Development of a sperm cryopreservation protocol for the mud spiny lobster, Panulirus polyphagus

Abstract

This study aimed to develop a cryopreservation protocol for spermof themud spiny lobster, Panulirus polyphagus. Spermof P. polyphaguswere successfully cryopreserved using a protocolwith cooling periods of 15min per temperature, 25, 20, 16, 4, 2, −4, −20, −80, and −150 °C, followed by immediate storage in liquid nitrogen (at −196 °C). The efficacy of the cryopreserved protocolwas determined by assessing the viability of sperm. The optimal thawing temperature for cryopreservation of sperm was 26 °C for 30 s, with a viability rate of 76.09% ± 7.81. At room temperature, −20 and −80 °C, 10% glycine provided the highest percentage of sperm viability at 91.87 ± 2.03% (5 min at room temperature), 91.31 ± 2.65% (6 h at −20 °C) and 75.88 ± 10.81% (6 h at −80 °C). In conclusion, we developed a protocol (Protocol I) for the successful cryopreservation of P. polyphagus sperm using Ca-F saline as an extender and 10% glycine as a cryoprotectant.