Tilapias are becoming increasingly popular culture fish because of their superior culture adaptability. In recent years, there has been a great interest in the use of probiotics in fish aquaculture. The objectives of the present study were to evaluate the effect of dietary graded levels (0, 5, 10, and 15 g/kg commercial diet, referred to treatments numbers T1, T2, T3, and T4, for males and T5, T6, T7, and T8 treatments for females) of a new probiotic Hydroyeast Aquaculture(®) on hematological and biochemical parameters, serum sex hormones, and the reproductive efficiency parameters of the adult Nile tilapia Oreochromis niloticus for 8 weeks. Results revealed that high levels of probiotics diet, 15 g (T4, ♂) and 10 g (T7, ♀) probiotic/kg diet, significantly (P ≤ 0.05) enhanced the physiological responses (hematological as well as serum biochemical parameters) together with, reproductive performances (sex hormones, testes and sperm quality parameters, absolute and relative fecundity, and ovarian measurements). Therefore, it could be conclude that Hydroyeast Aquaculture(®) is useful at levels of 15 g (T4) and 10 g (T7)/kg diet in improving the reproductive efficiency of adult O. niloticus males and females, respectively. Thus, the use of Hydroyeast Aquaculture(®) may be economically important for fish hatcheries.