Paternal care is associated with a reduced likelihood of engaging in competitive or mating behavior and an increased likelihood of providing protection when necessary. Over recent years, there has been increasing evidence to assume that the steroid testosterone (T) in men might reflect the degree of mating effort. In line with this, decreased T levels were shown in fathers compared to non-fathers and it was suggested that paternal care, and most behavior positively associated with T, might be incompatible with each other. Independently, the personality trait sensation seeking (SS) has been related to mating behavior and also to elevated T in men. Aiming to integrate these different lines of research in a longitudinal approach, we explored the impact of SS on T levels in the context of the transition to fatherhood. Thirty-seven fathers and 38 men without children but in committed, romantic relationships (controls) were recruited. At two time points (for fathers: four weeks prior to (t1) and eight weeks after birth (t2)), all subjects repeatedly collected saliva samples for T measurement, filled in a protocol of activities during the course of these days and completed an online questionnaire. In line with our hypotheses, the results show significantly lower aggregated (AUC-T) T levels in fathers compared with non-fathers. Furthermore, moderation analyses revealed a significant interaction between group and SS at t2, with the lowest T levels in low SS fathers. These data suggest that adaptation processes of the transition to fatherhood are influenced by individual differences in personality traits.