Molecular diversity of Extended-spectrum β-lactamase-producing Escherichia coli from vultures in Canary Islands

Abstract

Antimicrobial resistance among isolates from wild animals is increasingly reported. Extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae, and particularly Escherichia coli, have spread worldwide as one of the most common multidrug-resistant organisms. The aim of this study was to determine the carriage rate of ESBL-producing E. coli isolates and their genetic characteristics in wild vultures from the Canary Islands. Faecal samples were collected from 22 apparently healthy free-ranging (wild) vulture chicks from Lanzarote and Fuerteventura (Canary Islands) during July 2019. They were seeded in MacConkey agar supplemented with cefotaxime (2 μg ml^-1 ). Colonies with typical morphology of E. coli were identified by MALDI-TOF-MS. Antimicrobial susceptibility was done by disk diffusion. Phenotypic detection of ESBL was performed by double-disk tests. The presence of bla_CTX-M , bla_SHV , bla_TEM , bla_KPC and bla_OXA-48 genes, as well as mcr-1 (colistin resistance), tetA/tetB and int1 gene, was tested by PCR/sequencing. Phylogenetic groups and multilocus sequence typing (MLST) were determined by PCR/sequencing. ESBL-producing E. coli isolates were detected in 5/22 tested animals (22.7%), and all isolates (one/animal) carried bla_CTX-M genes: bla_CTX-M-15 (n = 3) and bla_CTX-M-55 (n = 2). ESBL-positive isolates were ascribed to phylogenetic group D (two isolates), B₁ (two isolates) and A (one isolate), and five sequence types were detected (ST/phylogenetic-group/ESBL): ST515/B1/CTX-M-15, ST1290/A/CTX-M-15, ST38/D/CTX-M-15, ST457/D/CTX-M-55 and ST6448/B₁ /CTX-M-55; this suggests a genetic diversity among these isolates. Three CTX-M-15-producing isolates contained the bla_TEM gene and one the tetA gene. To our knowledge, this appears to be the first report of ESBL-producing E. coli in vulture chicks from the Canary Islands.