ABSTRACT The aim of this study was to evaluate the effects of diphenyl diselenide (PhSe) supplementation during the transition period on the metabolism, immunity, oxidative status, and milk production of postpartum dairy cows. Twenty-seven Holstein females were divided into groups: (PhSe) (DDG), which received 3 μmol/kg of (PhSe)2 in 4 mL of dimethyl sulfoxide (DMSO) subcutaneously, DMSO (DMSOG), and NaCl (NACLG) which received 4 mL of DMSO and 0.9% NaCl. Evaluation of body condition score (BCS), weighing and administrations were performed at 42, 28, and 14 days prepartum, and on the day of calving (0). On days 0, 7, 14, 21, and 35 postpartum, BCS was evaluated, and blood were collected. Colostrum was obtained from the first postpartum milking. Production was measured, and milk was collected on days 7, 14, 21, and 35 postpartum. DDG showed higher values of total protein (TP) and globulins (GLOB) 7 days postpartum and increase from day 0 to days 21 and 35. DDG immunoglobulin G (IgG) was higher on days 21 and 35, and increased between days 7 and 21. In postpartum TP, GLOB and IgG are physiologically reduced, therefore, the results indicate that supplementation of transition females with (PhSe) stimulated postpartum humoral immunity.