Characteristics of Quinolone Resistance in Escherichia coli Isolates from Humans, Animals, and the Environment in the Czech Republic

Abstract

Escherichia coli is a common commensal bacterial species of humans and animals that may become a troublesome pathogen causing serious diseases. The aim of this study was to characterize the quinolone resistance phenotypes and genotypes in E. coli isolates of different origin from one area of the Czech Republic. E. coli isolates were obtained from hospitalized patients and outpatients, chicken farms, retailed turkeys, rooks wintering in the area, and wastewaters. Susceptibility of the isolates grown on the MacConkey agar with ciprofloxacin (0.05 mg/L) to 23 antimicrobial agents was determined. The presence of plasmid-mediated quinolone resistance (PMQR) and ESBL genes was tested by PCR and sequencing. Specific mutations in gyrA, gyrB, parC, and parE were also examined. Multilocus sequence typing and pulsed-field gel electrophoresis were performed to assess the clonal relationship. In total, 1050 E. coli isolates were obtained, including 303 isolates from humans, 156 from chickens, 105 from turkeys, 114 from the rooks, and 372 from wastewater samples. PMQR genes were detected in 262 (25%) isolates. The highest occurrence was observed in isolates from retailed turkey (49% of the isolates were positive) and inpatients (32%). The qnrS1 gene was the most common PMQR determinant identified in 146 (56%) followed by aac(6′)-Ib-cr in 77 (29%), qnrB19 in 41 (16%), and qnrB1 in 9 (3%) isolates. All isolates with high level of ciprofloxacin resistance (>32 mg/L) carried double or triple mutations in gyrA combined with single or double mutations in parC. The most frequently identified substitutions were Ser(83)Leu; Asp(87)Asn in GyrA, together with Ser(80)Ile, or Glu(84)Val in ParC. Majority of these isolates showed resistance to beta-lactams and multiresistance phenotype was found in 95% isolates. Forty-eight different sequence types among 144 isolates analyzed were found, including five major clones ST131 (26), ST355 (19), ST48 (13), ST95 (10), and ST10 (5). No isolates sharing 100% relatedness and originating from different areas were identified. In conclusion, our study identified PMQR genes in E. coli isolates in all areas studied, including highly virulent multiresistant clones such as ST131 producing CTX-M-15 beta-lactamases.

Molecular Epidemiology of Dairy Cattle-Associated Escherichia coli Carrying bla CTX-M Genes in Washington State

“…ST744 has been frequently reported from widely dispersed locations around the world, including from food animals in Australia (20), broiler chickens in Algeria (21), bovine mastitis in Germany (22), cattle in France 23, and hospitalized patients in Hong Kong (24), many with ESBL and/or carbapenem resistance characteristics. ST48 has also frequently been reported from diverse sources and locations and is reportedly associated with resistance (25)(26)(27)(28)(29)(30). The IncI1 and IncF plasmid families were the most frequent replicon types detected in this study, which is consistent with reports that IncI1 and IncF were the most frequent replicon types among bla CTX-M-15 -carrying plasmids in Enterobacteriaceae from human, livestock, and environmental sources in European studies (31)(32)(33).…”

High Genomic Diversity and Heterogenous Origins of Pathogenic and Antibiotic-Resistant Escherichia coli in Household Settings Represent a Challenge to Reducing Transmission in Low-Income Settings

“…The plasmid-mediated quinolone resistance (PMQR) genes QnrS1 and QnrB4 were detected in eight E. coli; however, no clinical resistance to ciprofloxacin, based on CLSI breakpoints, was observed in these isolates, except for one soil isolate that coharbored both genes. QnrS1 and QnrB4 are known to provide a low level of resistance, while mutations in the genes encoding DNA gyrase and topoisomerase IV are associated with observable resistance to ciprofloxacin and/or nalidixic acid (31), as in the case of seven E. coli isolates of this study (Table 1). Resistance to azithromycin (macrolide), detected only in E. coli from human and soil origin, was observed in the five isolates where the macrolide-associated gene(s) mph(A) and/or ermB was detected ( Table 1).…”

Occurrence of bacteria with a dangerous extent of antibiotic resistance in poultry in the Central Region of Moravia

“…To determine the genetic relatedness of E. coli isolates, macrorestriction analysis of chromosomal DNA was performed using pulsed-field gel electrophoresis (PFGE). Genomic DNAs prepared in accordance with a previously described procedure (Husickova et al 2012) were digested with XbaI (30 U for 3 h) (Takara, Bio, Otsu, Shiga, Japan) and subjected to PFGE as described elsewhere (Roderova et al 2016). The resulting restriction profiles of isolates were compared using the GelCompareII software (Applied Maths, Kortrijk, Belgium).…”