Mammary Gland Pathology Subsequent to Acute Infection with Strong versus Weak Biofilm Forming Staphylococcus aureus Bovine Mastitis Isolates: A Pilot Study Using Non-Invasive Mouse Mastitis Model

PLoS One. 2017 Jan 27;12(1):e0170668. doi: 10.1371/journal.pone.0170668. eCollection 2017.

Abstract

Background: Biofilm formation by Staphylococcus aureus is an important virulence attribute because of its potential to induce persistent antibiotic resistance, retard phagocytosis and either attenuate or promote inflammation, depending upon the disease syndrome, in vivo. This study was undertaken to evaluate the potential significance of strength of biofilm formation by clinical bovine mastitis-associated S. aureus in mammary tissue damage by using a mouse mastitis model.

Methods: Two S. aureus strains of the same capsular phenotype with different biofilm forming strengths were used to non-invasively infect mammary glands of lactating mice. Biofilm forming potential of these strains were determined by tissue culture plate method, ica typing and virulence gene profile per detection by PCR. Delivery of the infectious dose of S. aureus was directly through the teat lactiferous duct without invasive scraping of the teat surface. Both bacteriological and histological methods were used for analysis of mammary gland pathology of mice post-infection.

Results: Histopathological analysis of the infected mammary glands revealed that mice inoculated with the strong biofilm forming S. aureus strain produced marked acute mastitic lesions, showing profuse infiltration predominantly with neutrophils, with evidence of necrosis in the affected mammary glands. In contrast, the damage was significantly less severe in mammary glands of mice infected with the weak biofilm-forming S. aureus strain. Although both IL-1β and TNF-α inflammatory biomarkers were produced in infected mice, level of TNF-α produced was significantly higher (p<0.05) in mice inoculated with strong biofilm forming S. aureus than the weak biofilm forming strain.

Conclusion: This finding suggests an important role of TNF-α in mammary gland pathology post-infection with strong biofilm-forming S. aureus in the acute mouse mastitis model, and offers an opportunity for the development of novel strategies for reduction of mammary tissue damage, with or without use of antimicrobials and/or anti-inflammatory compounds for the treatment of bovine mastitis.

MeSH terms

  • Animals
  • Biofilms / growth & development*
  • Cattle
  • Disease Models, Animal
  • Female
  • Interleukin-1beta / metabolism
  • Mammary Glands, Animal / metabolism
  • Mammary Glands, Animal / microbiology
  • Mammary Glands, Animal / pathology
  • Mastitis, Bovine / metabolism
  • Mastitis, Bovine / microbiology*
  • Mastitis, Bovine / pathology
  • Mice
  • Pilot Projects
  • Staphylococcal Infections / microbiology*
  • Staphylococcal Infections / pathology
  • Staphylococcus aureus / growth & development*
  • Staphylococcus aureus / metabolism
  • Staphylococcus aureus / pathogenicity
  • Tumor Necrosis Factor-alpha / metabolism

Substances

  • Interleukin-1beta
  • Tumor Necrosis Factor-alpha

Grants and funding

This work was supported by grants through the Australia India Strategic Research Fund [BF040038] from the Department of Innovation, Industry, Science and Research, Commonwealth Government of Australia (to TM), and the India-Australia Biotechnology Fund [BT/Indo-Aus./04/06/2009] from the Department of Biotechnology, Ministry of Science and Technology, Government of India (to NH). Thanks are also due to Curtin University for providing the International Postgraduate Research Scholarship and the Australian Postgraduate Award to JG-T in support of her doctoral studies. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."