To our knowledge, this is the first observation showing that fish spermatozoa (devoid of acrosome in carp) can be revived for a second motility round and that such spermatozoa are also able to fully fertilize eggs, meaning that fish spermatozoa are not dead at the end of their first motility period. Firstly activated sperm showed significantly higher percentage of motility and sperm velocity compared to secondly activated sperm. Sperm incubation in immobilizing solution improved the structural damages in activated sperm.