Diversity of vanA-type vancomycin-resistant Enterococcus faecium isolated from broilers, poultry slaughterers and hospitalized humans in Greece

This study investigated the prevalence of vancomycin-resistant enterococci (VRE) in the broiler production environment after the avoparcin ban and their epidemiological relationship with human clinical VRE from the same geographical regions in Greece. Caecal contents from broilers (n500) from eight livestock farms and faecal samples from poultry slaughterers (n50), all collected in two slaughterhouses during 200508, were analysed for species and vancomycin resistance gene identification using multiplex PCR. Sixty-three human clinical vancomycin-resistant Enterococcus faecium (VREF) isolates, obtained during 200609, were also examined. Discriminant analysis (DA) was used to establish the relationship of antimicrobial resistance profiles (ARPs) among broiler, poultry slaughterer and human clinical VREF. PFGE was conducted to study the genetic relatedness among VREF from the different sources. A total of 120 VRE were recovered from 113 (22.6) broiler samples. VREF carrying the vanA gene were predominant, being recovered from 72 (14.4) samples from five (62.5) broiler farms. Concerning poultry slaughterers, VREF were recovered from 10 (20) samples. Susceptibility testing revealed that broiler VREF were consistently resistant to tetracycline, whereas 93.7 of clinical VREF were resistant to ampicillin. Furthermore, 92.1 of clinical VREF compared with 54.4 of broiler VREF were multiresistant (resistant to at least five antimicrobial classes). DA classified broiler and human clinical VREF into their corresponding source with high classification rates (100 and 85.7, respectively), while the classification rate of poultry slaughterer VREF was relatively low (50), with 40 of them classified closely to broiler VREF. PFGE patterns were clearly related to the source of the VREF, with broiler isolates being clustered distinctly from all human isolates. A remarkable persistence of VREF was observed in the broiler production environment even 10 years after the avoparcin ban. Human and broiler VREF belonged to clearly unrelated populations, strongly indicating no clonal spread of VREF among the different sources, even between broilers and poultry slaughterers, despite them sharing common ARPs, as also supported by DA.