The objective of this study was to evaluate the effect of clinical mastitis and (or) other diseases on reproductive performance in lactating Holstein cows. Cows (n=967) from a commercial dairy farm were divided into four groups retrospectively: cows with clinical mastitis and other diseases (MD, n=54), clinical mastitis only (M, n=154), other diseases only (D, n=187), and cows with no record of clinical mastitis or other diseases (H, n=572). Days in milk at first service (DIMFS), services per conception (S/C), days not pregnant (DNP), the rate at which animals became pregnant over time and the proportion of cows that remained non-pregnant during 224 days of lactation were evaluated. Groups MD and M had greater (P<0.05) DNP compared with H (155+/-15 and 140+/-5 vs. 88+/-2, respectively). Moreover, MD and M had greater (P<0.05) S/C compared with H (3.0+/-0.4 and 2.1+/-0.1 vs. 1.6+/-0.1, respectively). The rate at which animals became pregnant over time was less (P<0.05) for MD and M and tended (P=0.1) to be less for D when compared with H. In addition, proportion of cows that remained non-pregnant by 224 days of lactation was greater (P<0.05) in MD, M, and D compared with H. Cows with mastitis were also divided into three groups according to the day of occurrence of the first case of clinical mastitis: (1) clinical mastitis occurred before 56 days postpartum (MP1); (2) clinical mastitis occurred between 56 and 105 days after parturition (MP2); and (3) clinical mastitis occurred after 105 days postpartum (MP3) Regardless of the time of occurrence, DNP was greater (P<0.05) for cows with mastitis compared with H. Time of mastitis occurrence affected S/C in that cows in MP2 and MP3 had a greater S/C compared with H cows (P<0.05). Reproductive efficiency was decreased by the presence of clinical mastitis alone because a greater proportion of cows with mastitis remained non-pregnant over time. Moreover, a greater proportion of cows with mastitis or diseases remained non-pregnant by 224 postpartum. Furthermore, the negative effects on reproduction were exacerbated when cows experienced both clinical mastitis and other diseases.