Antibiotic resistance profiling of pathogenic Enterobacteriaceae from Cluj-Napoca, Romania

Abstract

Introduction Members of the family Enterobacteriaceae are commonly identified in the clinical laboratory, being responsible for a substantial range of infections. This study aimed to investigate phenotypic and genotypic resistance traits in pathogenic Enterobacteriaceae isolated from outpatients in Cluj-Napoca, Romania. Methods Pathogenic Enterobacteriaceae were isolated from urinary tract infections, wound infections and persistent diarrhea in a private laboratory from Cluj-Napoca, Romania. Bacterial strains were biochemically identified and subjected to antimicrobial susceptibility testing by disk diffusion. The carriage of antibiotic resistance genes and of class 1 integron were assessed by PCR. Results E. coli and Enterobacter spp. were the most prevalent pathogens. High levels of resistance were observed against folate pathway inhibitors (74%), fluoroquinolones (49%) and penicillins (44%). The incidence of carbapenem resistance was 3%. The strains displaying phenotypic resistance were able to produce β-lactamase enzymes encoded by bla TEM , bla TEM-1 , bla SHV-1 and bla CTX-M , aminoglycoside modifying enzymes due to the carriage of aac(3)-IIIa, aac(6')-II and aac(6')-Ie-aph(2"), to possess fluoroquinolones resistance due to qnrS DNA gyrase protection proteins and resistance to folate pathway inhibitors due to dihydropteroate synthases encoded by sul1, sul2 and sul3 genes. The high frequency of intI1 integrase was associated to sulphonamide resistance (r=0.48; p<0.001) and also to fluoroquinolone resistance (r=0.27; p=0.011), but no significant associations in the co-occurrence of specific antibiotic resistance genes and intI1 were found in pathogenic Enterobacteriaceae. Conclusions An important proportion of pathogenic Enterobacteriaceae were multidrug resistant, due to a wide diversity of mechanisms encoding genetic resistance.

Revealing the CRISPR array in bacteria living in our organism

“…When looking at the isolates having a high number of spacers in CRISPR array we can hypothesize that this system is active. A previous study (Farkas et al, 2019) showed that these isolates bear a high number of ARGs encoding the resistance to different antibiotics. Assuming that CRISPR-Cas system inhibits the HGT of ARGs, most probably in these isolates, the CRISPR-Cas system is not active at present, ensuring the accumulation of ARGs.…”

“…Different bacteria have been isolated and identified in the hospital laboratory according to specific protocols, including Citrobacter, Enterobacter, Hafnia, Klebsiella, Proteus, Providencia, Salmonella, Yersinia and E. coli. (Farkas et al, 2019). Thus, 80 different isolates were analyzed for the presence of CRISPR arrays.…”

Prevalence of ESBL, AmpC and Carbapenemase-Producing Enterobacterales Isolated from Raw Vegetables Retailed in Romania

“…(n = 2) and E. coli, S. marcescens, M. morganii, K. oxytoca, P. vulgaris (each with n = 1). It is important to note that a recent study by Farkas et al (2019) concerning the antibiotic resistance profile of Enterobacteriaceae showed that the most prevalent pathogenic bacteria isolated from humans in the city of Cluj-Napoca, Romania, were E. coli and Enterobacter spp. [31].…”

Phenotypic and genotypic antibiotic susceptibility profiles of Gram-negative bacteria isolated from bloodstream infections at a referral hospital, Lusaka, Zambia

“…The primer selection and PCR protocol used was based on Farkas et. al [30]. PCR reactions were carried out using the following protocol: 2 μL of extracted DNA was combined with 12.5 μL of 2X master mix, 1 μL forward and 1 μL reverse primer, and the mixture was made up to a 25 μL volume with nuclease-free water.…”