Six different hatching treatments and one control were used to eliminate egg stickiness of fertilized bullseye puffer fish Sphoeroides annulatus eggs in this experiment. Each treatment was carried out in triplicate and the eggs were incubated in fiberglass tanks. A fertilization rate of 96.2% ± 2.4% was obtained and significant differences were observed between treatments. The highest hatching rate occurred at 96.4 ± 2% with the protease enzyme solution whilst the lowest rate was observed in the control (18.8 ± 5.3%). The hatching success of the bullseye puffer fish is significantly improved by eliminating egg adhesion.