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1.
Food Res Int ; 184: 114252, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38609230

RESUMO

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.


Assuntos
Dimetilpolisiloxanos , Escherichia coli O157 , Meios de Cultura , Inocuidade dos Alimentos , Alface
2.
Mikrochim Acta ; 191(5): 237, 2024 04 03.
Artigo em Inglês | MEDLINE | ID: mdl-38570419

RESUMO

An ultra-sensitive fluorescent biosensor based on CDs/QDs@ZIF-8 and microfluidic fluidized bed was developed for rapid and ultra-sensitive detection of multiple target bacteria. The zeolitic imidazolate frameworks (ZIF-8) act as the carrier to encapsulate three kinds of fluorescence signal molecules from the CDs/QDs@ZIF-8 signal amplification system. Besides, three kinds of target pathogenic bacteria were automatically, continuously, and circularly captured by the magnetic nanoparticles (MNPs) in the microfluidic fluidized bed. The neutral Na2EDTA solution was the first time reported to not only dissolve the ZIF-8 frameworks from the MNPs-bacteria-CDs/QDs@ZIF-8 sandwich complexes, but also release the CDs/QDs from sandwich complexes with no loss of fluorescence signal. Due to the advantages of signal amplification and automated sample pretreatment, the proposed fluorescent biosensor can simultaneously detect Escherichia coli O157:H7, Salmonella paratyphi A, and Salmonella paratyphi B as low as 101 CFU/mL within 1.5 h, respectively. The mean recovery in spiked milk samples can reach 99.18%, verifying the applicability of this biosensor in detecting multiple bacteria in real samples.


Assuntos
Técnicas Biossensoriais , Escherichia coli O157 , Pontos Quânticos , Zeolitas , Microfluídica , Corantes
3.
Gut Microbes ; 16(1): 2331435, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38502202

RESUMO

Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is a common food-borne pathogen that can cause acute diseases. Lysine acetylation is a post-translational modification (PTM) that occurs in various prokaryotes and is regulated by CobB, the only deacetylase found in bacteria. Here, we demonstrated that CobB plays an important role in the virulence of EHEC O157:H7 and that deletion of cobB significantly decreased the intestinal colonization ability of bacteria. Using acetylation proteomic studies, we systematically identified several proteins that could be regulated by CobB in EHEC O157:H7. Among these CobB substrates, we found that acetylation at the K44 site of CesA, a chaperone for the type-III secretion system (T3SS) translocator protein EspA, weakens its binding to EspA, thereby reducing the stability of this virulence factor; this PTM ultimately attenuating the virulence of EHEC O157:H7. Furthermore, we showed that deacetylation of the K44 site, which is deacetylated by CobB, promotes the interaction between CesA and EspA, thereby increasing bacterial virulence in vitro and in animal experiments. In summary, we showed that acetylation influences the virulence of EHEC O157:H7, and uncovered the mechanism by which CobB contributes to bacterial virulence based on the regulation of CesA deacetylation.


Assuntos
Infecções por Escherichia coli , Escherichia coli O157 , Proteínas de Escherichia coli , Microbioma Gastrointestinal , Animais , Escherichia coli O157/metabolismo , Virulência , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Proteômica , Infecções por Escherichia coli/microbiologia
4.
Int J Biol Macromol ; 264(Pt 1): 130533, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38428782

RESUMO

In this work, silver­bismuth oxide encapsulated 1,3,5-triazine-bis(4-methylbenzenesulfonyl)-hydrazone functionalized chitosan (SBO/FCS) nanocomposite was synthesized by a simple hydrothermal method. The amine (-NH2) group was functionalized by the addition of cyanuric acid chloride followed by 4-methylbenzenesulfonol hydrazide. The SBO/FCS has been characterized by FT-IR, X-ray diffraction, XPS, HR-SEM, HR-TEM, AFM, and thermogravimetry (TGA). Under the optimum conditions, the SBO/FCS sensor showed brilliant electrochemical accomplishment for the sensing of glucose and H2O2 by a limit of detection (LOD) of 0.057 µM and 0.006 µM. It also showed linearity for glucose 0.008-4.848 mM and for H2O2 of 0.01-6.848 mM. Similarly, the sensor exhibited a low sensitivity to glucose (32 µA mM-1 cm-2) and a good sensitivity to H2O2 (295 µA mM-1 cm-2). In addition, that the prepared electrode could be used to sense the glucose and H2O2 levels in real samples such as blood serum and HeLa cell lines. The screen printed electrode (SPE) immunosensor could sense the E. coli O157:H7 concurrently and quantitatively with a linear range of 1.0 × 101-1.0 × 109 CFU mL-1 and a LOD of 4 CFU mL-1. Likewise, the immunosensor efficiently detect spiked E. coli O157:H7 in milk, chicken, and pork samples, with recoveries ranging from 89.70 to 104.72 %, demonstrating that the immunosensor was accurate and reliable.


Assuntos
Técnicas Biossensoriais , Bismuto , Quitosana , Escherichia coli O157 , Nanocompostos , Humanos , Peróxido de Hidrogênio/química , Prata , Glucose , Técnicas Biossensoriais/métodos , Hidrazonas , Espectroscopia de Infravermelho com Transformada de Fourier , Células HeLa , Imunoensaio/métodos , Nanocompostos/química
5.
Food Chem ; 447: 138663, 2024 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-38489878

RESUMO

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.


Assuntos
Técnicas Biossensoriais , Escherichia coli O157 , Estruturas Metalorgânicas , Transferência Ressonante de Energia de Fluorescência , Carbono , Imunoensaio , Anticorpos
6.
J Hazard Mater ; 469: 134037, 2024 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-38521032

RESUMO

Simple yet ultrasensitive and contamination-free quantification of environmental pathogenic bacteria is in high demand. In this study, we present a portable clustered regularly interspaced short palindromic repeats-associated protein 12a (CRISPR/Cas12a) powered Air-displacement enhanced Evanescent wave fluorescence Fiber-embedded microfluidic Biochip (AEFB) for the high-frequency and nucleic acid amplification-free ultrasensitive detection of Escherichia coli O157:H7. The performance of AEFB was dramatically enhanced upon employing a simple air-solution displacement process. Theoretical assays demonstrated that air-solution displacement significantly enhances evanescent wave field intensity on the fiber biosensor surface and increases the V-number in tapered fiber biosensors. Consequently, light-matter interaction is strengthened, and fluorescence coupling and collection efficiency are improved, considerably enhancing sensitivity. By integrating the CRISPR biosensing mechanism, AEFB facilitated rapid, accurate, nucleic acid amplification-free detection of E.coli O157:H7 with polymerase chain reaction (PCR)-level sensitivity (176 cfu/mL). To validate its practicality, AEFB was used to detect E.coli O157:H7 in surface water and wastewater. Comparison with RT-PCR showed a strong linear relationship (R2 = 0.9871), indicating the excellent accuracy and reliability of this technology in real applications. AEFB is highly versatile and can be easily extended to detect other pathogenic bacteria, which will significantly promote the high-frequency assessment and early-warning of bacterial contamination in aquatic environments.


Assuntos
Técnicas Biossensoriais , Escherichia coli O157 , Ácidos Nucleicos , Escherichia coli O157/genética , Sistemas CRISPR-Cas , Reprodutibilidade dos Testes , Microfluídica
7.
J Food Sci ; 89(4): 2025-2039, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38465674

RESUMO

Microbial contamination of dehydrated onion products is a challenge to the industry. The study focused on opting for a suitable drying condition for minced onion and exploring the decontamination efficacy of pulsed light (PL) treatment conditions for the dehydrated product. The minced onions were hot air dried at 55-75°C for 280 min. The drying condition selected was 195 min at 75°C with a final water activity of 0.5 and moisture content of 7% (wet basis [w.b.]). The weight losses, browning indexes (BI), shrinkage volumes (%), and thiosulfinate content were considered. The dehydrated product was exposed to PL treatment corresponding to an effective fluence range of 0.007-0.731 J/cm2. A fluence of 0.444 J/cm2 (1.8 kV for 150 s) achieved 5.00, 3.14, 2.96, and 2.98 log reduction in total plate count, yeast and mold count, Bacillus cereus 10876, and Escherichia coli ATCC 43888, respectively. The PL-treated sample (0.444 J/cm2) produced a microbially safe product with no significant difference in the moisture contents (%w.b.) and water activity (aw) from the untreated dehydrated sample. Further, a 30.9% increase in the BI and a 4.25% depletion in thiosulfinate content were observed after PL treatment. An optimum drying combination (75°C for 195 min) of minced onion followed by decontamination using pulsed light treatment at 0.444 J/cm2 fluence satisfies the microbial safety and quality. PRACTICAL APPLICATION: Dehydrated minced onion can be used for dishes requiring low water content and short cooking time. It is helpful during shortages, high price fluctuations, and famines.


Assuntos
Escherichia coli O157 , Cebolas , Microbiologia de Alimentos , Contagem de Colônia Microbiana , Descontaminação , Desidratação , Água/farmacologia , Luz
8.
J Food Prot ; 87(4): 100258, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38428462

RESUMO

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.


Assuntos
Infecções por Escherichia coli , Escherichia coli O157 , Proteínas de Escherichia coli , Escherichia coli Shiga Toxigênica , Bovinos , Animais , Infecções por Escherichia coli/veterinária , Fezes , Salmonella , Microbiologia de Alimentos
9.
ACS Appl Bio Mater ; 7(4): 2367-2377, 2024 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-38497627

RESUMO

Ensuring food safety is paramount for the food industry and global health concerns. In this study, we have developed a method for the detection of prevalent foodborne pathogenic bacteria, including Escherichia coli, Salmonella spp., Listeria spp., Shigella spp., Campylobacter spp., Clostridium spp., and Vibrio spp., utilizing antibody-aptamer arrays. To enhance the fluorescence signals on the microarray, the mesoporous silica nanoparticles (MSNs) conjugated with fluorescein, streptavidin, and seven detection antibodies-biotin were employed, forming fluorescein doped mesoporous silica nanoparticles conjugated with detection antibodies (MSNs-Flu-SA-Abs) complexes. The array pattern was designed for easy readability and enabled the simultaneous detection of all seven foodborne pathogens, referred to as the 7FP-biochip. Following the optimization of MSNs-Flu-SA-Abs complexes attachment and enhancement of the detection signal in fluorescent immunoassays, a high level of sensitivity was achieved. The detection limits for the seven pathogens in both buffer and food samples were 102 CFU/mL through visual screening, with fluorescent intensity quantification achieving levels as low as 20-34 CFU/g were achieved on the antibody-aptamer arrays. Our antibody-aptamer array offers several advantages, including significantly reduced nonspecific binding with no cross-reaction between bacteria. Importantly, our platform detection exhibited no cross-reactivity among the tested bacteria in this study. The multiplex detection of foodborne pathogens in canned tuna samples with spiked bacteria was successfully demonstrated in real food measurements. In conclusion, our study presents a promising method for detecting multiple foodborne pathogens simultaneously. With its high sensitivity and specificity, the developed antibody-aptamer array holds great potential for enhancing food safety and public health.


Assuntos
Escherichia coli O157 , Nanopartículas , Contaminação de Alimentos/análise , Microbiologia de Alimentos , Bactérias , Fluoresceínas , Tecnologia
10.
Analyst ; 149(8): 2436-2444, 2024 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-38498083

RESUMO

Paper-based electrochemical sensors have the characteristics of flexibility, biocompatibility, environmental protection, low cost, wide availability, and hydropathy, which make them very suitable for the development and application of biological detection. This work proposes electrospun cellulose acetate nanofiber (CA NF)-decorated paper-based screen-printed (PBSP) electrode electrochemical sensors. The CA NFs were directly collected on the PBSP electrode through an electrospinning technique at an optimized voltage of 16 kV for 10 min. The sensor was functionalized with different bio-sensitive materials for detecting different targets, and its sensing capability was evaluated by CV, DPV, and chronoamperometry methods. The test results demonstrated that the CA NFs enhanced the detection sensitivity of the PBSP electrode, and the sensor showed good stability, repeatability, and specificity (p < 0.01, N = 3). The electrochemical sensing of the CA NF-decorated PBSP electrode exhibited a short detection duration of ∼5-7 min and detection ranges of 1 nmol mL-1-100 µmol mL-1, 100 fg mL-1-10 µg mL-1, and 1.5 × 102-106 CFU mL-1 and limits of detection of 0.71 nmol mL-1, 89.1 fg mL-1, and 30 CFU mL-1 for glucose, Ag85B protein, and E. coli O157:H7, respectively. These CA NF-decorated PBSP sensors can be used as a general electrochemical tool to detect, for example, organic substances, proteins, and bacteria, which are expected to achieve point-of-care testing of pathogenic microorganisms and have wide application prospects in biomedicine, clinical diagnosis, environmental monitoring, and food safety.


Assuntos
Técnicas Biossensoriais , Celulose/análogos & derivados , Escherichia coli O157 , Nanofibras , Nanofibras/química , Celulose/química , Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas/métodos
11.
Anal Chem ; 96(13): 5205-5214, 2024 Apr 02.
Artigo em Inglês | MEDLINE | ID: mdl-38481140

RESUMO

Pathogenic diseases that trigger food safety remain a noteworthy concern due to substantial public health, economic, and social burdens worldwide. It is vital for developing an integrated diagnosis and treatment strategy for bacteria, which could achieve quick detection of pathogenic bacteria and the inhibition of multidrug-resistant bacteria. Herein, we reported an organic molecule (M-3) possessed strong light capture capacity, emerging a low energy gap and ΔEST. Subsequently, M-3 was integrated into a nanostructured system (BTBNPs) with excellent ROS generation, light absorption capability, and photothermal performance. Reactive oxygen species (ROS) generated by BTBNPs were mainly free radicals from a type I mechanism, and the high photothermal conversion efficiency of BTBNPs was 41.26%. Benefiting from these advantages of BTBNPs, BTBNPs could achieve a ∼99% antibacterial effect for Escherichia coli O157:H7 with 20 µM dosage and 5 min of irradiation. Furthermore, the limit of detection (LoD) of the proposed BTBNPs-LFIA (colorimetric and photothermal modalities) for detecting E. coli O157:H7 was 4105 and 419 CFU mL-1, respectively. Overall, this work is expected to provide a new and sophisticated perspective for integrated diagnosis and treatment systems regarding pathogenic bacteria.


Assuntos
Escherichia coli O157 , Nanopartículas Multifuncionais , Microbiologia de Alimentos , Espécies Reativas de Oxigênio , Limite de Detecção
12.
Anal Chem ; 96(13): 5340-5347, 2024 Apr 02.
Artigo em Inglês | MEDLINE | ID: mdl-38501977

RESUMO

Fully integrated devices that enable full functioning execution without or with minimum external accessories or equipment are deemed to be one of the most desirable and ultimate objectives for modern device design and construction. Escherichia coli O157:H7 (E. coli O157:H7) is often linked to outbreaks caused by contaminated water and food. However, the sensors that are currently used for point-of-care E. coli O157:H7 (E. coli O157:H7) detection are often large and cumbersome. Herein, we demonstrate the first example of a handheld and pump-free fully integrated electrochemical sensing platform with the capability to point-of-care test E. coli O157:H7 in the actual samples of E. coli O157:H7-spiked tap water and E. coli O157:H7-spiked watermelon juice. This platform was made possible by overcoming major engineering challenges in the seamless integration of a microfluidic module for pump-free liquid sample collection and transportation, a sensing module for efficient E. coli O157:H7 testing, and an electronic module for automatically converting and wirelessly transmitting signals into a single and compact electrochemical sensing platform that retains its inimitable stand-alone, handheld, pump-free, and cost-effective feature. Although our primary emphasis in this study is on detecting E. coli O157:H7, this pump-free fully integrated handheld electrochemical sensing platform may also be used to monitor other pathogens in food and water by including specific antipathogen antibodies.


Assuntos
Escherichia coli O157 , Anticorpos , Testes Imediatos , Sistemas Automatizados de Assistência Junto ao Leito , Água , Microbiologia de Alimentos
13.
Nature ; 628(8006): 180-185, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38480886

RESUMO

The gut microbiome has major roles in modulating host physiology. One such function is colonization resistance, or the ability of the microbial collective to protect the host against enteric pathogens1-3, including enterohaemorrhagic Escherichia coli (EHEC) serotype O157:H7, an attaching and effacing (AE) food-borne pathogen that causes severe gastroenteritis, enterocolitis, bloody diarrhea and acute renal failure4,5 (haemolytic uremic syndrome). Although gut microorganisms can provide colonization resistance by outcompeting some pathogens or modulating host defence provided by the gut barrier and intestinal immune cells6,7, this phenomenon remains poorly understood. Here, we show that activation of the neurotransmitter receptor dopamine receptor D2 (DRD2) in the intestinal epithelium by gut microbial metabolites produced upon dietary supplementation with the essential amino acid L-tryptophan protects the host against Citrobacter rodentium, a mouse AE pathogen that is widely used as a model for EHEC infection8,9. We further find that DRD2 activation by these tryptophan-derived metabolites decreases expression of a host actin regulatory protein involved in C. rodentium and EHEC attachment to the gut epithelium via formation of actin pedestals. Our results reveal a noncanonical colonization resistance pathway against AE pathogens that features an unconventional role for DRD2 outside the nervous system in controlling actin cytoskeletal organization in the gut epithelium. Our findings may inspire prophylactic and therapeutic approaches targeting DRD2 with dietary or pharmacological interventions to improve gut health and treat gastrointestinal infections, which afflict millions globally.


Assuntos
Citrobacter rodentium , Mucosa Intestinal , Receptores de Dopamina D2 , Triptofano , Animais , Feminino , Humanos , Masculino , Camundongos , Citoesqueleto de Actina/efeitos dos fármacos , Citoesqueleto de Actina/metabolismo , Actinas/metabolismo , Carga Bacteriana/efeitos dos fármacos , Citrobacter rodentium/crescimento & desenvolvimento , Citrobacter rodentium/metabolismo , Citrobacter rodentium/patogenicidade , Suplementos Nutricionais , Modelos Animais de Doenças , Infecções por Enterobacteriaceae/microbiologia , Infecções por Enterobacteriaceae/prevenção & controle , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/prevenção & controle , Escherichia coli O157/patogenicidade , Escherichia coli O157/fisiologia , Mucosa Intestinal/citologia , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiologia , Receptores de Dopamina D2/metabolismo , Triptofano/administração & dosagem , Triptofano/metabolismo , Triptofano/farmacologia
14.
Biosensors (Basel) ; 14(3)2024 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-38534259

RESUMO

Accurate sample heating is vital for nucleic acid extraction and amplification, requiring a sophisticated thermal cycling process in nucleic acid detection. Traditional molecular detection systems with heating capability are bulky, expensive, and primarily designed for lab settings. Consequently, their use is limited where lab systems are unavailable. This study introduces a technique for performing the heating process required in molecular diagnostics applicable for point-of-care testing (POCT), by presenting a method for crafting customized heaters using freely patterned nichrome (NiCr) wire. This technique, fabricating heaters by arranging protrusions on a carbon black-polydimethylsiloxane (PDMS) cast and patterning NiCr wire, utilizes cost-effective materials and is not constrained by shape, thereby enabling customized fabrication in both two-dimensional (2D) and three-dimensional (3D). To illustrate its versatility and practicality, a 2D heater with three temperature zones was developed for a portable device capable of automatic thermocycling for polymerase chain reaction (PCR) to detect Escherichia coli (E. coli) O157:H7 pathogen DNA. Furthermore, the detection of the same pathogen was demonstrated using a customized 3D heater surrounding a microtube for loop-mediated isothermal amplification (LAMP). Successful DNA amplification using the proposed heater suggests that the heating technique introduced in this study can be effectively applied to POCT.


Assuntos
Ligas de Cromo , Escherichia coli O157 , Ácidos Nucleicos , Patologia Molecular , Técnicas de Amplificação de Ácido Nucleico/métodos , DNA , Técnicas de Diagnóstico Molecular/métodos
15.
Sci Rep ; 14(1): 5821, 2024 03 09.
Artigo em Inglês | MEDLINE | ID: mdl-38461188

RESUMO

The aim of this study was to compare Illumina and Oxford Nanopore Technology (ONT) sequencing data to quantify genetic variation to assess within-outbreak strain relatedness and characterise microevolutionary events in the accessory genomes of a cluster of 23 genetically and epidemiologically linked isolates related to an outbreak of Shiga toxin-producing Escherichia coli O157:H7 caused by the consumption of raw drinking milk. There were seven discrepant variants called between the two technologies, five were false-negative or false-positive variants in the Illumina data and two were false-negative calls in ONT data. After masking horizontally acquired sequences such as prophages, analysis of both short and long-read sequences revealed the 20 isolates linked to the outbreak in 2017 had a maximum SNP distance of one SNP between each other, and a maximum of five SNPs when including three additional strains identified in 2019. Analysis of the ONT data revealed a 47 kbp deletion event in a terminal compound prophage within one sample relative to the remaining samples, and a 0.65 Mbp large chromosomal rearrangement (inversion), within one sample relative to the remaining samples. Furthermore, we detected two bacteriophages encoding the highly pathogenic Shiga toxin (Stx) subtype, Stx2a. One was typical of Stx2a-phage in this sub-lineage (Ic), the other was atypical and inserted into a site usually occupied by Stx2c-encoding phage. Finally, we observed an increase in the size of the pO157 IncFIB plasmid (1.6 kbp) in isolates from 2019 compared to those from 2017, due to the duplication of insertion elements within the plasmids from the more recently isolated strains. The ability to characterize the accessory genome in this way is the first step to understanding the significance of these microevolutionary events and their impact on the genome plasticity and virulence between strains of this zoonotic, foodborne pathogen.


Assuntos
Bacteriófagos , Infecções por Escherichia coli , Escherichia coli O157 , Sequenciamento por Nanoporos , Humanos , Animais , Leite , Toxina Shiga/genética , Bacteriófagos/genética , Prófagos/genética , Surtos de Doenças , Infecções por Escherichia coli/epidemiologia
16.
Microb Biotechnol ; 17(3): e14437, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38465735

RESUMO

Escherichia coli O157:H7 causes >73,000 foodborne illnesses in the United States annually, many of which have been associated with fresh ready-to-eat produce including cantaloupe melons. In this study, we created a produce-associated bacterial (PAB) library containing >7500 isolates and screened them for the ability to inhibit the growth of E. coli O157:H7 using an in vitro fluorescence-based growth assay. One isolate, identified by 16S and whole-genome sequence analysis as Enterobacter asburiae, was able to inhibit the growth of E. coli by ~30-fold in vitro and produced zones of inhibition between 13 and 21 mm against 12 E. coli outbreak strains in an agar spot assay. We demonstrated that E. asburiae AEB30 was able to grow, persist and inhibit the growth of E. coli on cantaloupe melons under simulated pre- and post-harvest conditions. Analysis of the E. asburiae AEB30 genome revealed an operon encoding a contact-dependent growth inhibition (CDI) system that when mutated resulted in the loss of E. coli growth inhibition. These data suggest that E. asburiae AEB30 is a potential biocontrol agent to prevent E. coli contamination of cantaloupe melons in both pre- and post-harvest environments and that its mode of action is via a CDI system.


Assuntos
Cucumis melo , Cucurbitaceae , Enterobacter , Escherichia coli O157 , Microbiologia de Alimentos , Cucumis melo/microbiologia , Cucurbitaceae/microbiologia , Contagem de Colônia Microbiana
17.
Int J Food Microbiol ; 413: 110609, 2024 Mar 02.
Artigo em Inglês | MEDLINE | ID: mdl-38330783

RESUMO

Falafel is a popular breakfast food in the Middle East that has been recently involved in several outbreaks of foodborne illnesses. The aim of the study was to explore the growth behavior of Salmonella enterica, Escherichia coli O157:H7, Shigella sonnie, Shigella flexneri, Listeria monocytogenes and Staphylococcus aureus in falafel paste (FP) under different storage temperatures (4, 10, or 24 °C) for 14 days. FP (pH = 6.2, aw = 0.96) was inoculated with 5.0 to 6.0 log CFU/g of each of the pathogens separately. Salmonella spp. significantly declined by 1.5 log at 4 °C but grew significantly by ca. 2 and 4 log at 10 and 24 °C, respectively after 14 days. E. coli O157:H7 significantly increased (4.5 log) in FP when stored under 24 °C and survived at a level of ~105 CFU/g at 10 °C. Comparatively, Sh. sonnie and Sh. flexneri showed a better survival pattern in FP stored under 4 °C and grew (˃ 3 log) after 5 days at 10 and 24 °C. L. monocytogenes was capable of growing by 1.9 and 4.3 log after 14 d days and by 3.9 log after 3 days at 4, 10, or 24 °C, respectively. No significant decline in S. aureus counts at 4 and 10 °C occurred, however, it increased significantly to ˃ 7 log CFU/g at 24 °C. Total mesophilic count and yeast and mold count reached to spoilage levels (˃107 CFU/g) in un-inoculated FP after 1 and 3 days of storage at 24 and 10 °C, respectively. FP could support the growth of common foodborne pathogens and hence it is recommended to utilize natural antimicrobials in FP and keep the product under refrigeration (4 °C) to preclude the growth of vegetative foodborne pathogens.


Assuntos
Escherichia coli O157 , Listeria monocytogenes , Temperatura , Staphylococcus aureus , Microbiologia de Alimentos , Verduras , Contagem de Colônia Microbiana
18.
Prev Vet Med ; 224: 106132, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38335831

RESUMO

Some sorbitol non-fermenting E. coli (SN-F E. coli) and all E. coli O157 are zoonotic. Contamination of beef with zoonotic E. coli at the processing or retail point is a significant public health problem. Despite the public health importance of these organisms, there is no published data on the prevalence and antimicrobial resistance (AMR) of zoonotic E. coli from Delta State, Nigeria. Consequently, this study determined the prevalence and AMR of SN-F E. coli and E. coli O157 isolates from meat contact surfaces at the processing and retail points in the study area. The isolation, biochemical and serological characterisations and AMR status of the isolates were performed following standard microbiological methods. Overall prevalence of SN-F E. coli and E. coli O157 were 13.8% (56/406) and 1.5% (6/406), respectively. Majority of the 56 SN-F E. coli (64.3%, 36/56) and all the six E. coli O157 (10.7%, 6/56) detected in this study were found at the meat processing points. Most of the SN-F E. coli were isolated at the slaughterhouse floor (31%), meat hooks (17.2%) and meat sellers' knives (17.2%). The SN-F E. coli exhibited greater AMR to ampicillin (67.9%), gentamycin (64.3%) and tetracycline (50%) than other antimicrobial agents tested. No isolate was resistant to aztreonam. All six E. coli O157 isolates were resistant to enrofloxacin. Overall, 23 AMR patterns, comprised 14 from meat processing points and nine from meat retailing points, were observed from the 56 antimicrobial-resistant SN-F E. coli isolates. All the six E. coli O157 and 73.2% (41/56) of the SN-F E. coli isolates were multidrug-resistant. An overall mean multiple antimicrobial resistance index of 0.6 was recorded. Multidrug-resistant zoonotic E. coli were detected at meat processing and retail points in Delta State, Nigeria. The findings warrant the adoption of One Health control approach, "farm to fork" principle of food safety and prudent use of antimicrobial agents in animal agriculture. These may help to limit beef contamination with multidrug-resistant zoonotic E. coli at the processing and retailing points, for public health safety.


Assuntos
Anti-Infecciosos , Escherichia coli O157 , Bovinos , Animais , Saúde Pública , Nigéria/epidemiologia , Antibacterianos/farmacologia , Carne/microbiologia , Farmacorresistência Bacteriana , Testes de Sensibilidade Microbiana/veterinária
19.
Virulence ; 15(1): 2313407, 2024 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-38357901

RESUMO

Enterohemorrhagic Escherichia coli (EHEC) is an important zoonotic pathogen that is a major cause of foodborne diseases in most developed and developing countries and can cause uncomplicated diarrhoea, haemorrhagic colitis, and haemolytic uraemic syndrome. O islands (OIs), which are unique genomic islands in EHEC O157:H7, are composed of 177 isolated genomic features and harbour 26% of the total genes that are absent in the non-pathogenic E. coli K-12 genome. In the last twenty years, many OI-encoded proteins have been characterized, including proteins regulating virulence, motility, and acid resistance. Given the critical role of regulatory proteins in the systematic and hierarchical regulation of bacterial biological processes, this review summarizes the OI-encoded regulatory proteins in EHEC O157:H7 characterized to date, emphasizing OI-encoded regulatory proteins for bacterial virulence, motility, and acid resistance. This summary will be significant for further exploration and understanding of the virulence and pathogenesis of EHEC O157:H7.


Assuntos
Escherichia coli Êntero-Hemorrágica , Infecções por Escherichia coli , Escherichia coli O157 , Proteínas de Escherichia coli , Humanos , Ilhas Genômicas , Escherichia coli O157/genética , Fatores de Transcrição/genética , Escherichia coli Êntero-Hemorrágica/genética , Virulência/genética , Infecções por Escherichia coli/microbiologia , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo
20.
Food Chem ; 443: 138564, 2024 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-38320374

RESUMO

Escherichia coli O157:H7 (E. coli O157:H7) emerges as a significantly worrisome pathogen associated with foodborne illnesses, emphasizing the imperative for creating precise detection tools. In this investigation, we developed a sensitive colorimetric biosensor for detecting E. coli O157:H7. It was constructed using a nanozyme comprised of Au@Fe3O4 NPs, which was fabricated and subsequently modified with an aptamer (Apt). The nanozyme harnesses its inherent peroxidase-like activity to facilitate the transformation of reduced TMB into its oxidized form in the presence of H2O2, resulting in a noticeable shift to a blue color. However, the presence of E. coli O157:H7 effectively diminished the absorbance of oxidized TMB. Consequently, the normalized absorbance at 652 nm demonstrates a linear decrease corresponding to concentrations of E. coli O157:H7 within the range of 101 to 108 CFU mL-1 with a low limit of detection (LOD, S/N = 3) of 3 CFU mL-1.


Assuntos
Técnicas Biossensoriais , Escherichia coli O157 , Colorimetria , Peróxido de Hidrogênio , Peroxidases , Técnicas Biossensoriais/métodos , Microbiologia de Alimentos
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