The aim of this study was to investigate the melatonin feeding administration to the first puberty stage in male broodstock of the Walking catfish, Clarias macrocephalus. The potential of melatonin (at concentrations of 0 (control), 0.05 (Mt0.05) and 0.25 (Mt0.25) mg/kg) to enhance male reproductive system (gonad histology, gonadosomatic index and sperm abnormality) was investigated. In addition, the melatonin feeding enhancement of C. macrocephalus gonadosomatic index and lower sperm abnormality were found consistent with higher spermatozoa cells in melatonin-treated (Control: 38.8%, Mt0.05: 48.6%, and Mt0.25: 85.4%) fishes where the melatonin feeding treatment increased the gonadosomatic index (0.43 – 0.68%; p: 0.001) and decreased the sperm abnormality (7.7 – 130.8%; p: 0.001) after eight weeks of treatment. The present results supported the concept of melatonin being an important hormone in regulating the reproductive system for male catfish. Thus, melatonin feeding administration helps in enhancing the first puberty stage of male C. Macrocephalus.