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Genetic characterisation of extended-spectrum β-lactamases in Escherichia coli isolated from retail chicken products including CTX-M-9 containing isolates: a food safety risk factor
Authors:N Silva  L Costa  A Gonçalves  M Sousa  H Radhouani  F Brito
Institution:1. Centre of Studies of Animal and Veterinary Sciences , Vila Real , Portugal;2. Centre of Studies of Animal and Veterinary Sciences , Vila Real , Portugal;3. Veterinary Science Department , University of Trás-os-Montes and Alto Douro , Vila Real;4. Institute for Biotechnology and Bioengineering, Centre of Genomics and Biotechnology, University of Trás-os-Montes and Alto Douro;5. Department of Genetics and Biotechnology , University of Trás-os-Montes and Alto Douro , Vila Real , Portugal;6. Veterinary Science Department , University of Trás-os-Montes and Alto Douro , Vila Real
Abstract:1. Bacterial resistance to β-lactam antibiotics has risen dramatically in Escherichia coli from food animals. In a previous study, 29 randomly selected chicken products, collected in Portugal, were analysed for the presence of extended-spectrum β-lactamases (ESBLs)-producing E. coli; and during this study the genetic characterisation of ESBLs genes was investigated.

2. The presence of genes encoding TEM, OXA, SHV, and CTX-M type beta-lactamases was studied by PCR followed by sequencing. Additionally, other mechanisms of antimicrobial resistance, phylogenetic groups and the presence of virulence determinants were evaluated among the isolates.

3. β-lactamases genes were identified as follows: bla CTX-M-14 (n?=?4), bla CTX-M-1 (n?=?2), bla CTX-M-9 (n?=?4) and bla TEM-52 (n?=?13). Mutations at positions ?42, ?18, ?1, and +58 of ampC promoter region were identified in 4 non-ESBL-producing isolates. The tet(A) or tet(B) genes were identified in all tetracycline-resistant isolates; the aadA gene detected in 8 of 10 streptomycin-resistant isolates; the aac(3)-II gene in all gentamicin-resistant isolates; the cmlA gene in the chloramphenicol-resistant isolate; and sul1 and/or sul2 and/or sul3 genes were found in all trimethoprim-sulfamethoxazole-resistant isolates. The intI1 gene was detected in 8 trimethoprim-sulfamethoxazole-resistant isolates and the intI2 gene in 4 isolates; one gene cassette arrangements were identified among class 1 integrons (dfrA1?+?aadA1) and among the class 2 integrons (dfrA1?+?sat2?+?aadA1). Among cefotaxime-resistant isolates, 16 belonged to A or B1 phylogenetic groups, while 11 isolates were classified into the D or B2 phylogroups. At least one virulence-associated gene (aer, fimA, or papC) was detected in 74·1% of the cefotaxime-resistant isolates.

4. Because ESBLs-producing bacteria are resistant to a broad range of β-lactams, infections caused by these organisms complicate therapy and limit treatment options.
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