Determination of antimicrobial resistance of Enterococcus strains isolated from pigs and their genotypic characterization by method of amplification of DNA fragments surrounding rare restriction sites (ADSRRS fingerprinting)

J Med Microbiol. 2017 Mar;66(2):175-183. doi: 10.1099/jmm.0.000400.

Abstract

Purpose: In this study, we analysed phenotypic resistance profiles and their reflection in the genomic profiles of Enterococcus spp. strains isolated from pigs raised on different farms.

Methodology: Samples were collected from five pig farms (n=90 animals) and tested for Enterococcus. MICs of 12 antimicrobials were determined using the broth microdilution method, and epidemiological molecular analysis of strains belonging to selected species (faecalis, faecium and hirae) was performed using the ADSRRS-fingerprinting (amplification of DNA fragments surrounding rare restriction sites) method with a few modifications.

Results: The highest percentage of strains was resistant to tetracycline (73.4 %), erythromycin and tylosin (42.5 %) and rifampin (25.2 %), and a large number of strains exhibited high-level resistance to both kanamycin (25.2 %) and streptomycin (27.6 %). The strains of E. faecalis, E. faecium and E. hirae (n=184) revealed varied phenotypic resistance profiles, among which as many as seven met the criteria for multidrug resistance (30.4 % of strains tested). ADSRRS-fingerprinting analysis produced 17 genotypic profiles of individual strains which were correlated with their phenotypic resistance profiles. Only E. hirae strains susceptible to all of the chemotherapeutics tested had two different ADSRRS profiles. Moreover, eight animals were carriers of more than one genotype belonging to the same Enterococcus spp., mainly E. faecalis.

Conclusion: Given the possibility of transmission to humans of the high-resistance/multidrug resistance enterococci and the significant role of pigs as food animals in this process, it is necessary to introduce a multilevel control strategy by carrying out research on the resistance and molecular characteristics of indicator bacterial strains isolated from animals on individual farms.

MeSH terms

  • Animals
  • Anti-Bacterial Agents / pharmacology
  • DNA Fingerprinting
  • DNA, Bacterial / genetics
  • DNA, Bacterial / isolation & purification*
  • Drug Resistance, Multiple, Bacterial*
  • Enterococcus faecalis / drug effects
  • Enterococcus faecalis / isolation & purification
  • Enterococcus faecium / drug effects
  • Enterococcus faecium / isolation & purification
  • Enterococcus hirae / drug effects
  • Enterococcus hirae / isolation & purification
  • Erythromycin / pharmacology
  • Genotyping Techniques*
  • Kanamycin / pharmacology
  • Microbial Sensitivity Tests
  • Nucleic Acid Amplification Techniques
  • Rifampin / pharmacology
  • Streptomycin* / pharmacology
  • Swine / microbiology*
  • Tetracycline / pharmacology
  • Tetracycline Resistance
  • Tylosin / pharmacology

Substances

  • Anti-Bacterial Agents
  • DNA, Bacterial
  • Kanamycin
  • Erythromycin
  • Tetracycline
  • Rifampin
  • Streptomycin
  • Tylosin