Isolation, characterization, molecular analysis and application of bacteriophage DW-EC to control Enterotoxigenic Escherichia coli on various foods

Among food preservation methods, bacteriophage treatment can be a viable alternative method to overcome the drawbacks of traditional approaches. Bacteriophages are naturally occurring viruses that are highly specific to their hosts and have the capability to lyse bacterial cells, making them useful as biopreservation agents. This study aims to characterize and determine the application of bacteriophage isolated from Indonesian traditional Ready-to-Eat (RTE) food to control Enterotoxigenic Escherichia coli (ETEC) population in various foods. Phage DW-EC isolated from Indonesian traditional RTE food called dawet with ETEC as its host showed a positive result by the formation of plaques (clear zone) in the bacterial host lawn. Transmission electron microscopy (TEM) results also showed that DW-EC can be suspected to belong to the Myoviridae family. Molecular characterization and bioinformatic analysis showed that DW-EC exhibited characteristics as promising biocontrol agents in food samples. Genes related to the lytic cycle, such as lysozyme and tail fiber assembly protein, were annotated. There were also no signs of lysogenic genes among the annotation results. The resulting PHACTS data also indicated that DW-EC was leaning toward being exclusively lytic. DW-EC significantly reduced the ETEC population (P ≤ 0.05) in various food samples after two different incubation times (1 day and 6 days) in chicken meat (80.93%; 87.29%), fish meat (63.78%; 87.89%), cucumber (61.42%; 71.88%), tomato (56.24%; 74.51%), and lettuce (46.88%; 43.38%).

Evaluation of microbial contamination in cold dishes and Prevalence of food-borne pathogens in the Jilin Province

In this study, we evaluated the microbial contamination status of cold dishes consumed by residents of Jilin Province and investigated to determine the incidence of four pathogenic bacteria in cold dishes. A total of 300 samples of cold dishes including meat, vegetable and mixed products, were collected from three different purchasing places: supermarkets, farmers' markets and mobile vendors. Live bacteria were isolated using conventional culture methods. After separation, a quick and easy polymerase chain reaction (PCR) was used to detect Listeria monocytogenes , Staphylococcus aureus , Enterotoxic Escherichia coli and Salmonella . The results showed that the total number of microbial colonies in the vegetable samples exceeded the standard rate of 8%, and the total number of microbial colonies in the meat and mixed samples did not exceed the standard. The total microbial colony count exceeded the standard in all three different procurement sites, with the highest exceedance of 7.4% in the mobile vendor sites. The detection rates of Enterotoxigenic Escherichia coli , Staphylococcus aureus , L. monocytogenes and Salmonella among the four pathogenic bacteria detected in all samples were 4.3%, 3.3%; 3.0%; and 1.0%, respectively. This study can be used to qualitatively assess the microbiological quality associated with cold dishes. It provides data to support the detection of possible food safety problems.

Oral Immunogenicity of Enterotoxigenic Escherichia coli Outer Membrane Vesicles Encapsulated into Zein Nanoparticles Coated with a Gantrez® AN–Mannosamine Polymer Conjugate

Enterotoxigenic Escherichia coli (ETEC) represents a major cause of morbidity and mortality in the human population. In particular, ETEC infections affect children under the age of five from low-middle income countries. However, there is no licensed vaccine against this pathogen. ETEC vaccine development is challenging since this pathotype expresses a wide variety of antigenically diverse virulence factors whose genes can be modified due to ETEC genetic plasticity. To overcome this challenge, we propose the use of outer membrane vesicles (OMVs) isolated from two ETEC clinical strains. In these OMVs, proteomic studies revealed the presence of important immunogens, such as heat-labile toxin, colonization factors, adhesins and mucinases. Furthermore, these vesicles proved to be immunogenic after subcutaneous administration in BALB/c mice. Since ETEC is an enteropathogen, it is necessary to induce both systemic and mucosal immunity. For this purpose, the vesicles, free or encapsulated in zein nanoparticles coated with a Gantrez®–mannosamine conjugate, were administered orally. Biodistribution studies showed that the encapsulation of OMVs delayed the transit through the gut. These results were confirmed by in vivo study, in which OMV encapsulation resulted in higher levels of specific antibodies IgG2a. Further studies are needed to evaluate the protection efficacy of this vaccine approach.

Quality of tomato (Solanum lycopersicum L.) fruits inoculated with Escherichia coli under different storage conditions

The study evaluated the effect of storage temperatures of 7 and 22 °C for 168 h on tomatoes (Charleston cv.) inoculated with 107 CFU mL-1 of the enterotoxigenic Escherichia coli pathogroup (ETEC) strain on color indexes (hue angle, h°, and chroma, C*), firmness, titratable acidity (% citric acid), ascorbic acid, total soluble sugars and reducing sugars (glucose, fructose, and sucrose). ETEC survived with populations of 7 and 9.2 Log CFU g-1 at 7 and 22 °C, respectively until 120 h. Bacterial adherence and colonization under both storage conditions were confirmed by scanning electron microscopy. The index C* and ascorbic acid had higher values at 22 °C, while the parameters h°, firmness, and citric acid had lower values at the same storage temperature. At 7 °C, the concentration of total soluble sugars was affected; glucose and fructose showed lower values (0.054 and 0.057 g 100 g-1, respectively). Finally, the inoculated fruits exhibited significant differences in the parameters of consumer preference of fresh tomatoes such as color, firmness, sugars, and organic acids, which were affected depending on the storage temperature.

Evaluation of the Antibacterial and Prebiotic Potential of Ascophyllum nodosum and Its Extracts Using Selected Bacterial Members of the Pig Gastrointestinal Microbiota

Ascophyllum nodosum and its extracts are promising antibacterial and prebiotic dietary supplements for pigs. The objectives of this study were to evaluate the effects of the increasing concentrations of: (1) two whole biomass samples of A. nodosum with different harvest seasons, February (ANWB-F) and November (ANWB-N), in a weaned pig faecal batch fermentation assay, and (2) A. nodosum extracts produced using four different extraction conditions of a hydrothermal-assisted extraction methodology (ANE1–4) and conventional extraction methods with water (ANWE) and ethanol (ANEE) as solvent in individual pure culture growth assays using a panel of beneficial and pathogenic bacterial strains. In the batch fermentation assay, ANWB-F reduced Bifidobacterium spp. counts (p < 0.05) while ANWB-N increased total bacterial counts and reduced Bifidobacterium spp. and Enterobacteriaceae counts (p < 0.05). Of the ANE1–4, produced from ANWB-F, ANWE and ANEE that were evaluated in the pure culture growth assays, the most interesting extracts were the ANE1 that reduced Salmonella Typhimurium, enterotoxigenic Escherichia coli and B. thermophilum counts and the ANE4 that stimulated B. thermophilum growth (p < 0.05). In conclusion, the extraction method and conditions influenced the bioactivities of the A. nodosum extracts with ANE1 and ANE4 exhibiting distinct antibacterial and prebiotic properties in vitro, respectively, that merit further exploration.

Sự hiện diện của một số gene beta-lactamase và sự đề kháng kháng sinh của vi khuẩn Enterohemorrhagic Escherichia coli và Enterotoxigenic Escherichia coli phân lập từ bò tại tỉnh Bến Tre

Nghiên cứu được thực hiện nhằm khảo sát sự đề kháng kháng sinh và tỷ lệ hiện diện gene mã hoá beta-lactamase trên 21 chủng Enterohemorrhagic Escherichia coli (EHEC) và 38 chủng Enterotoxigenic Escherichia coli (ETEC) phân lập từ bò tại tỉnh Bến Tre. Các chủng EHEC có tỷ lệ đề kháng cao với colistin (71,43%) và ampicillin (61,90%), nhưng còn tỷ lệ nhạy cảm rất cao với doxycycline (100,00%), amikacin (95,24%). Trong 14 kiểu hình đa kháng của các chủng EHEC, phổ biến là kiểu hình Cz+Co (9,52%). Đối với ETEC, các chủng này còn nhạy cảm cao với các loại kháng sinh, nhạy cảm 100% đối với gentamicin, amikacin, levofloxacin và ofloxacin. Trong 13 kiểu hình đa kháng của các chủng ETEC, kiểu hình Am+Ac+Sm và Am+Cu+Co xuất hiện phổ biến (5,26%). Khảo sát bằng phương pháp PCR cho thấy trên các chủng EHEC và ETEC có sự hiện diện của 4/5 gene beta-lactamase được khảo sát. Gene blaampC chiếm tỷ lệ cao nhất trên EHEC, ETEC với tỷ lệ lần lượt là 57,14%, 42,11%. Không có sự hiện diện của gene blaCMY trên cả hai chủng. Có sự hình thành 3 kiểu hình kết hợp gene beta-lactamse, trong đó kiểu hình blaampC+blaTEM được ghi nhận nhiều nhất (10,17%).