In the present study a total of 200 Klebsiella pneumoniae isolates were collected from patients with urinary tract infections (UTIs) in Tehran, Iran. Antibiotic resistance was determined by disk diffusion and broth dilution methods. Detection of extended-spectrum β-lactamases (ESBLs) and AmpCs was performed using phenotypic tests. Polymerase chain reaction (PCR) was applied to detect the ESBL, AmpC, and integron genes. Analysis of AmpC and cassette arrays of integron genes was performed using DNA sequencing. Plasmids were analyzed by PCR-based replicon typing and conjugation. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) were applied to explore the genomic relatedness among the isolates. The highest levels of resistance were observed against ampicillin (100%), followed by piperacillin (57.5%), ceftazidime (46%), trimethoprim/sulfamethoxazole (44%), ciprofloxacin (32.5%), and imipenem (19%). Approximately, 66.5% of isolates harbored at least one of the beta-lactamase genes (bla
TEM, bla
SHV, bla
CTX-M, and bla
OXA-1). In addition, 22.5% of isolates carried at least one of the AmpC genes including bla
DHA and bla
CIT. Integron class I was the most prevalent integron among resistant isolates. According to the results of replicon typing, IncFII, IncL/M, and IncA/C were the most frequent replicons, respectively. All selected isolates were able to transfer bla
CTX-M, also two isolates transferred the bla
DHA-1 gene to Escherichia coli K12 through conjugation. Finally, 21 isolates were categorized into 4 pulsotypes and 11 unique clusters in PFGE. MLST identified ST147 and ST11 sequence types but ST147 was the most prevalent in the current study.
The rate of nasal carriage is higher in patients with MS. Our study's results suggest that further investigation into whether there is a connection between MS and nasal exposure to staphylococcal superantigens is warranted.
Backgrounds: Bacteriophage therapy could be an alternative strategy for the treatment of antibiotic-resistant bacteria. This study aimed to evaluate the antibacterial activities of isolated bacteriophages against methicillin-resistant Staphylococcus aureus (MRSA) isolates. Materials & Methods: A total of 16 clinical isolates of MRSA were collected from medical diagnostic laboratories in Tehran, Iran. A specific bacteriophage was isolated from hospital sewage using double-layer agar. Phage morphology was evaluated by transmission electron microscopy (TEM). Different bacteria were selected to determine the bacteriophage host range using spot test. Phage susceptibility to temperature and pH was evaluated by doublelayer agar method. In vitro assay was carried out on human epithelial type 2 (HEp-2) cells to investigate the effect of bacteriophage on the adhesion of MRSA to human epithelial cells. Findings: TEM suggested the Myoviridae family for the isolated phage. The effective titer of bacteriophages was 1.8×10 7 PFU/mL. The isolated bacteriophage was stable at 4 ˚C and pH=8. The isolated bacteriophage was specific for all clinical isolates of MRSA and had no lytic activity against other pathogenic bacteria. In evaluating the binding and invasion of MRSA to the HEp-2 cell line, as expected, the lytic activity of specific bacteriophages was observed following inoculation.
Conclusion:The specificity and lytic activity of this phage on MRSA and MRSA-infected HEp-2 cell line emphasized that the isolated bacteriophage may serve as an effective prophylactic and alternative therapeutic agent in hospital settings.
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