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

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 (CTXR) 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. CTXRE. 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 CTXRE. coli isolates (one/sample) were ESBL-producers (ESBL total rate: 7.8%) carrying the following ESBL genes: blaCTX-M-1 (n = 3), blaCTX-M-15 (n = 3), blaCTX-M-55 (n = 2), blaCTX-M-27 (n = 2) and blaCTX-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 CTXR 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.

Antimicrobial Resistance Genes and Diversity of Clones among Faecal ESBL-Producing Escherichia coli Isolated from Healthy and Sick Dogs Living in Portugal

“…Regarding the Portuguese situation, the prevalence of ESBL-producing E. coli isolates in healthy dogs obtained in this work is similar to previous studies performed in dogs and cats [ 12 , 22 , 23 ] in the South and the North of Portugal. Worldwide, this prevalence was lower than the ones obtained with faecal samples of healthy dogs in Germany, Brazil, or China (24–29%) [ 15 , 37 , 38 ], but it is similar to the results of previous studies performed in Tunisia and France (12.7–17%) [ 11 , 39 ].…”

“…In this study, the CMY-2 gene was the qAmpC β-lactamase type found among two CTX R -ESBL-negative isolates and one ESBL-producing isolate, and it has been previously reported among E. coli strains from healthy and sick pets worldwide [ 20 , 23 , 39 , 44 ]. The detection of tet A and/or tet B genes in most of our tetracycline-resistant isolates seem to be similar to the results obtained by Costa et al [ 45 ] from dogs, in Northern Portugal.…”

“…A total of 361 faecal samples were recovered from 127 healthy and 234 hospitalized dogs from different cities in Portugal. All samples were collected between April andAugust 2017 (one sample/animal) using standardized procedures [ 23 ].…”

Characterization of ESBL-Producing Escherichia coli and Klebsiella pneumoniae Isolated from Clinical Samples in a Northern Portuguese Hospital: Predominance of CTX-M-15 and High Genetic Diversity

“…In Germany, this enzyme was also the most prevalent ESBL type (around 50% of the strains investigated) [ 17 ]. Recently, the bla CTX-M-15 gene was the most frequently detected among E. coli of both sick and healthy dogs and cats in Portugal [ 14 , 15 ]. Nevertheless, the bla CTX-M-1 was the most common ESBL gene among healthy students in Portugal (Lisbon), followed by the bla CTX-M-15 gene [ 13 ].…”

“…Moreover, recent studies reported ESBL-producing K. pneumoniae isolates among urinary tract infections and invasive infections in patients in Portugal [ 4 , 5 , 9 ], as well as among healthy Portuguese students in Lisbon [ 13 ]. Furthermore, a previous report created by our group showed the presence of ESBL-producing E. coli among healthy and sick cats [ 14 ] and dogs [ 15 ] in Portugal.…”