Integrative phenotypic and genomic analysis of extended-spectrum Beta-lactamase (ESBL) and carbapenemase genes in Enterobacteriaceae and Pseudomonaceae strains isolated from animals in a Spanish Veterinary Teaching Hospital

Laura Leal-Vélez; Sergio Quevedo-Caraballo; Raffaele Scarpellini; Marta E García; José L Blanco

doi:10.1016/j.rvsc.2025.105529

Integrative phenotypic and genomic analysis of extended-spectrum Beta-lactamase (ESBL) and carbapenemase genes in Enterobacteriaceae and Pseudomonaceae strains isolated from animals in a Spanish Veterinary Teaching Hospital

Res Vet Sci. 2025 Jan 8:185:105529. doi: 10.1016/j.rvsc.2025.105529. Online ahead of print.

Authors

Laura Leal-Vélez¹, Sergio Quevedo-Caraballo², Raffaele Scarpellini³, Marta E García², José L Blanco²

Affiliations

¹ Department of Animal Health, Faculty of Veterinary Medicine, Complutense University of Madrid, Av. Puerta de Hierro s/n, 28040 Madrid, Spain; Complutense University of Madrid, Faculty of Veterinary Medicine, Veterinary Teaching Hospital, av. Puerta de Hierro s/n, 28040 Madrid, Spain. Electronic address: lauralea@ucm.es.
² Department of Animal Health, Faculty of Veterinary Medicine, Complutense University of Madrid, Av. Puerta de Hierro s/n, 28040 Madrid, Spain; Complutense University of Madrid, Faculty of Veterinary Medicine, Veterinary Teaching Hospital, av. Puerta de Hierro s/n, 28040 Madrid, Spain.
³ Department of Veterinary Medical Sciences, Alma Mater Studiorum, University of Bologna, Via Tolara di Sopra 50, Ozzano Emilia, BO 40006, Italy.

PMID: 39808975
DOI: 10.1016/j.rvsc.2025.105529

Abstract

Antimicrobial resistance (AMR) is a major global health threat, exacerbated by globalization which facilitates the spread of resistant bacteria. Addressing this issue requires a One Health perspective, involving humans, animals, and the environment. This study aims to compare the phenotypic resistance profiles of 69 clinical bacterial isolates (Enterobacteriaceae and Pseudomonaceae) from a Veterinary Teaching Hospital in Spain with their genotypic resistance profiles based on the presence of Extended-Spectrum Beta-Lactamases (ESBLs), AmpC and carbapenemases -enconding genes. For the genotypical analysis, whole genome sequencing (WGS) was used. Phenotypic characterization revealed that 37 isolates (53.6 %) grew on ESBL-selective medium. Phenotypic confirmatory tests showed that 12 strains (17.4 %) had some type of ESBL and 21 (30.4 %) could have an AmpC. Also, 24 isolates (34.8 %) grew in selective media for carbapenemases-producing bacteria, and 2 of these had a class A carbapenemase based on the KPC&MBL&OXA-48 disc kit. The genotypic analysis revealed 20 isolates (29 %) had bla_TEM, 8 (11.6 %) had bla_CTX-M and 7 (10.1 %) bla_SHV. 27 (39.1 %) isolates had class C beta-lactamase genes. 35 isolates (50.7 %) had bla_OXA, class D beta-lactamase. 37 strains (53.6 %) had an Inc. plasmid replicon associated with the spread of AMR genes, including beta-lactamases and carbapenemases. This study emphasizes the value of combining phenotypic and genomic analyses to better understand and address antibiotic resistance, especially in veterinary contexts. Integrating these approaches enhances diagnostic accuracy by identifying strains with resistance genes that may not show phenotypically, helping clinicians in anticipating resistance under selective pressure.

Keywords: Antimicrobial resistance; Carbapenemase; ESBL; Enterobacteriaceae; Pseudomonaceae; Whole genome sequencing.