Genetic Characterization of vanA-Enterococcus faecium Isolates from Wild Red-Legged Partridges in Portugal

Vancomycin-resistant enterococci (VRE) have been detected in wild animals representing a public health concern. The red-legged partridge (Alectoris rufa) is a common game bird and its meat is consumed in several countries, including Portugal. Three hundred five fecal samples of red-legged partridge from the north of Portugal were screened for VRE. Samples were cultured on Slanetz-Bartley agar supplemented with vancomycin (4 mg/L) and six vanA-Enterococcus faecium were recovered. Isolates were tested for antibiotic resistance and virulence genes. Multilocus sequence typing (MLST) was performed to study the genotypic diversity of vanA-containing E. faecium. The six isolates showed erythromycin resistance and harbored the erm(B) gene and the four that were tetracycline resistant showed the tet(M) gene. The C-terminal region of the pbp5 gene of the ampicillin-resistant isolates (minimal inhibitory concentration range of 256 μg/ml) was sequenced. Two different pbp5 alleles were detected when considering the changes of amino acid in 461-629 region. All isolates harbored the esp gene, whereas hyl, together with the esp gene, was detected in five isolates. MLST analysis grouped the isolates as ST448 (n = 1), ST139 (n = 1), and ST18 (n = 4). Our findings show that the red-legged partridges could be a reservoir of antimicrobial resistance genes and may contribute to the dissemination and transference of the resistance genes to other animals and humans.