The distribution of Staphylococcus aureus within herds seems to be related to interactions among the shedding characteristics of the bacteria, their pathogenicity and mammary gland immune status. The aim of the present study was to investigate the relationships between selected mammary gland immune factors and intramammary infections associated with Staph. aureus. Overall, 70 cows from five commercial dairy herds were included in the study and quarter milk samples were assessed using bacteriological and cytological tests. We evaluated differential cell count, lysozyme concentration, N-acetyl-β-glucosaminidase (NAGase) activity, cell viability and respiratory burst activity in randomly chosen quarter milk samples from each cow. Staph. aureus intramammary infection elicited different responses in the mammary gland immune defences investigated. Polymorphonuclear leucocytes (PMN) as a proportion of total somatic cells in milk, cell viability and NAGase activity were higher in infected quarters, while the proportions of macrophages and lymphocytes, respiratory burst activity and lysozyme levels were lower. Mean values differed among herds, but the differences were not significant. These changes were associated with Staph. aureus infection. The reduced respiratory burst activity together with the increase in the proportion of PMN suggests that both the number and activity of PMN could influence the susceptibility of the mammary gland to pathogens. Indeed, the logistic model adopted suggests that impairment of milk immune factors could be concurrent with the development of an infection.