Methicillin-Resistant Staphylococcus aureus from Brazilian Dairy Farms and Identification of Novel Sequence Types

Zoonoses Public Health. 2016 Mar;63(2):97-105. doi: 10.1111/zph.12209. Epub 2015 Jul 14.

Abstract

The aim of this study was to investigate the phenotypic and genotypic diversity and anti-microbial resistance among staphylococci of dairy herds that originated from Paraiba State, north-eastern Brazil, a region where such studies are rare. Milk samples (n = 552) were collected from 15 dairy farms. Isolates were evaluated for anti-microbial susceptibility by Kirby-Bauer disc diffusion method. Confirmation of methicillin-resistant Staphylococcus aureus (MRSA) was performed using multiplex PCR targeting mecA and nuc genes in addition to phenotypic assay based on PBP-2a latex agglutination. Clonal relatedness of isolates was determined by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) genotyping. Staphylococci were detected in 269 (49%) of the samples. Among these, 65 (24%) were S. aureus. The remaining 204 isolates were either coagulase-negative staphylococci (n = 188; 70%) or coagulase positive other than S. aureus (n = 16; 6%). Staphylococci were cultured in seven (35%) of the 20 hand swab samples, from which five isolates were S. aureus. The isolates were most commonly resistant against penicillin (43%), ampicillin (38%) and oxacillin (27%). The gene mecA was detected in 21 S. aureus from milk and in one isolate from a milker's hand. None of the isolates were resistant to vancomycin. PFGE findings showed high clonal diversity among the isolates. Based on MLST, we identified a total of 11 different sequence types (STs 1, 5, 6, 83, 97, 126, 1583, 1622, 1623, 1624 and 1625) with four novel STs (ST1622-ST1625). The findings show that MRSA is prevalent in milk from semi-extensive dairy cows in north-eastern Brazil, and further investigation on its extent in various types of milk production systems and the farm-to-table continuum is warranted.

Keywords: Methicillin-resistant Staphylococcus aureus; milk; oxacillin resistance; oxacillin resistant Staphylococcus aureus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anti-Bacterial Agents / pharmacology
  • Bacterial Proteins / genetics
  • Biodiversity
  • Brazil / epidemiology
  • Cattle
  • Dairying
  • Drug Resistance, Multiple, Bacterial
  • Female
  • Food Contamination
  • Food Microbiology
  • Genetic Variation
  • Genotype
  • Humans
  • Methicillin-Resistant Staphylococcus aureus / classification
  • Methicillin-Resistant Staphylococcus aureus / genetics*
  • Methicillin-Resistant Staphylococcus aureus / isolation & purification*
  • Microbial Sensitivity Tests
  • Micrococcal Nuclease / genetics
  • Milk / microbiology*
  • Multilocus Sequence Typing
  • Penicillin-Binding Proteins / genetics
  • Phenotype
  • Polymerase Chain Reaction
  • Staphylococcal Food Poisoning / epidemiology
  • Staphylococcal Food Poisoning / veterinary*
  • Staphylococcus aureus / drug effects

Substances

  • Anti-Bacterial Agents
  • Bacterial Proteins
  • Penicillin-Binding Proteins
  • mecA protein, Staphylococcus aureus
  • nuc protein, staphylococcus
  • Micrococcal Nuclease