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LIU Yibing, LIU Shiwen, LIU Bingli, QIN Jianguang, XU Tong, LI Xiaoxu. Cryopreservation of strip spawned sperm using programmable freezing technique in the blue mussel Mytilus galloprovincialis[J]. HaiyangYuHuZhao, 2018, 36(6): 2351-2357

Cryopreservation of strip spawned sperm using programmable freezing technique in the blue mussel Mytilus galloprovincialis

LIU Yibing1,2, LIU Shiwen3, LIU Bingli3, QIN Jianguang2, XU Tong3, LI Xiaoxu4
1 Liaoning Ocean and Fisheries Science Research Institute, Dalian 116023, China;
2 College of Science and Engineering, The Flinders University of South Australia, Adelaide, South Australia 5042, Australia;
3 College of Fisheries and Life Science, Dalian Ocean University, Dalian 116023, China;
4 South Australian Research and Development Institute, West Beach, South Australia 5024, Australia
In this study, a programmable freezing technique has been developed for strip spawned sperm in the blue mussel, Mytilus galloprovincialis. The optimized key parameters include cooling rate, endpoint temperature, thawing temperature, sugar addition and sperm to oocyte ratio. The sperm quality was assessed by the fertilization rate or the integrity of sperm component and organelle. The highest post-thaw sperm fertilization rate was 91%, which was produced with sperm cryopreserved in 8% dimethyl sulfoxide at the cooling rate of -4℃/min from 2℃ to -30℃ before being plunged into liquid nitrogen for at least 12 h, thawed in a 20℃ seawater bath and fertilized at sperm to egg ratio of 50 000:1. The addition of glucose, sucrose or trehalose to 8% dimethyl sulfoxide could not further improve fertilization rates. The fluorescent assessments showed that the post-thaw sperm plasma membrane integrity and acrosome integrity were significantly damaged in comparison with fresh sperm.
Key words:    blue mussel|Mytilus galloprovincialis|strip spawning|sperm cryopreservation   
Received: 2017-09-04   Revised:
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