Antimicrobial Resistance Genes and Diversity of Clones among ESBL- and Acquired AmpC-Producing Escherichia coli Isolated from Fecal Samples of Healthy and Sick Cats in Portugal

Isabel Carvalho; Nadia Safia Chenouf; Rita Cunha; Carla Martins; Paulo Pimenta; Ana Raquel Pereira; Sandra Martínez-Álvarez; Sónia Ramos; Vanessa Silva; Gilberto Igrejas; Carmen Torres; Patrícia Poeta

doi:10.3390/antibiotics10030262

Antimicrobial Resistance Genes and Diversity of Clones among ESBL- and Acquired AmpC-Producing Escherichia coli Isolated from Fecal Samples of Healthy and Sick Cats in Portugal

Antibiotics (Basel). 2021 Mar 5;10(3):262. doi: 10.3390/antibiotics10030262.

Authors

Isabel Carvalho^{1

2

3

4

5}, Nadia Safia Chenouf^{5

6}, Rita Cunha⁷, Carla Martins⁸, Paulo Pimenta⁹, Ana Raquel Pereira¹⁰, Sandra Martínez-Álvarez⁵, Sónia Ramos¹¹, Vanessa Silva^{1

2

3

4}, Gilberto Igrejas^{2

3

4}, Carmen Torres⁵, Patrícia Poeta^{1

4}

Affiliations

¹ Microbiology and Antibiotic Resistance Team (MicroART), Department of Veterinary Sciences, University of Trás-os-Montes and Alto Douro, 5000-801 Vila Real, Portugal.
² Department of Genetics and Biotechnology, UTAD, 5000-801 Vila Real, Portugal.
³ Functional Genomics and Proteomics Unit, UTAD, 5000-801 Vila Real, Portugal.
⁴ Laboratory Associated for Green Chemistry (LAQV-REQUIMTE), New University of Lisbon, 2829-516 Monte da Caparica, Portugal.
⁵ Area Biochemistry and Molecular Biology, University of La Rioja, 26006 Logroño, Spain.
⁶ Laboratory of Exploration and Valuation of the Steppe Ecosystem, University of Djelfa, Djelfa 17000, Algeria.
⁷ Hospital Veterinário de São Bento, 1200-822 Lisboa, Portugal.
⁸ Clínica Veterinária do Vouga, 3740-253 Sever do Vouga, Portugal.
⁹ Hospital Veterinário de Trás-os-Montes, 5000-056 Vila Real, Portugal.
¹⁰ Centro Veterinário de Macedo de Cavaleiros, 5340 Bragança, Portugal.
¹¹ VetRedondo, Consultório Veterinário de Monte Redondo Unipessoal Lda, Monte Redondo, 2425-618 Leiria, Portugal.

Abstract

The aim of the study was to analyze the mechanisms of resistance in extended-spectrum beta-lactamase (ESBL)- and acquired AmpC (qAmpC)-producing Escherichia coli isolates from healthy and sick cats in Portugal. A total of 141 rectal swabs recovered from 98 sick and 43 healthy cats were processed for cefotaxime-resistant (CTX^R) E. coli recovery (in MacConkey agar supplemented with 2 µg/mL cefotaxime). The matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) method was used for E. coli identification and antimicrobial susceptibility was performed by a disk diffusion test. The presence of resistance/virulence genes was tested by PCR sequencing. The phylogenetic typing and multilocus sequence typing (MLST) were determined by specific PCR sequencing. CTX^RE. coli isolates were detected in seven sick and six healthy cats (7.1% and 13.9%, respectively). Based on the synergy tests, 11 of 13 CTX^RE. coli isolates (one/sample) were ESBL-producers (ESBL total rate: 7.8%) carrying the following ESBL genes: bla_CTX-M-1 (n = 3), bla_CTX-M-15 (n = 3), bla_CTX-M-55 (n = 2), bla_CTX-M-27 (n = 2) and bla_CTX-M-9 (n = 1). Six different sequence types were identified among ESBL-producers (sequence type/associated ESBLs): ST847/CTX-M-9, CTX-M-27, CTX-M-1; ST10/CTX-M-15, CTX-M-27; ST6448/CTX-M-15, CTX-M-55; ST429/CTX-M-15; ST101/CTX-M-1 and ST40/CTX-M-1. Three of the CTX^R isolates were CMY-2-producers (qAmpC rate: 2.1%); two of them were ESBL-positive and one ESBL-negative. These isolates were typed as ST429 and ST6448 and were obtained in healthy or sick cats. The phylogenetic groups A/B1/D/clade 1 were detected among ESBL- and qAmpC-producing isolates. Cats are carriers of qAmpC (CMY-2)- and ESBL-producing E. coli isolates (mostly of variants of CTX-M group 1) of diverse clonal lineages, which might represent a public health problem due to the proximity of cats with humans regarding a One Health perspective.

Keywords: CMY-2; CTX-M; E. coli; antimicrobial resistance; cats; extended spectrum beta-lactamase (ESBL), AmpC beta-lactamase; public health.

Grants and funding

SFRH/BD/137947/2018/Fundação para a Ciência e a Tecnologia