Free Access
Vet. Res.
Volume 36, Number 4, July-August 2005
Page(s) 629 - 643
How to cite this article Vet. Res. (2005) 629-643
Vet. Res. 36 (2005) 629-643
DOI: 10.1051/vetres:2005023

Neutrophil apoptosis during experimentally induced Staphylococcus aureus mastitis

Zbysek Sladeka, Dusan Rysanekb, Helena Ryznarovaa and Martin Faldynab

a  Department of Morphology, Physiology and Veterinary Sciences, Mendel University of Agriculture and Forestry, Zemedelska 1, 613 00 Brno, Czech Republic
b  Veterinary Research Institute, Hudcova 70, 621 32 Brno, Czech Republic

(Received 8 October 2004; accepted 14 January 2005)

Abstract - The objective of this study was to determine whether neutrophil apoptosis and their consequent elimination by macrophages from the mammary gland is modulated by an infection caused by Staphylococcus aureus (S. aureus). The study was performed on twenty mammary glands of 5 virgin heifers. A buffered physiological solution (PBS) was administered as a means of control into the mammary glands of the heifers and after 168 h, the glands were inoculated with S. aureus. The samples of cell populations were obtained by lavages of the mammary glands in 4 intervals (24, 48, 72 and 168 h) after the experimental infection. Flow cytometry was used for determination of Annexin-V positivity and propidium iodide (PI) negativity of neutrophils. Light microscopy was used for determination of neutrophil karyopyknosis. Cytochemistry was used for the detection of myeloperoxidase-positive (MPO+) macrophages. Instillation of S. aureus resulted in an intramammary infection which persisted during the following experimental period. The total number of both Annexin-V-positive and PI negative neutrophils and karyopyknotic neutrophils peaked at 24 h after both of PBS and S. aureus administration. The highest percentages of Annexin-V-positive and PI negative neutrophils and karyopyknotic neutrophils were detected 48 and 168 h after PBS and S. aureus administration, respectively. The total number of MPO+ macrophages was the highest 24 h and 48 h after PBS and S. aureus administration, respectively; the percentage of MPO+ macrophages was the highest at 72 h in both cases. The dynamics of resolution of mastitis caused by S. aureus was very similar to the resolution of inflammatory response of the mammary gland after PBS administration. Mechanisms of cell pathogen elimination as well as inflammation resolution were very intensively involved; nevertheless, the mammary gland infection persisted. An early inclusion of the mechanisms of an acute inflammatory resolution thus paradoxically led to chronic infection.

Key words: Staphylococcus aureus / mastitis / neutrophil apoptosis

Corresponding author: Zbysek Sladek

© INRA, EDP Sciences 2005