Uropathogenic E.coli (UPEC) are among major pathogens causing urinary tract infections. Virulence factors are mainly responsible for the severity of these emerging infections. This study was planned to investigate the distribution of virulence genes and cytotoxic effects of UPEC isolates with reference to phylogenetic groups (B2, B1, D and A) to understand the presence and impact of virulence factors in the severity of infection in Faisalabad region of Pakistan.

The zoonotic serovars of Salmonella enterica particularly Typhimurium and Enteritidis pose a continuous global threat to poultry industry and public health. We report the prevalence of Salmonella Typhimurium and Salmonella Enteritidis serovars in local poultry, phenotypic antimicrobial resistance profiling, and molecular detection of antimicrobial resistance and virulence genes. A total of 340 clinical samples were collected and 239 carried Salmonella , which were identified by genus-specific PCR ( invA gene) and by matrix-assisted laser desorption/ionization time-of-flight. The 68 and 22 isolates were confirmed as Salmonella Typhimurium ( stm gene) and Salmonella Enteritidis ( sdfI gene) respectively. Pulsed-field gel electrophoresis (PFGE) revealed 27 and 9 PFGE types of Salmonella Typhimurium and Salmonella Enteritidis respectively. Among 24 antimicrobials tested, highest resistance was observed against pefloxacin while highest susceptibility was found for ertapenem in Salmonella Typhimurium and aztreonam in Salmonella Enteritidis. All isolates were found multiple drug resistant, 98.8% as motile and 8.8% as extended spectrum beta lactamase producers. Most prevalent resistance gene in Salmonella Typhimurium was parE (69.1%) while in Salmonella Enteritidis blaTEM-1 (72.7%). High prevalence of SopE gene in Salmonella Typhimurium (91.1%) and Salmonella Enteritidis (81.8%) indicated their zoonotic potential. The study is first of its kind from this region and highlights the emerging trends of antimicrobial resistance of global concern.

Avian pathogenic Escherichia coli (APEC) are major bacterial pathogens of commercial poultry contributing to extensive economic losses and contamination of the food chain. One of the initial steps in bacterial infection and successful colonization of the host is adhesion to the host cells. A random transposon mutant library (n=1300) of APEC IMT 5155 was screened phenotypically for adhesion to chicken (CHIC-8E11) and human (LoVo) intestinal epithelial cell lines. The detection and quantification of adherent bacteria was performed by a modified APEC-specific antibody staining assay using fluorescence microscopy coupled to automated VideoScan Technology. Eleven mutants were found to have significantly altered adhesion to the cell lines examined. Mutated genes in these 11 ‘adhesion-altered mutants’ were identified by arbitrary PCR and DNA sequencing. The genes were amplified from wildtype APEC IMT 5155, cloned and transformed into the respective ‘adhesion-altered mutants’ and complementation was determined in adhesion assays. Here, we report contributions of the fdtA, rluD, yjhB, ecpR and fdeC genes of APEC in adhesion to chicken and intestinal cell lines. Identification of the roles of these genes in APEC pathogenesis will contribute in prevention and control of APEC infections. IMPORTANCE Avian pathogenic E. coli are not only pathogenic bacteria for commercial poultry but can also cause foodborne infections in humans utilizing the same attachment and virulence mechanisms. Our aim was to identify genes of avian pathogenic E. coli involved in adhesion to chicken and human cells in order to understand the colonization and pathogenesis of these bacteria. In contrast to the recent studies based on genotypic and bioinformatics data, we have used a combination of phenotypic and genotypic approaches for identification of novel genes contributing to adhesion in chicken and human cell lines. Identification of adhesion factors remains important, as antibodies elicited against such factors have shown potential to block colonization and ultimately prevent disease as prophylactic vaccines. Therefore, the data will augment the understanding of disease pathogenesis and ultimately in designing strategies against the infections.