Molecular Identification of Extended-Spectrum ß-lactamase and Integron Genes in Klebsiella Pneumonia.

INTRODUCTION: Infections caused by Gram negative bacteria, producing extended-spectrum ß-lactamase, including Klebsiella pneumoniae are increasing all over the world with high morbidity and mortality. The aim of the present study was determined antimicrobial profile susceptibility and the prevalence of antibiotic resistance genes by multiplex PCR. METHODS: In the present study, we obtained one-hundred isolates of K. pneumoniae from different clinical samples. The antibiotic susceptibility testing was done in thirteen antibiotic and, therefore, M-PCRs were conducted using the DNA amplification for detection of ESBLs (blaTEM, blaCTX-M, blaSHV) and int (I, II, III) genes. RESULTS: The results of resistance to amoxicillin/clavulanate, ciprofloxacin, amikacin, trimethoprim-sulfamethoxazole, cefotaxime, ampicillin, aztreonam, imipenem, gentamicin, ceftazidime, Cefepime, ceftriaxone and levofloxacin were obtained 37%, 37%, 93%, 84%, 52%, 87%, 59%, 8%, 24%, 67%, 52%, 43% and 26%, respectively. The frequency of the extended-spectrum ß-lactamase K. pneumoniae was obtained 37%. The prevalence of resistance genes of ESBLs in the M-PCR method showed that the blaTEM, blaCTX and blaSHV were 38%, 24% and 19%, respectively, however, only 8 (8%) out of 100 isolates were found to have positive outcomes for the existence of class 1 integrons and there were no detected class 2 or class 3 integrons. CONCLUSIONS: Our results recommend the likely co-carriage of some ESBLs genes and antibiotic resistance integrons on the same plasmids harboring the MDR genes.

Antifungal activity and influence of propolis against germ tube formation as a critical virulence attribute by clinical isolates of Candida albicans.

OBJECTIVE: The potentiality to switch between yeast and hyphal morphologies plays an important role in the virulence of Candida albicans (C. albicans). The objective of the present study was to evaluate the chemical composition, antifungal activity and influence of propolis upon germ tube formation of C. albicans clinical isolates. METHODS: The major compounds of propolis ethanolic extract (PEE) was revealed by gas chromatography-mass spectrometry (GC/MS) analysis. Broth microdilution method was used for determination of minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) of this extract. C. albicans isolates were exposed to sub-MIC concentrations of propolis and germ tube reduction percent (GRP) and morphological transition from yeast to filamentous form were monitored microscopically. RESULTS: Phenolic compounds, aromatic acids, aliphatic acids, sugars and polycyclic aromatic hydrocarbons were the major compound classes in PEE. Mean of MIC and MFC values of PEE against C. albicans isolates were 360.6µg·mL-1 and 1250.1µg·mL-1, respectively. The increase in concentration led to a significant reduction in germ tube formation, so mean of GRP was 36.7±11.1 and 22±10.1 at concentration of ½ MIC and » MIC of PEE, respectively. CONCLUSIONS: In conclusion, the present study indicated that PEE has interesting potential as a therapeutic agent that targets germ tube formation by C. albicans as a critical virulence factor.

Molecular characterization of Salmonella enterica serotype Enteritidis isolates from food and human samples by serotyping, antimicrobial resistance, plasmid profiling, (GTG)5-PCR and ERIC-PCR.

In recent years, Salmonella enterica serovar Enteritidis has been a primary cause of human salmonellosis in many countries. The major objective of this study was to investigate genetic diversity among Salmonella Enteritidis strains from different origins (food and human) by Enterobacterial Repetitive Intergenic Consensus (ERIC) -PCR, as well as to assess their plasmid profiling and antimicrobial resistance. A total of 30 Salmonella Enteritidis isolates, 15 from food samples (chicken, lamb, beef and duck meats) and 15 from clinical samples were collected in Tehran. Identification of isolates as Salmonella was confirmed by using conventional standard biochemical and serological tests. Multiplex-PCR was used for serotyping of isolates to identify Salmonella Enteritidis. Antimicrobial susceptibility testing to 16 agents founds drug resistance patterns among Salmonella Enteritidis isolates. No resistance was observed to cephalexin, ceftriaxone, ceftazidime and cefotaxime, ciprofloxacin, imipenem or meropenem, chloramphenicol and gentamicin. The highest resistance (96.7%) was observed to nitrofurantoin. Seven plasmid profiles (P1-P7) were detected, and a 68-kb plasmid was found in all isolates. Two different primers; ERIC and (GTG)5 were used for genotyping, which each produced four profiles. The majority of clinical and food isolates fell into two separate common types (CTs) with a similar percentage of 95% by ERIC-PCR. Using primer (GTG)5, 29 isolates incorporated in three CTs with 70% of isolates showing a single banding pattern. Limited genetic diversity among human and food isolates of Salmonella Enteritidis may indicate that contaminated foods were possibly the source of human salmonellosis. These results confirmed that ERIC-PCR genotyping has limited discriminatory power for Salmonella Enteritidis of different origin.