (95% CI = 2.6-47.4%). The association between DFM status and the within-pen prevalence of E. coli O157:H7 depended significantly (p < 0.05) on the sampling month. The second sampling month between late July and mid-August corresponded to the highest within-pen prevalence estimates reported in this study, with no-DFM pens having a higher prevalence than DFM pens. After accounting for the DFM status, and based on multivariable analyses, sampling month, average pen body weight, and weather conditions were significantly associated with the within-pen fecal prevalence of E. coli O157:H7. Collectively, these findings demonstrate that the use of a DFM containing Lactobacillus animalis LA51 and Propionibacterium freudenreichii PF26 in feedlots showed potential in reducing fecal E. coli O157:H7 prevalence in cattle during times when prevalence peaks.

Models for the Inactivation of Foodborne Pathogens in Salad Dressing from Challenge Studies.

The Association for Dressings and Sauces' (ADS) members have conducted challenge studies on salad dressing products to assess pathogen survival. Data from 79 different challenge studies provided by ADS were used in this analysis. The acid-moisture ratio, pH, incubation temperature and ingredient details were provided for each study. Linear regression models were used to predict the time to 3-log, 4-log, and 5-log reduction as a function of study parameters. A statistically based approach also was used to estimate the concentration of pathogens in ingredients based on testing history. This was combined with decline modeling to estimate pathogen concentration over time. The time to five log reduction for each of the target pathogens were highly skewed. A logarithmic transformation of time to 5 log reduction resulted in approximately normally distributions. Incubation temperature and formulation pH were highly significant (p < 1E-6), in predicting the number of days to a five-log reduction of Escherichia coli O157:H7, while the percentage of spices in the formulation is also quite significant (p = 0.01). Salmonella modeling showed that the most highly significant parameter was the percentage of water (p < 1E-8). Other parameters in order of descending significance include the percent fruit (p=0.00032), incubation temperature (p=0.00268), followed by percent sugar (p=0.02161) and percent vegetables (p=0.03149). The most significant parameter in predicting Listeria monocytogenes reduction was incubation temperature (p=0.000687), followed by acid moisture ratio (p=0.012423). The next two significant parameters in the Listeria model were percent lipid (p=0.023772) and percent water (p=0.025701). The least significant parameter that meets the minimum criteria for inclusion in the Listeria model (p<0.05) was percent fruit (p=0.047074). Our analysis will be useful in developing risk-based approaches to continue to assure the safety of commercially prepared salad dressings.

Antimicrobial Resistance in Diverse Escherichia coli Pathotypes from Nigeria.

Escherichia coli is a gram-negative commensal bacterium living in human and animal intestines. Its pathogenic strains lead to high morbidity and mortality, which can adversely affect people by causing urinary tract infections, food poisoning, septic shock, or meningitis. Humans can contract E. coli by eating contaminated food-such as raw or undercooked raw milk, meat products, and fresh produce sold in open markets-as well as by coming into contact with contaminated settings like wastewater, municipal water, soil, and faeces. Some pathogenic strains identified in Nigeria, include Enterohemorrhagic (Verotoxigenic), Enterotoxigenic, Enteropathogenic, Enteroinvasive, and Enteroaggregative E. coli. This causes acute watery or bloody diarrhoea, stomach cramps, and vomiting. Apart from the virulence profile of E. coli, antibiotic resistance mechanisms such as the presence of blaCTX-M found in humans, animals, and environmental isolates are of great importance and require surveillance and monitoring for emerging threats in resource-limited countries. This review is aimed at understanding the underlying mechanisms of evolution and antibiotic resistance in E. coli in Nigeria and highlights the use of improving One Health approaches to combat the problem of emerging infectious diseases.

Lipopolysaccharide Core Truncation in Invasive Escherichia coli O157:H7 ATCC 43895 Impairs Flagella and Curli Biosynthesis and Reduces Cell Invasion Ability.

Escherichia coli O157:H7 (E. coli O157) is known for causing severe foodborne illnesses such as hemorrhagic colitis and hemolytic uremic syndrome. Although E. coli O157 is typically regarded as an extracellular pathogen and a weak biofilm producer, some E. coli O157 strains, including a clinical strain ATCC 43895, exhibit a notable ability to invade bovine crypt cells and other epithelial cells, as well as to form robust biofilm. This invasive strain persists in the bovine host significantly longer than non-invasive strains. Various surface-associated factors, including lipopolysaccharides (LPS), flagella, and other adhesins, likely contribute to this enhanced invasiveness and biofilm formation. In this study, we constructed a series of LPS-core deletion mutations (waaI, waaG, waaF, and waaC) in E. coli O157 ATCC 43895, resulting in stepwise truncations of the LPS. This approach enabled us to investigate the effects on the biosynthesis of key surface factors, such as flagella and curli, and the ability of this invasive strain to invade host cells. We confirmed the LPS structure and found that all LPS-core mutants failed to form biofilms, highlighting the crucial role of core oligosaccharides in biofilm formation. Additionally, the LPS inner-core mutants ΔwaaF and ΔwaaC lost the ability to produce flagella and curli. Furthermore, these inner-core mutants exhibited a dramatic reduction in adherence to and invasion of epithelial cells (MAC-T), showing an approximately 100-fold decrease in cell invasion compared with the outer-core mutants (waaI and waaG) and the wild type. These findings underscore the critical role of LPS-core truncation in impairing flagella and curli biosynthesis, thereby reducing the invasion capability of E. coli O157 ATCC 43895.

Isolation and Characterization of Novel Escherichia coli O157:H7 Phage SPEC13 as a Therapeutic Agent for E. coli Infections In Vitro and In Vivo.

Escherichia coli O157:H7 is a recognized food-borne pathogen causing severe food poisoning at low doses. Bacteriophages (phages) are FDA-approved for use in food and are suggested as natural preservatives against specific pathogens. A novel phage must be identified and studied to develop a new natural preservative or antimicrobial agent against E. coli O157:H7. The phage SPEC13 displayed broad host range and was classified within the Ackermannviridae family based on its observed characteristics by a TEM and genome analysis. In 10 min, this phage achieves a remarkable 93% adsorption rate with the host. Its latency period then lasts about 20 min, after which it bursts, releasing an average of 139 ± 3 PFU/cell. It exhibited robustness within a pH range of 4 to 12, indicating resilience under diverse environmental circumstances. Furthermore, SPEC13 demonstrated stability at an ambient temperature up to 60 °C. A whole genome and phylogenetics analysis revealed that SPEC13 is a novel identified phage, lacking a lysogenic life cycle, antibiotic resistance genes, or genes associated with virulence, thereby presenting a promising biological agent for therapeutic application. Animal studies showed that SPEC13 effectively controlled the growth of harmful bacteria, resulting in a significant improvement in colon health, marked by reduced swelling (edema) and tissue damage (mucosal injury). The introduction of SPEC13 resulted in a substantial decrease in quantities of E. coli O157:H7, reducing the bacterial load to approximately 5 log CFU/g of feces. In conclusion, SPEC13 emerges as a promising inclusion in the array of phage therapy, offering a targeted and efficient approach for addressing bacterial infections.

Portable real-time determination of Escherichia coli O157:H7 and Staphylococcus aureus based on smartphones and hydrogels.

The aptamers functionalized orange-emission carbon dots (OCDs) and green-emission carbon dots (GCDs) had dual-emission peaks with single excitation. Tungsten disulfide nanosheets (WS2 NSs)-triggered fluorescence quenching achieved the ratiometric fluorescence determination of Escherichia coli O157:H7 (E. coli O157:H7) and Staphylococcus aureus (S. aureus) with wide ranges of 18-1.8 × 106 and 37-3.7 × 107 CFU/mL and low detection limits of 8 and 20 CFU/mL, respectively. The results in real sample with recoveries of 90-101 % and RSD < 4.12 % were no significant difference from standard plate counting method. Meanwhile, the dual-color CDs were further adopted in the smartphone-assisted hydrogel platform and achieved speedy, sensitive, portable and real-time determination of E. coli O157:H7 and S. aureus in real samples. This work has not only developed ratiometric fluorescence detection and constructed a portable hydrogel platform, but also provided a unique strategy in developing a time-efficient and easy-to-use portable device in food safety monitoring.

Survival of Twelve Pathogenic and Generic Escherichia coli Strains in Agricultural Soils as Influenced by Strain, Soil Type, Irrigation Regimen, and Soil Amendment.

Biological soil amendments of animal origin (BSAAO) play an important role in agriculture but can introduce pathogens into soils. Pathogen survival in soil is widely studied, but data are needed on the impacts of strain variability and field management practices. This study monitored the population of 12 Escherichia coli strains (generic, O157, and non-O157) in soils while evaluating the interactions of soil type, irrigation regimen, and soil amendment in three independent, greenhouse-based, randomized complete block design trials. Each E. coli strain (4-5 log10 CFU/g) was homogenized in bovine manure amended or nonamended sandy-loam or clay-loam soil. E. coli was enumerated in 25 g samples on 0, 0.167 (4 h), 1, 2, 4, 7, 10, 14, 21, 28, 56, 84, 112, 168, 210, 252, and 336 days postinoculation (dpi). Regression analyses were developed to understand the impact of strain, soil type, irrigation regimen, and soil amendment on inactivation rates. E. coli survived for 112 to 336 dpi depending on the treatment combination. Pathogenic and generic E. coli survived 46 days [95% Confidence interval (CI) = 20.85, 64.72; p = 0.001] longer in soils irrigated weekly compared to daily and 146 days (CI = 114.50, 184.50; p < 0.001) longer in amended soils compared to unamended soils. Pathogenic E. coli strains were nondetectable 69 days (CI = 39.58, 98.66, p = 0.015) earlier than generic E. coli strains. E. coli inactivation rates demonstrated a tri-phasic pattern, with breakpoints at 26 dpi (CI = 22.3, 29.2) and 130 dpi (CI = 121.0, 138.1). The study findings demonstrate that using bovine manure as BSAAO in soil enhances E. coli survival, regardless of strain, and adequate food safety practices are needed to reduce the risk of crop contamination. The findings of this study contribute data on E. coli concentrations in amended soils to assist stakeholders and regulators in making risk-based decisions on time intervals between the application of BSAAO and the production and harvest of fruits and vegetables.

Impact of Operational Parameters on Pathogen Lethality in Dry and Semi-dry Uncooked Fermented Sausages.

The safety of uncooked fermented, dried sausages relies upon controlled fermentation and drying that inactivates pathogenic bacteria. Current guidelines for the production of fermented sausages by the United States Department of Agriculture (USDA) Food Safety Inspection Services (FSIS) and related research highlight specific safety parameters. The confidence that processing steps, which do not include cooking, inherently mitigate microbial risks, is challenged by the resilience of pathogens in the dry and acidic environments of these food products. The aim of this work was to examine the length of drying required to achieve a target pathogen reduction across a range of sausage diameters. This study investigated the relationship between product diameter and time required to achieve target reductions of Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes, as well as the attainment of specific water activity (aw). The research utilized salami and summer sausage with diameters of 18 mm, 30 mm, 60 mm, 90 mm, and 110 mm. Sausage batter was inoculated with 5 strains each of E. coli O157:H7, L. monocytogenes, and S. enterica. Inoculated sausages were processed with fermentation and drying protocols for each sausage type. Smaller diameter sausages reached both the desired pathogen reduction and target aw of 0.85 sooner than larger ones. However, the time to achieve the target aw did not align with the time to achieve the pathogen reduction targets, suggesting that aw alone is not a reliable indicator of safety. Another finding was larger sausages achieved the target pathogen reduction without reaching the target aw, suggesting complex relationship between aw, diameter, and pathogen inactivation. These data support the need for food safety guidelines that consider drying duration, aw, and pathogen behavior for varying sausage diameters. This research contributes to developing more precise safety protocols for producing dry and semi-dry fermented sausages.

A ratiometric SERS aptasensor based on catalytic hairpin self-assembly mediated cyclic signal amplification strategy for the reliable determination of E. coli O157:H7.

A ratiometric SERS aptasensor based on catalytic hairpin self-assembly (CHA) mediated cyclic signal amplification strategy was developed for the rapid and reliable determination of Escherichia coli O157:H7. The recognition probe was synthesized by modifying magnetic beads with blocked aptamers, and the SERS probe was constructed by functionalizing gold nanoparticles (Au NPs) with hairpin structured DNA and 4-mercaptobenzonitrile (4-MBN). The recognition probe captured E. coli O157:H7 specifically and released the blocker DNA, which activated the CHA reaction on the SERS probe and turned on the SERS signal of 6-carboxyl-x-rhodamine (ROX). Meanwhile, 4-MBN was used as an internal reference to calibrate the matrix interference. Thus, sensitive and reliable determination and quantification of E. coli O157:H7 was established using the ratio of the SERS signal intensities of ROX to 4-MBN. This aptasensor enabled detection of 2.44 × 102 CFU/mL of E. coli O157:H7 in approximately 3 h without pre-culture and DNA extraction. In addition, good reliability and excellent reproducibility were observed for the determination of E. coli O157:H7 in spiked water and milk samples. This study offered a new solution for the design of rapid, sensitive, and reliable SERS aptasensors.

Hybrid nanoflower-based electrochemical lateral flow immunoassay for Escherichia coli O157 detection.

An electrochemical biosensor has been developed for detection of Escherichia coli O157 by integrating lateral flow with screen-printed electrodes. The screen-printed electrodes were attached under the lateral flow detection line, and organic-inorganic nanoflowers prepared from E. coli O157-specific antibodies as an organic component were attached to the lateral flow detection line. In the presence of E. coli O157, an organic-inorganic nanoflower-E. coli O157-antimicrobial peptide-labelled ferrocene sandwich structure is formed on the lateral flow detection line. Differential pulse voltammetry is applied using a smartphone-based device to monitor ferrocene on the detection line. The resulting electrochemical biosensor could specifically detect E. coli O157 with a limit of detection of 25 colony-forming units mL-1. Through substitution of antibodies of organic components in organic-inorganic nanoflowers, biosensors have great potential for the detection of other pathogens in biomedical research and clinical diagnosis.

Vibrio cholerae O1 and Escherichia coli O157:H7 from drinking water and wastewater in Addis Ababa, Ethiopia.

BACKGROUND: In Addis Ababa, Ethiopia, open ditches along innner roads in residential areas serve to convey domestic wastewater and rainwater away from residences. Contamination of drinking water by wastewater through faulty distribution lines could expose households to waterborne illnesses. This prompted the study to assess the microbiological safety of wastewater and drinking water in Addis Ababa, identify the pathogens therein, and determine their antibiotic resistance patterns. RESULTS VIBRIO CHOLERAE: O1, mainly Hikojima serotype, was isolated from 23 wastewater and 16 drinking water samples. Similarly, 19 wastewater and 10 drinking water samples yielded Escherichia coli O157:H7. V. cholerae O1 were 100% resistant to the penicillins (Amoxacillin and Ampicillin), and 51-82% were resistant to the cephalosporins. About 44% of the V. cholerae O1 isolates in this study were Extended Spectrum Beta-Lactamase (ESBL) producers. Moreover, 26% were resistant to Meropenem. Peperacillin/Tazobactam was the only effective ß-lactam antibiotic against V. cholerae O1. V. cholerae O1 isolates showed 37 different patterns of multiple resistance ranging from a minimum of three to a maximum of ten antimicrobials. Of the E. coli O157:H7 isolates, 71% were ESBL producers. About 96% were resistant to Ampicillin. Amikacin and Gentamicin were very effective against E. coli O157:H7 isolates. The isolates from wastewater and drinking water showed multiple antibiotic resistance against three to eight antibiotic drugs. CONCLUSIONS: Open ditches for wastewater conveyance along innner roads in residence areas and underground faulty municipal water distribution lines could be possible sources for V. cholerae O1 and E. coli O157:H7 infections to surrounding households and for dissemination of multiple drug resistance in humans and, potentially, the environment.

Quantitative microbial risk assessment for Escherichia Coli O157: H7 via drinking water in the Gaza Strip, Palestine.

Introduction: Microbial contamination of drinking water, particularly by pathogens such as Escherichia coli O157: H7, is a significant public health concern worldwide, especially in regions with limited access to clean water like the Gaza Strip. However, few studies have quantified the disease burden associated with E. coli O157: H7 contamination in such challenging water management contexts. Objective: This study aimed to conduct a comprehensive Quantitative Microbial Risk Assessment to estimate the annual infection risk and disease burden attributed to E. coli O157: H7 in Gaza's drinking water. Methods: Applying the typical four steps of the Quantitative Microbial Risk Assessment technique-hazard identification, exposure assessment, dose-response analysis, and risk characterization-the study assessed the microbial risk associated with E. coli O157: H7 contamination in Gaza's drinking water supply. A total of 1317 water samples from various sources across Gaza were collected and analyzed for the presence of E. coli O157: H7. Using Microsoft ExcelTM and @RISKTM software, a Quantitative Microbial Risk Assessment model was constructed to quantify the risk of infection associated with E. coli O157: H7 contamination. Monte Carlo simulation techniques were employed to assess uncertainty surrounding input variables and generate probabilistic estimates of infection risk and disease burden. Results: Analysis of the water samples revealed the presence of E. coli O157: H7 in 6.9% of samples, with mean, standard deviation, and maximum values of 1.97, 9.74, and 112 MPN/100 ml, respectively. The risk model estimated a median infection risk of 3.21 × 10-01 per person per year and a median disease burden of 3.21 × 10-01 Disability-Adjusted Life Years per person per year, significantly exceeding acceptable thresholds set by the WHO. Conclusion: These findings emphasize the urgent need for proactive strategies to mitigate public health risks associated with waterborne pathogens in Gaza.

Structure of plasmonic multi spectral Apta sensor and analyzing of bulk and surface sensitivity.

In this work, a multispectral aptasensor structure, including a sub-layer and two side walls, was presented. The cells are positioned at the down and top of the structure, with the down cells oriented perpendicular to the walls and the top cells aligned parallel to the walls. The validity of the findings was verified by the utilization of a numerical simulation technique known as 3D Finite Difference Time Domain (FDTD). The biosensor under consideration exhibits sensitivities of 1093.7 nm/RIU, 754 nm/RIU, and 707.43 nm/RIU in mode III, mode II, and mode I, respectively. In the majority of instances, the quantity of analyte available is insufficient to coat the surface of the sensor thoroughly. Consequently, in this study, the evaluation of surface sensitivity was undertaken alongside bulk sensitivity. The surface sensitivity of the suggested structure for mode II in the sensor layer, with thicknesses of 10, 20, 30, and 70 nm, is measured to be 25, 78, 344, and 717.636 nm/RIU, respectively. Our design incorporates a unique arrangement of sub-layer and side walls, with cells positioned to maximize interaction with the target analyte. This innovative configuration, combined with Ag for its superior plasmonic properties, enables the detection of E. coli O157 with remarkable sensitivity.

Natural approach of using nisin and its nanoform as food bio-preservatives against methicillin resistant Staphylococcus aureus and E.coli O157:H7 in yoghurt.

BACKGROUND: Natural antimicrobial agents such as nisin were used to control the growth of foodborne pathogens in dairy products. The current study aimed to examine the inhibitory effect of pure nisin and nisin nanoparticles (nisin NPs) against methicillin resistant Staphylococcus aureus (MRSA) and E.coli O157:H7 during the manufacturing and storage of yoghurt. Nisin NPs were prepared using new, natural, and safe nano-precipitation method by acetic acid. The prepared NPs were characterized using zeta-sizer and transmission electron microscopy (TEM). In addition, the cytotoxicity of nisin NPs on vero cells was assessed using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The minimum inhibitory concentrations (MICs) of nisin and its nanoparticles were determined using agar well-diffusion method. Further, fresh buffalo's milk was inoculated with MRSA or E.coli O157:H7 (1 × 106 CFU/ml) with the addition of either nisin or nisin NPs, and then the inoculated milk was used for yoghurt making. The organoleptic properties, pH and bacterial load of the obtained yoghurt were evaluated during storage in comparison to control group. RESULTS: The obtained results showed a strong antibacterial activity of nisin NPs (0.125 mg/mL) against MRSA and E.coli O157:H7 in comparison with control and pure nisin groups. Notably, complete eradication of MRSA and E.coli O157:H7 was observed in yoghurt formulated with nisin NPs after 24 h and 5th day of storage, respectively. The shelf life of yoghurt inoculated with nisin nanoparticles was extended than those manufactured without addition of such nanoparticles. CONCLUSIONS: Overall, the present study indicated that the addition of nisin NPs during processing of yoghurt could be a useful tool for food preservation against MRSA and E.coli O157:H7 in dairy industry.

Fluorescent biosensor based on magnetic separation platform and spore-like breakable organosilica nanocapsules controlled-release carbon dots for the detection of Escherichia coli O157:H7.

Ultrasensitive and rapid detection of low concentration of Escherichia coli O157: H7 (E. coli O157:H7) in food is essential for food safety and public health. In this study, A novel fluorescence signal amplification biosensor based on magnetic separation platform and red fluorescent carbon dots (R-CDs)-encapsulated breakable organosilica nanocapsules (BONs) for ultrasensitive detection of E. coli O157:H7 was established. Wulff-type boronic acid functionalized magnetic nanoparticles (MNPs@B-N/APBA) with broad-spectrum bacterial recognition ability were synthesized for the first time to recognize and capture E. coli O157: H7 in food samples. R-CDs@BONs labeled with anti-E. coli O157:H7 monoclonal antibody (mAb@R-CDs@BONs-NH2) were used as the second recognition element to ensure the specificity for E. coli O157:H7 and form MNPs@B-N/APBA∼ E. coli O157:H7∼mAb@R-CDs@BONs-NH2 sandwich complexes, followed by releasing R-CDs to generate amplified fluorescence response signals for quantitative detection of E. coli O157:H7. The proposed method had a limit of detection with 25 CFU/mL in pure culture and contaminated lettuce samples, which the whole detection process took about 120 min. This fluorescence signal amplification biosensor has the potential to detect other pathogens in food by altering specific antibodies.

Green-Synthesized Amino Carbons for Impedimetric Biosensing of E. coli O157:H7.

This study describes the synthesis of amino-functionalized carbon nanoparticles derived from biopolymer chitosan using green synthesis and its application toward ultrasensitive electrochemical immunosensor of highly virulent Escherichia coli O157:H7 (E. coli O157:H7). The inherent advantage of high surface-to-volume ratio and enhanced rate transfer kinetics of nanoparticles is leveraged to push the limit of detection (LOD), without compromising on the selectivity. The prepared carbon nanoparticles were systematically characterized by employing CO2-thermal programmed desorption (CO2-TPD), Fourier transform infrared (FTIR) spectroscopy, X-ray photoelectron spectroscopy (XPS), ultraviolet-visible (UV-visible), and transmission electron microscopy (TEM). The estimated limit of detection of 0.74 CFU/mL and a sensitivity of 5.7 ((ΔRct/Rct)/(CFU/mL))/cm2 in the electrochemical impedance spectroscopy (EIS) affirm the utility of the sensor. The proposed biosensor displayed remarkable selectivity against interfering species, making it well suited for real-time applications. Moreover, the chitosan-derived semiconducting amino-functionalized carbon shows excellent sensitivity in a comparative analysis compared to highly conducting amine-functionalized carbon synthesized via chemical modification, demonstrating its vast potential as an E. coli sensor.

Mircrofabricating double-sided polydimethylsiloxane (PDMS) artificial phylloplane for microbial food safety research.

Leafy green surface microbiology studies often experience significant variations in results due to the heterogeneous nature of leaf surfaces. To provide a precise and controllable substitute, we microfabricated double-sided artificial leafy green phylloplanes using polydimethylsiloxane (PDMS) with a vinyl-terminated polyethylene glycol chain-based hydrophobicity modifier (PDMS-PEG) to modify PDMS hydrophobicity. We further tested the properties and applications of these artificial leaves, by examining the function of epicuticular wax, growth and survival of E. coli O157:H7 87-23 on the surface, and removal of attached E. coli cells via sanitation. The double-sided PDMS-PDMS-PEG leaves well-replicated their natural counterparts in macroscopic and microscopic structure, hydrophobicity, and E. coli O157:H7 87-23 attachment. After depositing natural epicuticular wax onto artificial leaves, the leaf surface wetting ability decreased, while E. coli O157:H7 87-23 surface retention increased. The artificial leaves supplied with lettuce lysate or bacterial growth media supported E. coli O157:H7 87-23 growth and survival similarly to those on natural leaves. In the sanitation test, the artificial lettuce leaves also displayed patterns similar to those of natural leaves regarding sanitizer efficiency. Overall, this study showcased the microfabrication and applications of double-sided PDMS-PDMS-PEG leaves as a replicable and controllable platform for future leafy green food safety studies.

Ferrous sulfate combined with ultrasound emulsified cinnamaldehyde nanoemulsion to cause ferroptosis in Escherichia coli O157:H7.

The purpose of this study was to investigate ferroptosis in Escherichia coli O157:H7 caused by ferrous sulfate (FeSO4) and to examine the synergistic effectiveness of FeSO4 combined with ultrasound-emulsified cinnamaldehyde nanoemulsion (CALNO) on inactivation of E. coli O157:H7 in vitro and in vivo. The results showed that FeSO4 could cause ferroptosis in E. coli O157:H7 via generating reactive oxygen species (ROS) and exacerbating lipid peroxidation. In addition, the results indicated that FeSO4 combined with CALNO had synergistic bactericidal effect against E. coli O157:H7 and the combined treatment could lead considerable nucleic acids and protein to release by damaging the cell membrane of E. coli O157:H7. Besides, FeSO4 combined with CALNO had a strong antibiofilm ability to inhibit E. coli O157:H7 biofilm formation by reducing the expression of genes related on biofilm formation. Finally, FeSO4 combined with CALNO exhibited the significant antibacterial activity against E. coli O157:H7 in hami melon and cherry tomato.

ß-Galactosidase-triggered in situ synthesis of yellow emitting silicon nanoparticle and its application in visual detection of E. coli O157:H7 and drug susceptibility test.

The sensitive detection of foodborne pathogenic and rapid antibiotic susceptibility testing (AST) is of great significance. This paper reports the enzyme-triggered in situ synthesis of yellow emitting silicon nanoparticles (SiNPs) and the detection of Escherichia coli (E. coli) O157:H7 in food samples and the rapid AST. The rapid counting of E. coli O157:H7 has been achieved through direct visual observation, equipment detection, and smartphone digitalization. A simple detection platform based on smartphone senses and cotton swabs has been established. Meanwhile, rapid AST based on enzyme-catalyzed SiNPs can intuitively obtain colorimetric samples. This paper established a system for bacterial enzyme-triggered in situ synthesis of SiNPs, with high responsiveness, luminescence ratio, and specificity. The detection limit for E. coli O157:H7 can reach 100 CFU/mL during 5 h, and the recovery efficiency ranges from 90.14% to 110.16%, which makes it a promising strategy for the rapid detection of E. coli O157:H7 and AST.

Characterization of diarrheagenic Escherichia coli from different cattle production systems in Brazil.

Diarrheagenic E. coli (DEC) can cause severe diarrhea and is a public health concern worldwide. Cattle are an important reservoir for this group of pathogens, and once introduced into the abattoir environment, these microorganisms can contaminate consumer products. This study aimed to characterize the distribution of DEC [Shiga toxin-producing E. coli (STEC), enteroinvasive E. coli (EIEC), enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), and enteroaggregative E. coli (EAEC)] from extensive and intensive cattle production systems in Brazil. Samples (n = 919) were collected from animal feces (n = 200), carcasses (n = 600), meat cuts (n = 90), employee feces (n = 9), and slaughterhouse water (n = 20). Virulence genes were detected by PCR in 10% of animal samples (94/919), with STEC (n = 81) as the higher prevalence, followed by EIEC (n = 8), and lastly EPEC (n = 5). Animals raised in an extensive system had a higher prevalence of STEC (average 48%, sd = 2.04) when compared to animals raised in an intensive system (23%, sd = 1.95) (Chi-square test, P < 0.001). From these animals, most STEC isolates only harbored stx2 (58%), and 7% were STEC LEE-positive isolates that were further identified as O157:H7. This study provides further evidence that cattle are potential sources of DEC, especially STEC, and that potentially pathogenic E. coli isolates are widely distributed in feces and carcasses during the slaughter process.