This study evaluated the effects of feeding time strategies on growth performance, hematological parameters, physiological responses, and organ histology in African catfish (Clarias gariepinus) juveniles. A total of 130 fish (6.03 ± 0.16 g) were randomly assigned to four feeding regimes for eight weeks in triplicate. The control group (A) was fed three times daily (07:00, 12:00, and 06:00 h), while Groups B, C, and D were fed twice daily at different intervals: morning–afternoon (07:00 and 12:00 h), afternoon–evening (12:00 and 06:00 h), and morning–evening (07:00 and 06:00 h), respectively. Results showed that final weight, specific growth rate, weight gain, feed intake, protein efficiency ratio, protein productive value, and protein growth rate were significantly reduced in Group B and moderately lower in Group C, while the feed conversion ratio exhibited an opposite trend compared to the control. Furthermore, fish in Group B recorded significantly higher alanine aminotransferase, lactate dehydrogenase, and urea levels, whereas total protein, glucose, total cholesterol, and high-density lipoprotein were markedly reduced. Similarly, white blood cell, red blood cell, and hemoglobin concentrations were lower in Groups B and C than in the control and Group D. Group B also exhibited the lowest hematocrit value, while Groups C and D showed comparable values to the control. Conversely, mean corpuscular hemoglobin was significantly higher in Group B, whereas mean corpuscular hemoglobin concentration showed a reverse trend, with no significant difference in Group C relative to the control. Histological examination revealed normal hepatic architecture across all groups, with slight vacuolation in Group B, while intestinal morphology showed greater villus height, width, and muscularis thickness in Group A compared to Group B, and no marked differences between Groups C and D. Overall, the findings suggest that feeding twice daily is effective when feeding times align with the fish’s metabolic rhythm, thereby enhancing performance and physiological balance.