Alterations in the proportions of peripheral blood lymphocyte subsets was monitored using flow cytometry, before, during and after the peripartum period (week-8 to week (wk) 16) of dairy cows, when pregnancy and lactational stresses prevail and disease incidence is highest. A health code was assigned to each animal at each sampling time, and a subset of non-pregnant, non-lactating (NPNL) cows were sampled for comparisons, and to examine the effect of pregnancy and lactation only, on lymphocyte subset proportions. Results were expressed as the percentage of positive stained cells expressing CD2, CD4, CD8, WC1(gamma delta), IgM and class II major histocompatibility complex (MHC-II) molecules, as determined by flow cytometry. Comparisons of subset proportions were made across time and between health status categories. Mean fluorescence intensity (MFI) values were also examined across the periparturient period for CD2, CD4, CD8 and gamma delta T cell subsets. All T cell subsets varied significantly during the perparturient period in healthy cows, especially around parturition. B cell and MHC-II+ populations remained relatively constant until after calving and then significantly (P < or = 0.06) decreased. Compared to NPNL cows, all T cell subsets were significantly lower in healthy cows at most time points, whereas B cell and MHC-II+ cells were comparable to the NPNL values. CD2, CD4, and CD8 subsets decreased significantly at wk-3 and returned to initial subset proportions by wk 16. Conversely, the WC1 population increased at wk-3 relative to calving but declined to approximate initial proportions at calving. Health status had no significant overall effect on any subset unless it was separated by weeks in which case there were interactions between health and week for CD2 and CD8. Results indicate that variations in T lymphocyte subsets and in the concentrations of surface marker molecules (MFI) occur more as a result of pregnancy and lactation than health status in the dairy cow and that these factors have the least influence upon B cell and MHC-II+ populations during the periparturient period.