Changes in different immune activities in the skin mucus of gilthead seabream (Sparus aurata L.) and European sea bass (Dicentrarchus labrax L.) specimens exposed to a constant light–dark photoperiod (12 h L:12 h D) were studied. Samples were collected at 08:00 (light on), 14:00, 20:00 (light off), 02:00, and again at 08:00 to determine immunoglobulin M (IgM) levels, several enzymes related to the immune system, and bactericidal activity. IgM levels were higher during the day in seabream and reached a minimum value at 20:00, but it was hardly affected in sea bass. No significant variations were recorded in the levels of protease and antiprotease. Peroxidase reached its maximum level in seabream at 02:00, the same time that it reached its minimum level in sea bass. Lysozyme showed little variation in seabream, but it was significantly lower at 14:00 than during the rest of the cycle in sea bass. Finally, different interspecific variations on bactericidal activity against Vibrio harveyi were recorded. The findings demonstrate that the immune parameters present in skin mucus of these important fish species are affected by the light–dark cycle and that there are substantial interspecies differences.