Abstract: Background and Objective: Clarias macrocephalus is an important aquaculture species and have generated much interest in artificial propagation, especially in enhancing the broodstock performance and larval quality for aquaculture production. This study examined the broodstock performance effects of the combined zinc amino acid and melatonin treatment to the female walking catfish, Clarias macrocephalus. Materials and Methods: The varied zinc amino acid levels and melatonin of Control (0 ppm zinc amino acid and melatonin), MZn1 (100 ppm zinc amino acid and 50 mg kg–1 melatonin) and MZn2 (200 ppm zinc amino acid and 50 mg kg–1 melatonin) were mixed in the diet comprised of isonitrogenous and isocaloric of 37% crude protein and 9.3% crude lipid and were fed to the female catfish. Zinc accumulation, maturation analysis, breeding performance and immune analysis were evaluated. Results: The results of this study revealed that the combined zinc amino acid and melatonin treatment (MZn1 and MZn2) significantly increased the maturation parameters and reproductive performance after 8 weeks of treatment. There was also a significant increase in haematological parameters within the combined zinc amino acid and melatonin treatment such as red blood cell count and haematocrit. Conclusion: This study recommended the combined zinc amino acid and melatonin treatment of the dosage of 200 ppm zinc amino acid and 50 mg kg–1 melatonin (MZn2) to enhance the female maturation of C. macrocephalus.