Antimicrobial Resistance Genes and Diversity of Clones among Faecal ESBL-Producing <i>Escherichia coli</i> Isolated from Healthy and Sick Dogs Living in Portugal
Isabel Carvalho,
Rita Cunha,
Carla Martins,
Sandra Martínez-Álvarez,
Nadia Safia Chenouf,
Paulo Pimenta,
Ana Raquel Pereira,
Sónia Ramos,
Madjid Sadi,
Ângela Martins,
Jorge Façanha,
Fazle Rabbi,
Rosa Capita,
Carlos Alonso-Calleja,
Maria de Lurdes Nunes Enes Dapkevicius,
Gilberto Igrejas,
Carmen Torres,
Patrícia Poeta
Affiliations
Isabel Carvalho
Microbiology and Antibiotic Resistance Team (MicroART), Department of Veterinary Sciences, University of Trás-os-Montes and Alto Douro, 5000-801 Vila Real, Portugal
Rita Cunha
Hospital Veterinário Cascais da Onevet, 2775-352 Parede, Lisbon, Portugal
Carla Martins
Clínica Veterinária do Vouga, 3740-253 Sever do Vouga, Portugal
Sandra Martínez-Álvarez
Area Biochemistry and Molecular Biology, University of La Rioja, 26006 Logroño, Spain
Nadia Safia Chenouf
Area Biochemistry and Molecular Biology, University of La Rioja, 26006 Logroño, Spain
Paulo Pimenta
Hospital Veterinário de Trás-os-Montes, 5000-056 Vila Real, Portugal
Ana Raquel Pereira
Centro Veterinário de Macedo de Cavaleiros, 5340-202 Bragança, Portugal
Sónia Ramos
VetRedondo, Consultório Veterinário de Monte Redondo Unipessoal Lda, Monte Redondo, 2425-618 Leiria, Portugal
Madjid Sadi
Area Biochemistry and Molecular Biology, University of La Rioja, 26006 Logroño, Spain
Ângela Martins
Animal and Veterinary Research Center (CECAV), University of Trás-os-Montes and Alto Douro, 5000-801 Vila Real, Portugal
Jorge Façanha
Centro Veterinário Jorge Façanha, 5140-060 Carrazeda de Ansiães, Portugal
Fazle Rabbi
Australian Computer Society, Docklands, Melbourne, VIC 3008, Australia
Rosa Capita
Department of Food Hygiene and Technology, Veterinary Faculty, University of León, 24071 León, Spain
Carlos Alonso-Calleja
Department of Food Hygiene and Technology, Veterinary Faculty, University of León, 24071 León, Spain
Maria de Lurdes Nunes Enes Dapkevicius
Faculty of Agricultural and Environmental Sciences, University of the Azores, 9500-321 Angra do Heroísmo, Portugal
Gilberto Igrejas
Department of Genetics and Biotechnology, University of Trás-os-Montes and Alto Douro, 5000-801 Vila Real, Portugal
Carmen Torres
Area Biochemistry and Molecular Biology, University of La Rioja, 26006 Logroño, Spain
Patrícia Poeta
Microbiology and Antibiotic Resistance Team (MicroART), Department of Veterinary Sciences, University of Trás-os-Montes and Alto Douro, 5000-801 Vila Real, Portugal
The purpose of this study was to analyse the prevalence and genetic characteristics of ESBL and acquired-AmpC (qAmpC)-producing Escherichia coli isolates from healthy and sick dogs in Portugal. Three hundred and sixty-one faecal samples from sick and healthy dogs were seeded on MacConkey agar supplemented with cefotaxime (2 µg/mL) for cefotaxime-resistant (CTXR) E. coli recovery. Antimicrobial susceptibility testing for 15 antibiotics was performed and the ESBL-phenotype of the E. coli isolates was screened. Detection of antimicrobial resistance and virulence genes, and molecular typing of the isolates (phylogroups, multilocus-sequence-typing, and specific-ST131) were performed by PCR (and sequencing when required). CTXRE. coli isolates were obtained in 51/361 faecal samples analysed (14.1%), originating from 36/234 sick dogs and 15/127 healthy dogs. Forty-seven ESBL-producing E. coli isolates were recovered from 32 sick (13.7%) and 15 healthy animals (11.8%). Different variants of blaCTX-M genes were detected among 45/47 ESBL-producers: blaCTX-M-15 (n = 26), blaCTX-M-1 (n = 10), blaCTX-M-32 (n = 3), blaCTX-M-55 (n = 3), blaCTX-M-14 (n = 2), and blaCTX-M-variant (n = 1); one ESBL-positive isolate co-produced CTX-M-15 and CMY-2 enzymes. Moreover, two additional CTXR ESBL-negative E. coli isolates were CMY-2-producers (qAmpC). Ten different sequence types were identified (ST/phylogenetic-group/β-lactamase): ST131/B2/CTX-M-15, ST617/A/CTX-M-55, ST3078/B1/CTX-M-32, ST542/A/CTX-M-14, ST57/D/CTX-M-1, ST12/B2/CTX-M-15, ST6448/B1/CTX-M-15 + CMY-2, ST5766/A/CTX-M-32, ST115/D/CMY-2 and a new-ST/D/CMY-2. Five variants of CTX-M enzymes (CTX-M-15 and CTX-M-1 predominant) and eight different clonal complexes were detected from canine ESBL-producing E. coli isolates. Although at a lower rate, CMY-2 β-lactamase was also found. Dogs remain frequent carriers of ESBL and/or qAmpC-producing E. coli with a potential zoonotic role.
