Enterotoxin production and antimicrobial susceptibility in Staphylococci isolated from traditional raw milk cheeses in Serbia

J Environ Sci Health B. 2017 Dec 2;52(12):864-870. doi: 10.1080/03601234.2017.1361764. Epub 2017 Sep 26.

Abstract

This study was undertaken to determine the prevalence of coagulase positive staphylococci (CPS) by examining a total of 71 raw milk cheeses. Additionally, enterotoxigenicity, antimicrobial susceptibility and the presence of mecA and mecC genes in the staphylococcal isolates were investigated. The isolation and enumeration procedure of CPS followed the International Organization for Standardization (ISO) standard. The presumptive staphylococci were identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) using the VITEK MS system. VIDAS® Staph enterotoxin II assay was used for the detection of classical enterotoxins. Antimicrobial susceptibility testing (AST) was accomplished performing the disk diffusion method. All suspected methicillin resistant staphylococci were investigated for the presence of mecA and mecC genes by PCR assay. A high prevalence (87.32%) of CPS was detected in the cheeses at contamination levels up to 5.58 log CFU g-1. Among 47 staphylococcal isolates screened for enterotoxin production, only one isolate, identified as S. hyicus, was confirmed as being enterotoxigenic. Resistance to penicillin (63.70%) was the most common resistance among the tested Staphylococcus aureus isolates. The dominant phenotypic resistance patterns in coagulase negative staphylococci (CNS) were resistance to ofloxacin and fusidic acid. All CNS isolates were susceptible to the clinically important antibiotics clindamycin, chloramphenicol, gentamicin, linezolid, rifampicin and trimethoprim-sulfamethoxazole. The mecA and mecC genes were not detected. To the best of our knowledge, this is the first study concerning evaluation of the presence of methicillin resistant staphylococci (MRS) in dairy products in Serbia.

Keywords: Antimicrobial resistance; Serbian raw milk cheeses; enterotoxin production; staphylococci.

MeSH terms

  • Animals
  • Anti-Bacterial Agents / pharmacology
  • Bacterial Proteins / genetics
  • Cheese / microbiology*
  • Coagulase / metabolism
  • Drug Resistance, Bacterial / drug effects
  • Enterotoxins / genetics
  • Enterotoxins / metabolism
  • Methicillin-Resistant Staphylococcus aureus / isolation & purification
  • Microbial Sensitivity Tests
  • Penicillin-Binding Proteins / genetics
  • Polymerase Chain Reaction
  • Serbia
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Staphylococcus / drug effects*
  • Staphylococcus / genetics
  • Staphylococcus / isolation & purification
  • Staphylococcus / metabolism*

Substances

  • Anti-Bacterial Agents
  • Bacterial Proteins
  • Coagulase
  • Enterotoxins
  • Penicillin-Binding Proteins
  • mecA protein, Staphylococcus aureus