Extended-Spectrum Beta-Lactamase-Producing Klebsiella pneumoniae Isolated from Healthy and Sick Dogs in Portugal

Isabel Carvalho; Carla Andrea Alonso; Vanessa Silva; Paulo Pimenta; Rita Cunha; Carla Martins; Gilberto Igrejas; Carmen Torres; Patrícia Poeta

doi:10.1089/mdr.2019.0205

Extended-Spectrum Beta-Lactamase-Producing Klebsiella pneumoniae Isolated from Healthy and Sick Dogs in Portugal

Microb Drug Resist. 2020 Jun;26(6):709-715. doi: 10.1089/mdr.2019.0205. Epub 2019 Dec 31.

Authors

Isabel Carvalho^{1

2

3

4}, Carla Andrea Alonso⁵, Vanessa Silva^{1

2

3

4}, Paulo Pimenta⁶, Rita Cunha⁷, Carla Martins⁸, Gilberto Igrejas^{2

3

4}, Carmen Torres⁵, Patrícia Poeta^{1

4}

Affiliations

¹ MicroART-Antibiotic Resistance Team, Department of Veterinary Sciences, Universidade de Trás-os-Montes e Alto Douro (UTAD), Vila Real, Portugal.
² Department of Genetics and Biotechnology, UTAD, Vila Real, Portugal.
³ Functional Genomics and Proteomics Unit, UTAD, Vila Real, Portugal.
⁴ Associated Laboratory for Green Chemistry (LAQV-REQUIMTE), University NOVA of Lisbon, Caparica, Portugal.
⁵ Area Biochemistry and Molecular Biology, University of La Rioja, Logroño, Spain.
⁶ Veterinary Hospital of Trás-os-Montes, Vila Real, Portugal.
⁷ Veterinary Hospital of São Bento, Lisbon, Portugal.
⁸ Veterinary Clinic of Vouga, Sever do Vouga, Portugal.

PMID: 31895642
DOI: 10.1089/mdr.2019.0205

Abstract

Extended-spectrum beta-lactamase (ESBL)- and carbapenemase (CP)-producing Klebsiella pneumoniae isolates are a public health concern at clinical level, mainly in Southern European countries. However, there are scarce data on the role of companion animals in the emergence of resistance to clinically relevant antibiotics. Therefore, our study aimed to determine the presence of K. pneumoniae with relevant beta-lactamases in fecal samples from healthy dogs (kennel and house dogs) and sick dogs in seven different hospitals in Portugal. Fecal samples from 125 healthy dogs and 231 sick dogs (one per animal) were collected during April-August 2017. Samples were screened on MacConkey agar supplemented with meropenem, and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) was used for K. pneumoniae identification. Genotypic detection of ESBLs or CPs was carried out by PCR/sequencing. Moreover, the presence of other antimicrobial resistance genes and multilocus sequence typing was tested by PCR/sequencing. K. pneumoniae isolates were obtained from 16 tested samples (4.4%), and 3 of them were ertapenem and/or meropenem intermediate/resistant (all of them imipenem susceptible and negative for CP genes). Fifteen K. pneumoniae isolates were ESBL producers, and they carried the following beta-lactamase genes: bla_CTX-M-15+bla_SHV-28 (four isolates, in three cases associated with bla_TEM-1), bla_CTX-M-15+bla_SHV-1 (five isolates, associated with TEM-1 in three cases), and bla_SHV-28+bla_TEM-1 (six isolates). Three ESBL-producing K. pneumoniae isolates of different origins and beta-lactamase genotypes (CTX-M-15+SHV-28, CTX-M-15+SHV-28+TEM-1, or SHV-28+TEM-1) belonged to the lineage ST307, and one isolate was identified as ST15 (CTX-M-15+SHV-1). These findings highlight that dogs are frequent carriers of ESBL-producing K. pneumonia isolates, harboring mostly genes encoding CTX-M-15 or SHV-28, associated in some cases with the high-risk clones ST307 and ST15.

Keywords: CTX-M-15; Klebsiella pneumoniae; SHV-28; ST15; ST307; dogs.

MeSH terms

Animals
Anti-Bacterial Agents / pharmacology
DNA, Bacterial
Dog Diseases / epidemiology
Dog Diseases / microbiology
Dogs
Drug Resistance, Multiple, Bacterial / genetics*
Klebsiella Infections / epidemiology
Klebsiella Infections / veterinary
Klebsiella pneumoniae / drug effects*
Klebsiella pneumoniae / genetics*
Klebsiella pneumoniae / isolation & purification
Polymerase Chain Reaction
Portugal
Sequence Analysis, DNA
beta-Lactam Resistance / genetics*

Substances

Anti-Bacterial Agents
DNA, Bacterial