Reproduction in most fish is typically a seasonal process, as spawning takes place usually at a given time of the year, depending on the reproduction strategy of the species, to ensure maximal survival of offspring. Nevertheless, fish reproduction cannot be considered an exclusively annual phenomenon, because spawning may also show daily rhythmicity. In this study, we investigated the existence of a daily spawning rhythm in gilthead seabream (Sparus aurata L) exposed to different light-dark (LD) cycles and at different times of the year using an automatic and programmable egg collector. Floatability and fertilization rates were analyzed at different times throughout the 24 h. The results showed a daily spawning rhythm with spanning occurring from 14:30 to 21:30 h, with the acrophase (peak time) being 18:29 and 18:08 h in fish exposed to an artificial photoperiod of 9L:15D in winter and in spring, respectively. Nevertheless, in fish exposed to a natural photoperiod of 12L:12D in spring, the acrophase of the rhythm was recorded later, at 21:28 h. The average fertilization rate was 87%, and no significant differences were found between the different hours of spawning. Moreover, when the LD cycle (9L:15D) was shifted by 12 h, the daily spawning rhythm gradually re-synchronized, resuming a stable phase-relationship after 4-5 transient days, which is characteristic of a endogenous circadian rhythm. Our results clearly demonstrated the existence of a 24 h period of spawning in gilthead sea bream, with a peak anticipating the forthcoming night, and its capacity to gradually re-synchronize after a 12 h shift in the LD cycle, pointing to the endogenous nature of this rhythm. These findings will be valuable for better understanding the reproductive physiology of this species and for optimizing the protocols of egg collection and larvae production in aquaculture.