Coli O157 Research Articles

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.

Effect of Ultrasonic Extract of Capparis spinosa‎‎ Fruits Against E. coli‎‎ O157:H7‎


 
 
 
 E. coli‎‎ O157:H7, is one of the main causes of diarrhea and the most prevalent bacterial ‎infection that causes serious illnesses. This research was conducted to investigate the effect of Capparis ‎spinosa fruit ultrasonic extract against resistant E. coli‎‎ O157:H7 was isolated from stools of calves ‎that were suffering from diarrhea; the bacteria were identified by the Vitek 2 system and a latex agglutination ‎test. C. spinosa‎‎‎ was extracted by ultrasonic waves water bath. The phytochemicals were carried out on C. ‎spinosa fruit extract to detect the secondary metabolites. The MIC of the extract was calculated with ‎concentrations of 400, 800, 1600, 3200, 6400, 12800 and 25600 51200 µg/mL by microdilution method ‎‎(checkerboard). While using a field-emission scanning electron microscope to observe the morphological ‎alterations in E. coli‎‎ O157:H7 sample. The findings of this study revealed that the extract contains some ‎biologically active compounds like alkaloids, flavonoids, steroids, glycosides, tannins, cumarines, saponins, ‎quinones, and amino acids. That extract of C. spinosa‎‎‎ had a MIC of 6400 µg/mL and had a perfect action ‎against E. coli‎‎. O157:H7 by forming vacuoles within the cells and that internal content had seeped out as pore ‎formation. This finding could potentially provide an explanation for the traditional utilization of ‎this plant material as an antibacterial agent‎‎.
 
 
 

Microbiological Evaluation of Different Types of Branded and Non-branded Ready-to-Eat Snacks Sold in Elementary Schools of District Peshawar, Pakistan

The lack of hygiene in the preparation and packing of ready-to-eat foods is fast becoming a serious public health concern, especially for school children. The intervention of different pathogenic microorganisms in these foods poses the risk of foodborne disease outbreaks. The current study was designed to assess bacterial contamination in different types of ready-to-eat (branded and non-branded) snacks purchased from September 2021 to December 2021 in various elementary schools of Peshawar, Pakistan. A total of 20 samples were collected and analyzed using the pour plate method for total plate count (TPC). Moreover, the multiple fermentation tube method was used for total coliforms (TC) and fecal coliforms (FC). Escherichia coli isolates were identified using E. coli O157:H7 latex test reagent kit Pro Lab, Canada. The results revealed contamination by TPC (6.67%), TC (40%), FC (33.33%), and E. coli (20%) in the samples. These values were higher than the permissible limits set by Food and Agriculture Organization (FAO). Hence, it was concluded that there is a practice gap in food safety knowledge among ready-to-eat food vendors. The vendors are usually untrained and lack the knowledge of proper hygiene and food handling procedures. It is suggested here that the government should pay special attention towards improving public awareness regarding food safety and quality of ready-to-eat foods sold in Peshawar, Pakistan.

Epinecidin-1 (an active marine antimicrobial peptide): Effects on the survival of inoculated Escherichia Coli O157:H7 And Staphylococcus aureus bacteria, antioxidant, and sensory attributes in raw milk.

This study aimed to evaluate the effects of Epinecidin-1 (Epi-1) on total viable count (TVC), total psychrotrophic count (TPC), sensory attributes, and the survival of Escherichia Coli O157:H7 and Staphylococcus aureus bacteria inoculated in pasteurized milk samples during cold storage (4°C). Four treatments of milk samples were prepared including milk samples containing three concentrations of Epi-1 (0.0025, 0.005, and 0.01%) and control (without Epi-1). The treated milk samples were evaluated in vitro (minimum inhibitory concentration, Minimum bactericidal concentration, disk diffusion test, DPPH, reducing power assays) and in vivo (TVC, TPC, sensory properties, and enumeration of inoculated E. coli and S. aureus) during 9 days at cold storage. The best antibacterial and antioxidant power of Epi-1 was observed at a concentration of 0.01%. Based on the MICs and MBCs, the most susceptible and resistant bacteria to Epi-1 were B. cereus and S. aureus strains, respectively. The DPPH scavenging potential of Epi-1 was in the range of 77%-80%. Treated samples containing 0.01% Epi-1 had the lowest TVC and TPC and reached 3.9 and 2.96 CFU log/mL at the end day of storage. A decrease of 6 and 1.4 logs CFU/g of E. coli O157:H7 and S. aureus was seen in all treatments containing Epi-1, respectively, on the last day of storage period. There are no unpleasant sensory properties in treated samples with Epi-1. Our results indicate that Epi-1 has good potential as a bio-preservative to prevent raw milk spoilage and reduction of milk-borne pathogens.

The Use of Phage Cocktail and Various Antibacterial Agents in Combination to Prevent the Emergence of Phage Resistance.

Bacterial food poisoning cases due to Salmonella and E. coli O157:H7 have been linked with the consumption of a variety of food products, threatening public health around the world. This study describes the combined effects of a phage cocktail (STG2, SEG5, and PS5), EDTA, nisin, and polylysine against the bacterial cocktail consisting of S. typhimurium, S. enteritidis, and E. coli O157:H7. Overall, phage cocktail (alone or in combination with nisin or/and polylysine) not only showed great antibacterial effects against bacterial cocktail at different temperatures (4 °C, 24 °C, and 37 °C), but also totally inhibited the emergence of phage resistance during the incubation period. These results suggest that the combination of phages with nisin or/and polylysine has great potential to simultaneously control S. typhimurium, S. enteritidis, and E. coli O157:H7.

Anti-viral, anti-bacterial, but non-cytotoxic nanocoating for reusable face mask with efficient filtration, breathability, and robustness in humid environment

We fabricated a multi-use face mask with anti-bacterial and anti-viral, but non-cytotoxic properties by adding a functional nanocoating on an electrospun nanofiber filter. The nanocoating consists of a copolymer synthesized from dimethyaminomethyl styrene (DMAMS) monomer containing anti-microbial tertiary amine moiety, and a biocompatible crosslinker, ethylene glycol dimethylacrylate (EGDMA). Initiated chemical vapor deposition process is utilized to deposit the nanocoating in order to impart desired functionalities conformally onto the nanofiber filter without deforming its highly porous structure. The synthesized non-cytotoxic copolymer of p(DMAMS-co-EGDMA), or pDE film showed excellent biocidal efficacy of > 99.995% and > 99.985% against Escherichia coli O157: H7 (E. coli O157: H7) and Staphylococcus aureus, respectively. The pDE-coated nanofiber filter shows a filtration efficiency of 91% with extremely low air resistance of ∼ 50 Pa. In highly humid conditions, the pDE-coated nanofiber filter shows long-term anti-microbial efficacy of 99.89% for E. coli O157: H7 up to 168 h, 99.21%, and 99.23% for human influenza virus (H1N1) strain A/Puerto Rico/8/1934 and human beta-coronavirus strain OC43 up to 100 h, without sacrificing breathability, and filtration performance. Furthermore, the surface-modified filter can easily eliminate dead pathogens with a simple water rinse, which enabled the full retention of the filtration efficiency even in six washing cycles, proving excellent reusability. We envision the highly breathable, antimicrobial, multi-use face mask will serve as an environmentally free platform for personal protective equipment against future viral threats.

Simultaneous and rapid screening of live and dead E. coli O157:H7 with three signal outputs: An all-in-one biosensor using phage-apoferritin@CuO2 signal tags on MXenes-modified electrode platform

Simultaneous detection of live and dead bacteria is a huge challenge for food safety. To solve this issue, an all-in-one biosensor for bacteria was developed using the phage-apoferritin@CuO2 (phage-Apo@CP) probe on an antimicrobial peptide (AMP)/MXenes-modified detection platform. With the specific recognition of AMP and phage-Apo@CP, the biosensor for the target Escherichia coli O157:H7 (E. coli O157:H7) presented multi-mode (bioluminescent, colorimetric, and electrochemical) signals to simultaneously measure live and dead bacteria. The bioluminescent signal caused by the adenosine triphosphate (ATP) from the bacteria was used to quantify live bacteria. The colorimetric and voltammetric signals triggered by ·OH and Cu2+ from the probe with the assistance of acid could rapidly screen and quantitative determination of total E. coli O157:H7 concentration. Thus, the dead one was obtained according to the total and live ones. All three signals could be mutually corrected to improve the accuracy. The biosensor was successfully used for on-site measurement of live and dead E. coli O157:H7 in food samples with the limit of detection of 30 CFU/mL for live ones and 6 CFU/mL for total bacteria within 50 min. This work presents a novel pathway for rapid and simultaneous quantification of both live and dead bacteria.

Ultrasensitive and Rapid Visual Detection of Escherichia coli O157:H7 Based on RAA-CRISPR/Cas12a System.

Escherichia coli (E. coli) O157:H7 is a major foodborne and waterborne pathogen that can threaten human health. Due to its high toxicity at low concentrations, it is crucial to establish a time-saving and highly sensitive in situ detection method. Herein, we developed a rapid, ultrasensitive, and visualized method for detecting E. coli O157:H7 based on a combination of Recombinase-Aided Amplification (RAA) and CRISPR/Cas12a technology. The CRISPR/Cas12a-based system was pre-amplified using the RAA method, which showed high sensitivity and enabled detecting as low as ~1 CFU/mL (fluorescence method) and 1 × 102 CFU/mL (lateral flow assay) of E. coli O157:H7, which was much lower than the detection limit of the traditional real-time PCR technology (103 CFU/mL) and ELISA (104~107 CFU/mL). In addition, we demonstrated that this method still has good applicability in practical samples by simulating the detection in real milk and drinking water samples. Importantly, our RAA-CRISPR/Cas12a detection system could complete the overall process (including extraction, amplification, and detection) within 55 min under optimized conditions, which is faster than most other reported sensors, which take several hours to several days. The signal readout could also be visualized by fluorescence generated with a handheld UV lamp or a naked-eye-detected lateral flow assay depending on the DNA reporters used. Because of the advantages of being fast, having high sensitivity, and not requiring sophisticated equipment, this method has a promising application prospect for in situ detection of trace amounts of pathogens.

Barotolerance of Acid-adapted and Cold-adapted Bacterial Isolates of E. coli O157:H7, Salmonella spp., and L. monocytogenes in an Acidic Buffer Model

The fruit and vegetable juice industry has shown a growing trend in minimally processed juices. A frequent technology used in the production of functional juices is cold pressure, which refers to the application of high pressure processing (HPP) at low temperatures to inactivate foodborne pathogens. HPP juice manufacturers are required to demonstrate a 5-log reduction of the pertinent microorganism to comply with FDA Juice HACCP. However, there is no consensus on validation study approaches for bacterial strain selection or their preparation. Individual bacterial strains were grown using three different growth conditions: neutral, cold-adapted, and acid-adapted. Approximately 6.0–7.0 log CFU/mL of the matrix-adapted bacterial strains were inoculated individually into buffered peptone water (BPW) at pH 3.50 ± 0.10 (HCl adjusted) and treated at sublethal pressures of 500 MPa for Escherichia coli O157:H7 and 200 MPa for Salmonella spp. and Listeria monocytogenes (180 s, 4°C). Analyses were conducted at 0, 24, and 48 h (4°C storage) post-HPP on nonselective media. E. coli O157:H7 exhibited greater barotolerance than Salmonella spp. and L. monocytogenes. In neutral growth conditions, E. coli O157:H7 strain TW14359 demonstrated the greatest resistance (2.94 ± 0.64 log reduction), and E. coli O157:H7 strain SEA13B88 was significantly more sensitive (P < 0.05). Salmonella isolates, neutral and acid-adapted, expressed similar barotolerance to one another. Cold-adapted S. Cubana and S. Montevideo showed greater resistance compared to other cold-adapted strains. Acid-adapted L. monocytogenes strain MAD328 had <1.00 ± 0.23 log reduction while acid-adapted L. monocytogenes strains CDC and Scott A were significantly more sensitive (P < 0.05) with reductions of 2.13 ± 0.48 and 3.43 ± 0.50 log CFU/mL, respectively. These results suggested, under the conditions tested, bacterial strain and preparation methods influence HPP efficacy and should be considered when conducting validation studies.

Biological control of foodborne pathogens by lactic acid bacteria: A focus on juice processing industries

The use of lactic acid bacteria (LAB) in foods as biocontrol agents against foodborne pathogens has become increasingly known. Under the premise that controlling the adhesion of microorganisms to food contact surfaces is an essential step for meeting the goals of food processing, the aim of this work was to investigate the inhibitory and anti-biofilm effectiveness of Lactobacillus rhamnosus GG (ATCC 53103) and Lactobacillus casei (ATCC 393) against Escherichia coli O157:H7, Salmonella enterica and Listeria monocytogenes. Lactobacillus strains (108CFU/ml) and pathogens (104CFU/ml) were evaluated to monitor LAB anti-adhesive and antibiofilm effect, in two main scenarios: (i) co-adhesion and (ii) pathogen incorporation to stainless steel surfaces with a protective biofilm of Lactobacillus cells. In (i) the predominant effect was observed in L. rhamnosus against S. enterica and L. monocytogenes, whereas in (ii) both LAB significantly reduced the number of pathogenic adherent cells. The effect of pre-established LAB biofilms was more successful in displacing the three pathogens than when they were evaluated under co-adhesion. These findings show that both LAB can be considered good candidates to prevent or inhibit the adhesion and colonization of L. monocytogenes, S. enterica and E. coli O157:H7 on surfaces and conditions of relevance for juice processing industries, offering alternatives for improving the safety and quality of fruit-based products.

The potential risk associated with foodborne pathogens in a watershed: E. Coli O157:H7 in dairy cattle.

We carried out an investigation to determine the occurrence of E. coli O157:H7 in dairy herds in the Delaware County watershed and to identify the factors that play a role in the likelihood of presence of this organism among animals on these farms. The pathogen poses risk of environmental degradation and health to the inhabitants. A total of 2,162 fecal samples were collected per rectum from a representative sample of cattle on 27 dairy farms. The samples were investigated for the presence of E. coli O157:H by initially enriching using a bacteriological media and detection of the pathogen using real-time PCR technique. E. coli O157:H7 was detected in 74% of the herds in the target population and in 3.7% of samples collected. An additional 54 animals were identified that were infected with O157 non-H7 strains of E. coli among 15 farms. Several putative risk factors were associated with the detection of the pathogen on the enrolled farms included age, housing calves indoors, group housing for calves, housing calves in the calf barn, presence of dogs on the farm, and housing post-weaned calves in a cow barn or heifer barn versus a greenhouse. In conclusion, E. coli O157:H7 was present on the dairy farms of Delaware County and may pose a threat to the people that live and work there. The risk associated with the detection of this pathogen could be reduced by adjusting management factors that were identified in this study. This article is protected by copyright. All rights reserved.

The role of PhoP/PhoQ system in regulating stress adaptation response in Escherichia coli O157:H7.

The development of acid tolerance response (ATR) as a result of low pH in Escherichia coli O157:H7 (E. coli O157:H7) contaminating beef during processing is considered a major food safety concern. Thus, in order to explore the formation and molecular mechanisms of the tolerance response of E. coli O157:H7 in a simulated beef processing environment, the resistance of a wild-type (WT) strain and its corresponding ΔphoP mutant to acid, heat, and osmotic pressure was evaluated. Strains were pre-adapted under different conditions of pH (5.4 and 7.0), temperature (37°C and 10°C), and culture medium (meat extract and Luria-Bertani broth media). In addition, the expression of genes related to stress response and virulence was also investigated among WT and ΔphoP strains under the tested conditions. Pre-acid adaptation increased the resistance of E. coli O157:H7 to acid and heat treatment while resistance to osmotic pressure decreased. Moreover, acid adaptation in meat extract medium simulating slaughter environment increased ATR, whereas pre-adaptation at 10°C reduced the ATR. Furthermore, it was shown that mildly acidic conditions (pH=5.4) and the PhoP/PhoQ two-component system (TCS) acted synergistically to enhance acid and heat tolerance in E. coli O157:H7. Additionally, the expression of genes related to arginine and lysine metabolism, heat shock, and invasiveness was up-regulated, which revealed that the mechanism of acid resistance and cross-protection under mildly acidic conditions was mediated by the PhoP/PhoQ TCS. Both acid adaptation and phoP gene knockout reduced the relative expression of stx1 and stx2 genes which were considered as critical pathogenic factors. Collectively, the current findings indicated that ATR could occur in E. coli O157:H7 during beef processing. Thus, there is an increased food safety risk due to the persistence of tolerance response in the following processing conditions. The present study provides a more comprehensive basis for the effective application of hurdle technology in beef processing.

Isolation, Characterization, and Comparative Genomic Analysis of Bacteriophage Ec_MI-02 from Pigeon Feces Infecting Escherichia coli O157:H7.

The most significant serotype of Shiga-toxigenic Escherichia coli that causes foodborne illnesses is Escherichia coli O157:H7. Elimination of E. coli O157:H7 during food processing and storage is a possible solution. Bacteriophages have a significant impact on bacterial populations in nature due to their ability to lyse their bacterial host. In the current study, a virulent bacteriophage, Ec_MI-02, was isolated from the feces of a wild pigeon in the United Arab Emirates (UAE) for potential future use as a bio-preservative or in phage therapy. Using a spot test and an efficiency of plating analysis, Ec_MI-02 was found to infect in addition to the propagation host, E. coli O157:H7 NCTC 12900, five different serotypes of E. coli O157:H7 (three clinical samples from infected patients, one from contaminated green salad, and one from contaminated ground beef). Based on morphology and genome analysis, Ec_MI-02 belongs to the genus Tequatrovirus under the order Caudovirales. The adsorption rate constant (K) of Ec_MI-02 was found to be 1.55 × 10-8 mL/min. The latent period was 50 min with a burst size of almost 10 plaque forming units (pfu)/host cell in the one-step growth curve when the phage Ec_MI-02 was cultivated using the propagation host E. coli O157:H7 NCTC 12900. Ec_MI-02 was found to be stable at a wide range of pH, temperature, and commonly used laboratory disinfectants. Its genome is 165,454 bp long with a GC content of 35.5% and encodes 266 protein coding genes. Ec_MI-02 has genes encoding for rI, rII, and rIII lysis inhibition proteins, which supports the observation of delayed lysis in the one-step growth curve. The current study provides additional evidence that wild birds could also be a good natural reservoir for bacteriophages that do not carry antibiotic resistance genes and could be good candidates for phage therapy. In addition, studying the genetic makeup of bacteriophages that infect human pathogens is crucial for ensuring their safe usage in the food industry.

A Versatile PdRu Bimetallic Nanoenzyme-Integrated Enzyme-Linked Immunosorbent Assay for Highly Sensitive Escherichia coli O157:H7 Detection.

Nanozymes have drawn much attention as an enzyme mimetic with low cost and stability in enhancing analytical performance. Herein, a peroxidase-mimicking nanozyme-improved enzyme-linked immunosorbent assay (ELISA) was developed employing the bimetallic PdRu nanozyme to replace the natural enzymes as a catalytic carrier for the sensing of Escherichia coli O157:H7 (E. coli O157:H7). The PdRu nanozyme displayed ultrahigh catalytic activity, possessing a catalytic rate that was 5-fold higher than horseradish peroxidase (HRP). In addition, PdRu exhibited great biological affinity with antibody (affinity constant was about 6.75 × 1012 M) and high stability. All those advantages ensure the successful establishment and the construction of a novel colorimetric biosensor for E. coli O157:H7 detection. PdRu-based ELISA not only achieved an ultrasensitive detection sensitivity (8.7 × 102 CFU/mL) by approximately 288-fold as compared to the traditional HRP-based ELISA and also possessed satisfactory specificity and reproducibility (relative standard deviation (RSD) < 10%). Furthermore, the feasibility of PdRu-ELISA was further evaluated by detecting E. coli O157:H7 in actual samples with satisfactory recoveries, indicating its potential for applications in bioassays and clinical diagnostics.

Antibacterial and antibiofilm performance of low-frequency ultrasound against Escherichia coli O157:H7 and its application in fresh produce

Ultrasound technology has been focused on due to its unique advantages in biofilm removal compared with traditional antibacterial methods. Herein, the anti-biofilm properties of low-frequency ultrasound (LFUS) were studied against Enterohemorrhagic Escherichia coli O157: H7 (E. coli O157:H7). After ultrasonication (20 kHz, 300 W) for 5 min, the removal rate of biofilm from polystyrene sheets reached up to 99.999 %. However, the bacterial cells could not be inactivated completely even extending the duration of ultrasonic irradiation to 30 min. Fortunately, this study indicated that LFUS could efficiently weaken the metabolic capacity and biofilm-forming ability of bacterial cells separated from biofilm. It could be associated with the removal of cell surface appendages and damage to cell membrane induced by mechanical vibration and acoustic cavitation. Besides, the genetic analysis proved that the transcription level of genes involved in curli formation was significantly down-regulated during ultrasonic irradiation, thus impeding the process of irreversible adhesion and cells aggregation. Finally, the actual application effect of LFUS was also evaluated in different fresh produces model. The results of this study would provide a theoretical basis for the further application of ultrasound in the food preservation.

Antimicrobial resistant Escherichia coli prevalence in freshwaters in Belgium and human exposure risk assessment

The objective of this work was to evaluate the antimicrobial resistant (AR) E. coli prevalence in recreational waters in Belgium and to assess the exposure risk for bathers. Nine stations were sampled during the 2021 bathing season. A total of 912 E. coli strains were isolated and tested by the disk diffusion method in accordance with EUCAST recommendations, including Extended-Spectrum Beta-Lactamase (ESBL) production. AR E. coli were counted at each bathing sites, 24% of strains were resistant to at least one antibiotic and 6% were Multi-Drug Resistant (MDR). A Multiple Antibiotic Resistance (MAR) index was calculated to compare the bathing sites. The Lesse river had the highest MAR index as well as the highest E. coli absolute abundance and the largest number of ESBL-producing E. coli. Conversely, the 3 lakes showed lower E. coli contamination levels and AR rates. A human health risk assessment of exposure to AR E. coli, based on the calculation of measured prevalence, was performed considering four different dose-response model scenarios. The human health risk (Pd) ranged from 10−9 to 0.183 (children). The exposure probabilities were low, except for scenario 3 (E. coli O157:H7), which is the most severe.

Nasal immunization with H7 flagellin protects mice against hemolytic uremic syndrome secondary to Escherichia coli O157:H7 gastrointestinal infection.

Shiga-toxin (Stx) producing Escherichia coli (STEC) O157:H7 is the most frequent serotype associated with hemolytic uremic syndrome (HUS) after gastrointestinal infections. Protection against HUS secondary to STEC infections has been experimentally assayed through the generation of different vaccine formulations. With focus on patients, the strategies have been mainly oriented to inhibit production of Stx or its neutralization. However, few approaches have been intended to block gastrointestinal phase of this disease, which is considered the first step in the pathogenic cascade of HUS. The aim of this work was to assay H7 flagellin as a mucosal vaccine candidate to prevent the systemic complications secondary to E. coli O157:H7 infections. The cellular and humoral immune response after H7 nasal immunization in mice were studied by the analysis of systemic and intestinal specific antibody production, as well as cytokine production and lymphocyte proliferation against H7 flagellin ex vivo. Immunized mice developed a strong and specific anti-H7 IgG and IgA response, at systemic and mucosal level, as well as a cellular Th1/Th2/Th17 response. H7 induced activation of bone marrow derived dendritic cells in vitro and a significant delayed-type hypersensitivity (DTH) response in immunized mice. Most relevant, immunized mice were completely protected against the challenge with an E. coli O157:H7 virulent strain in vivo, and surviving mice presented high titres of anti-H7 and Stx antibodies. These results suggest that immunization avoids HUS outcome and allows to elicit a specific immune response against other virulence factors.

Complete genome analysis of Tequatrovirus ufvareg1, a Tequatrovirus species inhibiting Escherichia coli O157:H7.

Bacteriophages infecting human pathogens have been considered potential biocontrol agents, and studying their genetic content is essential to their safe use in the food industry. Tequatrovirus ufvareg1 is a bacteriophage named UFV-AREG1, isolated from cowshed wastewater and previously tested for its ability to inhibit Escherichia coli O157:H7. T. ufvareg1 was previously isolated using E. coli O157:H7 (ATCC 43895) as a bacterial host. The same strain was used for bacteriophage propagation and the one-step growth curve. The genome of the T. ufvareg1 was sequenced using 305 Illumina HiSeq, and the genome comparison was calculated by VIRIDIC and VIPTree. Here, we characterize its genome and compare it to other Tequatrovirus. T. ufvareg1 virions have an icosahedral head (114 x 86 nm) and a contracted tail (117 x 23 nm), with a latent period of 25min, and an average burst size was 18 phage particles per infected E. coli cell. The genome of the bacteriophage T. ufvareg1 contains 268 coding DNA sequences (CDS) and ten tRNA genes distributed in both negative and positive strains. T. ufvareg1 genome also contains 40 promoters on its regulatory regions and two rho-independent terminators. T. ufvareg1 shares an average intergenomic similarity (VIRIDC) of 88.77% and an average genomic similarity score (VipTree) of 88.91% with eight four reference genomes for Tequatrovirus available in the NCBI RefSeq database. The pan-genomic analysis confirmed the high conservation of Tequatrovirus genomes. Among all CDS annotated in the T. ufvareg1 genome, there are 123 core genes, 38 softcore genes, 94 shell genes, and 13 cloud genes. None of 268 CDS was classified as being exclusive of T. ufvareg1. The results in this paper, combined with other previously published findings, indicate that T. ufvareg1 bacteriophage is a potential candidate for food protection against E. coli O157:H7 in foods.

Bacteriological Quality and Antimicrobial Resistant Patterns of Foodborne Pathogens Isolated from Commonly Vended Street Foods in Arba Minch Town, Southern Ethiopia.

Ready-to-eat foods sold by street vendors act as potential sources for the spread of various foodborne infectious diseases. Thus, local determination of the level of foodborne bacterial pathogens and their antimicrobial resistance pattern is essential. A community-based cross-sectional study was conducted from September 5th, 2022 to December 31th, 2022. The required data were collected by a structured questionnaire and observation checklist. Randomly selected street-vended foods were aseptically collected, and the bacteriological quality was assessed by using culture techniques. Different biochemical tests were used to identify and characterize bacterial isolates. The antimicrobial-resistant test for isolated foodborne bacterial pathogens was carried out by using the Kirby-Bauer disc diffusion method. The data were analyzed using SPSS version 22. A total of 34.2% (113/330) with a 95% CI of 29.1-39.4 of commonly consumed street-vended foods were identified as having unsatisfactory total mean aerobic bacterial counts (>105 CFU/g; 4.3 × 105 CFU/g). The mean total Enterobacteriaceae, coliform, and staphylococcal counts were found to be 1.4 × 105 CFU/g, 2.4 × 104 CFU/g, and 3.4 × 104 CFU/g, respectively. A total of 12.7% (42/330) of foodborne pathogens were recovered attributed to Staphylococcus aureus (31, 9.4%), Salmonella species (6, 1.8%), and E. coli O157:H7 (5, 1.5%). 6.5% and 16.1% of isolated S. aureus were found to be methicillin-resistant and multidrug-resistant (MDR), respectively. Additionally, 33.3% of Salmonella isolates and 40% of E. coli O157:H7 isolates were found to be MDR. Street-vended foods in this setting have a considerable number of unsatisfactory bacterial qualities, along with drug-resistant foodborne pathogens. Thus, more effective health education and training for vendors, regular inspections of vending sites, and regular surveillance of drug resistance patterns of foodborne pathogens are all essential.

Effect of high hydrostatic pressure treatment on the inactivation and sublethal injury of foodborne pathogens and quality of apple puree at different pH

The objectives of this study were to evaluate the survival of high hydrostatic pressure (HHP)-treated Salmonella Typhimurium, Escherichia coli O157:H7, and Listeria monocytogenes in apple puree, as well as to determine the levels of HHP-induced cell injury according to the pressure level, holding time, and pH of apple puree. Apple puree was inoculated with three foodborne pathogens and treated at pressures of 300–600 MPa for up to 7 min at 22 °C using HHP equipment. Increasing the pressure level and lowering the pH of apple puree led to larger microbial reductions, and E. coli O157:H7 showed higher resistance compared to S. Typhimurium and L. monocytogenes. Besides, approximately 5-log injured cells of E. coli O157:H7 were induced in apple puree at pH 3.5 and 3.8. HHP treatment at 500 MPa for 2 min effectively achieved complete inactivation of the three pathogens in apple puree at pH 3.5. For apple puree at pH 3.8, more than 2 min treatment of HHP at 600 MPa is seemingly needed to achieve complete inactivation of the three pathogens. Transmission electron microscopy analysis was conducted to identify ultrastructural changes in the injured or dead cells after HHP treatment. Plasmolysis and uneven cavities in the cytoplasm were observed in injured cells, and additional deformations, such as distorted and rough cell envelopes, and cell disruption occurred in dead cells. No changes in solid soluble content (SSC) and color of apple puree were observed after HHP treatment, and no differences were detected between control and HHP-treated samples during 10 d of storage at 5 °C. The results of this study could be useful in determining the acidity of apple purees or the treatment time at specific acidity levels when applying the HHP processing.