Detection and genetic characterization of blaESBL-carrying plasmids of cloacal Escherichia coli isolates from white stork nestlings (Ciconia ciconia) in Spain

Sandra Martínez-Álvarez; Pierre Châtre; Teresa Cardona-Cabrera; Pauline François; Alberto Sánchez-Cano; Ursula Höfle; Myriam Zarazaga; Jean-Yves Madec; Marisa Haenni; Carmen Torres

doi:10.1016/j.jgar.2023.07.011

Detection and genetic characterization of bla_ESBL-carrying plasmids of cloacal Escherichia coli isolates from white stork nestlings (Ciconia ciconia) in Spain

J Glob Antimicrob Resist. 2023 Sep:34:186-194. doi: 10.1016/j.jgar.2023.07.011. Epub 2023 Jul 22.

Affiliations

¹ Area of Biochemistry and Molecular Biology, OneHealth-UR Research Group, University of La Rioja, Logroño, Spain.
² ANSES - Université de Lyon, Unité Antibiorésistance et Virulence Bactériennes, Lyon, France.
³ Health and Biotechnology (SaBio) Research Group, Institute for Game and Wildlife Research IREC (CSIC-UCLM), Ciudad Real, Spain.
⁴ Area of Biochemistry and Molecular Biology, OneHealth-UR Research Group, University of La Rioja, Logroño, Spain. Electronic address: carmen.torres@unirioja.es.

PMID: 37482121
DOI: 10.1016/j.jgar.2023.07.011

Abstract

Objectives: This study aimed to characterize Escherichia coli isolates from cloacal samples of white stork nestlings, with a special focus on extended-spectrum β-lactamases (ESBLs)-producing E. coli isolates and their plasmid content.

Methods: Cloacal samples of 88 animals were seeded on MacConkey-agar and chromogenic-ESBL plates to recover E. coli and ESBL-producing E. coli. Antimicrobial susceptibility was screened using the disc diffusion method, and the genotypic characterization was performed by polymerase chain reaction (PCR) and subsequent sequencing. S1 nuclease Pulsed-Field-Gel-Electrophoresis (PFGE), Southern blotting, and conjugation essays were performed on ESBL-producing E. coli, as well as whole-genome sequencing by short- and long-reads. The four bla_ESBL-carrying plasmids were completely sequenced.

Results: A total of 113 non-ESBL-producing E. coli isolates were collected on antibiotic-free MacConkey-agar, of which 27 (23.9%) showed a multidrug-resistance (MDR) phenotype, mainly associated with β-lactam-phenicol-sulfonamide resistance (bla_TEM/cmlA/floR/sul1/sul2/sul3). Moreover, four white stork nestlings carried ESBL-producing E. coli (4.5%) with the following characteristics: bla_SHV-12/ST38-D, bla_SHV-12/ST58-B1, bla_CTX-M-1/ST162-B1, and bla_CTX-M-32/ST155-B1. Whole-genome sequencing followed by Southern blot hybridizations on S1-PFGE gels in ESBL-positive isolates proved that the bla_CTX-M-1 gene and one of the bla_SHV-12 genes were carried by IncI1/pST3 plasmids, while the second bla_SHV-12 gene and the bla_CTX-M-32 gene were located on IncF plasmids. The two bla_SHV-12 genes and the two bla_CTX-M genes had similar but non-identical close genetic environments, as all four genes were flanked by a variety of insertion sequences.

Conclusion: The role played by several genetic platforms in the mobility of ESBL genes allows for interchangeability on a remarkably small scale (gene-plasmid-clones), which may support the spread of ESBL genes.

Keywords: ESBL; Integrons; Plasmids; Transfer technique; White stork nestlings.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Agar
Animals
Birds* / microbiology
Escherichia coli Infections*
Escherichia coli* / genetics
Escherichia coli* / isolation & purification
Plasmids / genetics
Spain
beta-Lactamases / genetics

Substances

Agar
beta-Lactamases