This study evaluated the effect of dietary aflatoxin B1 (AFB1) on growth, milt and egg quality in matured Nile tilapia (Oreochromis niloticus). Triplicate groups of Nile tilapia (initial body weight 24.1 ± 2.6 g) were fed with either of four diets (Diets 1 to 4) designed to contain 0, 20, 200 and 2000 μg AFB1 kg−1 diets for 24 weeks. After 24 weeks of AFB1 exposure, growth was significantly (P <0.05) different between the control and the AFB1 exposed treatments in both sexes. No significant differences were observed in 17β-oestradiol, absolute fecundity, oocytes volume and diameters between AFB1 exposure groups and the control group. However, we observed a significant reduction in relative fecundity and gonad somatic index (GSI) in females fed 2000 μg AFB1 kg−1 diet. On the other hand, we observed significant differences (P <0.05) in gonadosomatic index (GSI), testosterone, milt count and motility between males in the control group and AFB1 treatments. We conclude that rearing Nile tilapia with aflatoxin-contaminated diets for a prolonged period affects milt quality, fecundity (at higher doses) and growth performance. This implies that for optimal seed production, provision of aflatoxin free diets should be part of the management practices in Nile tilapia hatcheries.