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

• The development of cryopreservation protocol of Panulirus polyphagus's sperm.
• Relation of different cryoprotectants and the sperm viability.
• Producing high sperm viability from the sperm cryopreservation.

This study aimed to develop a cryopreservation protocol for sperm of the mud spiny lobster, Panulirus polyphagus. Sperm of P. polyphagus were successfully cryopreserved using a protocol with cooling periods of 15 min 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 protocol was 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.Statement of relevanceThis article is suitable with Aquaculture Journal because the findings will give a big contribution in mud spiny lobster aquaculture. Mud spiny lobster seedling especially P. polyphagus is getting less landing every year, new approach needed in ensuring enough supplies of P. polyphagus to be available. The content in this article would help developing the research innovation for the hatchery production of P. polyphagus to increase.