Molecular study on virulence and resistance genes of ST131 clone (uropathogenic/enteropathogenic Escherichia coli) hybrids in children.

Aim: To analyze ST131 clones and other characteristics in uropathogenic and atypical enteropathogenic Escherichia coli hybrids. Methods: Samples were collected from children with urinary tract infections and underwent testing for antimicrobial susceptibility, multidrug resistance and extended-spectrum ß-lactamases, in vitro biofilm formation and virulence, resistance genes, hybrid pathotypes and ST131 clones. Results: E. coli isolates showed high levels of antibiotic resistance, extended-spectrum ß-lactamase production, virulence genes, multidrug resistance and biofilm formation. Four (5.0%) isolates were identified as uropathogenic/atypical enteropathogenic E. coli hybrids, all of which belonged to the high-risk ST131 clone. Conclusion: Our results provide promising insights about hybrid isolates and should be addressed to improve prevention measures for hybrid pathotypes.

Designing a novel multi-epitopes pan-vaccine against SARS-CoV-2 and seasonal influenza: in silico and immunoinformatics approach.

The merger of COVID-19 and seasonal influenza infections is considered a potentially serious threat to public health. These two viral agents can cause extensive and severe lower and upper respiratory tract infections with lung damage with host factors. Today, the development of vaccination has been shown to reduce the risk of hospitalization and mortality from the COVID-19 virus and influenza epidemics. Therefore, this study contributes to an immunoinformatics approach to producing a vaccine that can elicit strong and specific immune responses against COVID-19 and influenza A and B viruses. The NCBI, GISAID, and Uniprot databases were used to retrieve sequences. Linear B cell, Cytotoxic T lymphocyte, and Helper T lymphocyte epitopes were predicted using the online servers. Population coverage of MHC I epitopes worldwide for SARS-CoV-2, Influenza virus H3N2, H3N2, and Yamagata/Victoria were 99.93%, 68.67%, 68.38%, and 85.45%, respectively. Candidate epitopes were linked by GGGGS, GPGPG, and KK linkers. Different epitopes were permutated several times to form different peptides and then screened for antigenicity, allergenicity, and toxicity. The vaccine construct was analyzed for physicochemical properties, conformational B-cell epitopes, interaction with Toll-like receptors, and IFN-gamma-induced. Immune stimulation response of final construct was evaluated using C-IMMSIM. Eventually, the final construct sequence was codon-optimized for Escherichia coli K12 and Homo sapiens to design a multi-epitope vaccine and mRNA vaccine. In conclusion, due to the variable nature of SARS-CoV-2 and influenza proteins, the design of a multi-epitope vaccine can protect against all their standard variants, but laboratory validation is required.Communicated by Ramaswamy H. Sarma.

Application and development of fecal microbiota transplantation in the treatment of gastrointestinal and metabolic diseases: A review.

Fecal microbiota transplantation (FMT) restores a balanced intestinal flora, which helps to cure recurrent Clostridium difficile infections (RCDI). FMT has also been used to treat other gastrointestinal diseases, including inflammatory bowel disease (IBD), irritable bowel syndrome (IBS), and chronic constipation, as well as a variety of non-GI disorders. The purpose of this review is to discuss gut microbiota and FMT treatment of GI and non-GI diseases. An imbalanced gut microbiota is known to predispose one to Clostridium difficile infections (CDI), IBD, and IBS. However, the complex role of the gut microbiota in maintaining health is a newer concept that is being increasingly studied. The microbiome plays a major role in cellular immunity and metabolism and has been implicated in the pathogenesis of non-GI autoimmune diseases, chronic fatigue syndrome, obesity, and even some neuropsychiatric disorders. Many recent studies have reported that viral gastroenteritis can affect intestinal epithelial cells, and SARS-CoV-2 virus has been identified in the stool of infected patients. FMT is a highly effective cure for RCDI, but a better understanding of the gut microbiota in maintaining health and controlled studies of FMT in a variety of conditions are needed before FMT can be accepted and used clinically.

Rapid Detection of vanA Resistance Gene from E. faecalis Clinical Isolates Using Duplex Loop-Mediated Isothermal Amplification and Triplex PCR Assay.

Today, the spread of vancomycin-resistant strains isolated from Enterococcus faecalis (E. faecalis) has become a major health concern worldwide. Therefore, it is essential to provide a rapid and sensitive assay for identifying vanA gene for timely and appropriate antimicrobial control of resistant enterococcal infections. For this purpose, a cross-sectional study was performed on different clinical specimens of enterococci from Imam Reza hospital, Kermanshah, Iran. The antimicrobial susceptibility testing was determined by disk diffusion and MIC methods. Triplex-PCR and duplex-LAMP assays were also used to identify vanA E. faecalis resistance gene isolates. The results of this study shown that out of 108 Enterococcus isolates, 86, 18, 2, 1, and one isolates of E. faecalis, E. faecium, E. avium, E. psudoavium, and E. raffinosus were identified, respectively. On the other hand, E. faecalis was confirmed in 87 and 88 isolates using duplex-LAMP and triplex PCR, respectively. The LAMP primer set designed in this study can reliably identify seven distinct regions of the vanA gene, and finally the sensitivity, specificity, and the positive and negative predictive values of LAMP assay were shown to be 94.19%, 72.73%, 76.19%, and 93.10%, respectively. In general, sample processing, isothermal reaction and result reporting were completed using the LAMP assay in 75 minutes. Our findings suggest that LAMP assay has been approved as an alternative to the vancomycin resistance Enterococcus genotype (vanA and vanB) compared to other methods and has the advantage of being rapid, time-consuming, and easy for diagnosis.

An update review of globally reported SARS-CoV-2 vaccines in preclinical and clinical stages.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of the rapidly spreading pandemic COVID-19 in the world. As an effective therapeutic strategy is not introduced yet and the rapid genetic variations in the virus, there is an emerging necessity to design, evaluate and apply effective new vaccines. An acceptable vaccine must elicit both humoral and cellular immune responses, must have the least side effects and the storage and transport systems should be available and affordable for all countries. These vaccines can be classified into different types: inactivated vaccines, live-attenuated virus vaccines, subunit vaccines, virus-like particles (VLPs), nucleic acid-based vaccines (DNA and RNA) and recombinant vector-based vaccines (replicating and non-replicating viral vector). According to the latest update of the WHO report on April 2nd, 2021, at least 85 vaccine candidates were being studied in clinical trial phases and 184 candidate vaccines were being evaluated in pre-clinical stages. In addition, studies have shown that other vaccines, including the Bacillus Calmette-Guérin (BCG) vaccine and the Plant-derived vaccine, may play a role in controlling pandemic COVID-19. Herein, we reviewed the different types of COVID-19 candidate vaccines that are currently being evaluated in preclinical and clinical trial phases along with advantages, disadvantages or adverse reactions, if any.

The Prevalence of Shiga Toxin-Producing Escherichia coli and Enteropathogenic Escherichia coli Isolated from Raw Chicken Meat Samples.

BACKGROUND: Shiga toxin-producing Escherichia coli (STEC) is known as a crucial zoonotic food-borne pathogen. A total of 257 raw chicken meat samples were collected from different markets in Hamadan, west of Iran, from January 2016 to May 2017. MATERIALS AND METHODS: The samples were cultured in selective and differential culture media, and the virulence genes of E. coli isolates were analyzed by PCR assay. The antibiotic resistance patterns of E. coli isolates were determined by the disk diffusion method. The genetic relatedness of the E. coli O157 isolates was analyzed by ERIC-PCR. RESULTS: In total, 93 (36% ± 3.12) of the isolates were identified as E. coli in this study. Based on serological and microbiological tests, 36 (38.7% ± 9.9), 7 (7.5% ± 5.35), and 12 (12.9% ± 6.81) of the E. coli isolates were characterized as STEC, enteropathogenic E. coli (EPEC), and attaching and effacing E. coli (AEEC) strains, respectively. A high level of resistance to nalidixic acid (91.4% ± 5.7), tetracycline (89.2% ± 6.31), ampicillin (82.8% ± 7.67), and trimotoprime-sulfametoxazole (71% ± 9.22) was detected among the E. coli isolates. The analysis of the ERIC-PCR results showed five different ERIC types among the E. coli O157 isolates. CONCLUSIONS: Based on our findings, control and check-up of poultry meats should be considered as a crucial issue for public health.

Distribution of Iron Uptake Systems Encoding Genes Among the Clinical Isolates of Escherichia coli Compared to Foodstuffs Isolates.

INTRODUCTION: Bacteria require iron ions to grow and infect the host, which, by using iron uptake systems, acquire free iron from their host cell. Escherichia coli is one of the most important pathogens to cause food poisoning and clinical infections. The aim of this study was to assess the distribution of iron uptake systems encoding genes in clinical isolates of E.coli compared to food samples isolates. MATERIALS AND METHODS: This investigation was conducted to determine the prevalence of E. coli isolated from various sources of food and clinical specimens. The E. coli isolates confirmed by the standard microbiological methods. The isolates were examined for the presence of iut A and iuc A genes by specific primers using the polymerase chain reaction technique. RESULTS: A total of 100 and 50 isolates of E. coli were collected from clinical samples and foodstuffs, respectively. The prevalence of E. coli in the food and clinical samples was 33.33% and 64.10%, respectively. The frequency of iut A and iuc A genes in the food and clinical isolates were 76%-84% and 86% - 83%, respectively. CONCLUSION: Our results showed that the prevalence of E. coli isolates with iut A and iuc A genes was relatively higher compared to many previous studies. The existence of these genes in E. coli strains is likely to be related to pathogenicity in those strains, which requires further studies in the future.

Identification of Quinolone and Colistin Resistance Genes in Escherichia Coli Strains Isolated from Mucosal Samples of Patients with Colorectal Cancer and Healthy Subjects.

INTRODUCTION: Antibiotic resistance and extensive use of antibiotics are amongst the major causes of failure in antibiotic treatment. The purpose of this study was to investigate antibiotic resistance patterns and to identify resistance genes of quinolones and colistin in Escherichia coli. There are a very few patents on E. coli isolated from colorectal cancer. So, this study demonstrates that some bacteria resistant to ciprofloxacin have not resistance genes.Moreover, new patterns for E. coli are presented for isolates of patients with colorectal cancer. MATERIALS AND METHODS: Of the three healthy people, inflammatory bowel diseases (IBD) patients and colorectal cancer patients, 40 E. coli strains isolated after confirmation by biochemical and molecular methods. The susceptibility of isolates to antibiotics was investigated using disk diffusion test. After deoxyribonucleic acid (DNA) extraction, polymerase chain reaction (PCR) was used to identify genes encoding resistance to ciprofloxacin (qnr A, qnr B) and colistin (mcr-1). RESULTS: The results showed that E. coli isolates from colorectal cancer patients had the highest resistance to piperacillin (67.5%), ceftazidime (47.5%), and cefepime (42.5%). Also, E. coli strains isolated from IBD patients showed resistance to antibiotic ceftazidime 13%. More than 95% of E. coli strains isolated from healthy people were susceptible to antibiotics. Based on the results, 18 (15%) E. coli strains showed resistance to ciprofloxacin. The qnr A gene was detected in 61.11% isolates; however, qnr B was detected in 9 (50%) isolates. Isolates resistant to colistin were not observed. CONCLUSION: These findings indicate increased resistance of E. coli to ciprofloxacin in comparison with prior studies. Further research in this field will increase our knowledge and more effective exposure to the antibiotic resistance of the pathogenic microorganisms.

Molecular analysis of Pseudomonas aeruginosa isolated from clinical, environmental and cockroach sources by ERIC-PCR.

OBJECTIVE: The objective of this study was to investigate the antibiotic susceptibility, virulence factors and clonal relationship among Pseudomonas aeruginosa isolated from environmental sources, hospitalized patients and the surfaces of cockroaches in the ICUs of four hospitals in Hamadan, west of Iran. A total of 237, 286 and 156 bacterial isolates were collected from clinical, environmental and cockroach sources respectively from May to September, 2017. The antimicrobial susceptibility was determined using disk diffusion method. The virulence factors, exotoxins A, S and U were detected by PCR. The genetic linkage of P. aeruginosa isolates were analyzed by Enterobacterial Repetitive Intergenic Consensus (ERIC)-PCR. RESULTS: According to our findings, 58 (24.4%), 46 (16%) and 5 (3.25) P. aeruginosa were isolated from clinical, environmental and cockroach samples respectively. The MDR phenotypes were detected in 18 (45%) and 15 (37.5%) of clinical and environmental strains. The environmental isolates harbored more exoA and exoS than did clinical isolates. Genetic diversity was established among P. aeruginosa isolates as 14 different ERIC fingerprints were detected. The clonal relationships was detected among clinical, environmental and cockroach isolates. Our results highlighted the importance of identifying and controlling the potential sources of P. aeruginosa infections in hospitals.