Antibiotic resistance and virulence genes in nisin-resistant Enterococcus faecalis isolated from raw buffalo milk modulate the innate functions of rat macrophages

J Appl Microbiol. 2019 Sep;127(3):897-910. doi: 10.1111/jam.14343. Epub 2019 Jun 26.

Abstract

Aims: To elucidate the antibiotic resistance and virulence genes of nisin-resistant Enterococcus faecalis isolated from raw buffalo milk and to study the effect of nisin-sensitive and -resistant E. faecalis on the innate immunity of rats.

Methods and results: Slanetz-Bartley agar plates containing nisin were used to isolate nisin-resistant E. faecalis. The virulence factors were ascertained using quantitative real-time polymerase chain reaction. Cell viability, phagocytosis, intracellular survival and enzyme assays were performed to investigate the interaction of E. faecalis with rat macrophages. Nisin-resistant E. faecalis was less prone to phagocytosis and survived longer inside the macrophages, due to reduced production of reactive oxygen species and nitric oxide. The viability and activation of macrophages was also reduced in the presence of resistant E. faecalis, as observed by enhanced lactate dehydrogenase production and reduced β-galactosidase.

Conclusions: Nisin-resistant E. faecalis and its virulence factors were reported in raw buffalo milk. This study shows that nisin-resistant variants exhibited cross resistance to antibiotics and suppressed the innate immune responses of rats by directly affecting macrophage activity.

Significance and impact of the study: This study elucidated the contamination of raw buffalo milk by nisin-resistant E. faecalis, which may pose food safety risk.

Keywords: Enterococcus faecalis; antibiotics resistance; macrophage; nisin-resistance; virulence.

MeSH terms

  • Animals
  • Buffaloes
  • Drug Resistance, Bacterial / genetics
  • Enterococcus faecalis / drug effects
  • Enterococcus faecalis / genetics*
  • Enterococcus faecalis / isolation & purification
  • Macrophages / drug effects*
  • Macrophages / metabolism
  • Macrophages / physiology
  • Microbial Sensitivity Tests
  • Milk / microbiology*
  • Nisin / pharmacology
  • Rats
  • Reactive Oxygen Species / metabolism
  • Virulence / genetics
  • Virulence Factors / genetics

Substances

  • Reactive Oxygen Species
  • Virulence Factors
  • Nisin