Prevalence, Antimicrobial Resistance, and Genotypic Characterization of Vancomycin-Resistant Enterococci in Meat Preparations

Abstract

ABSTRACT A total of 160 samples of poultry (80), pork (40), and beef (40) preparations (red sausages, white sausages, hamburgers, meatballs, nuggets, minced meat, escalope, and crepes) were tested in northwestern Spain to determine the prevalence of vancomycin-resistant enterococci (VRE). VRE were detected in 38 (23.8%) samples (37.5% of poultry, 15.0% of pork, and 5.0% of beef samples). One strain per food sample was further characterized. Isolates were identified as Enterococcus faecium (14 strains), E. durans (10), E. hirae (7), E. gallinarum (5), and E. casseliflavus–E. flavescens (2). All strains showed resistance or intermediate susceptibility to three or more antimicrobials of clinical significance, in addition to vancomycin. High rates of resistance or intermediate susceptibility were observed for teicoplanin (81.6% of isolates), chloramphenicol (81.6%), erythromycin (100%), quinupristin-dalfopristin (89.5%), and ciprofloxacin (81.6%). A moderate rate of resistance or intermediate susceptibility emerged for ampicillin (34.2%) and tetracycline (36.8%). Genes encoding antimicrobial resistance and virulence were studied by PCR. The vanA, vanB, vanC-1, and vanC-2/3 genes were identified in 27, 1, 5, and 2 isolates, respectively. Other resistance genes or transposon sequences found were tet(L), tet(M), Tn5397 (tetracycline), erm(A), erm(B) (erythromycin), vat(D), and vat(E) (quinupristin-dalfopristin). Most isolates were free of virulence determinants (agg, hyl, and efaAfm genes were detected in one, one, and five strains, respectively). Strains were classified as not biofilm producers (crystal violet assay; 4 isolates) or weak biofilm producers (34 isolates). Cluster analysis (EcoRI ribotyping) suggested a strong genetic relationship among isolates from different types of meat preparations, animal species, and retail outlets. Meat preparations might play a role in the spread through the food chain of VRE with several resistance and virulence genes.