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Cryobiology. 2014 Apr;68(2):185-93. doi: 10.1016/j.cryobiol.2014.01.002. Epub 2014 Jan 17.

Cryopreservation of sperm in farmed Australian greenlip abalone Haliotis laevigata.

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  • 1School of Biological Sciences, The Flinders University of South Australia, Adelaide, South Australia 5042, Australia.
  • 2Dalian Ocean University, Dalian 116023, China.
  • 3School of Biological Sciences, The Flinders University of South Australia, Adelaide, South Australia 5042, Australia; Nofima, PO Box 210, Ås, Norway.
  • 4South Australian Research and Development Institute, and Marine Innovation Southern Australia, West Beach, South Australia 5024, Australia. Electronic address:


This study investigated factors important to the development of the liquid nitrogen (LN) vapor sperm cryopreservation technique in farmed greenlip abalone Haliotis laevigata, including (1) cryoprotectant agent (CPA) toxicity; (2) cooling temperature (height above LN surface); (3) thawing temperature; (4) sperm to egg ratio; and (5) sugar supplementation, using sperm motility, fertilization rate or integrity/potential of sperm components and organelles as quality assessment indicators. Results suggested that among the single CPAs evaluated 6% dimethyl sulfoxide (Me2SO) would be the most suitable for sperm cryopreservation in this species. The highest post-thaw sperm motility was achieved with the sperm that had been exposed to LN vapor for 10min at 5.2cm above the LN surface, thawed and recovered in 60 and 18°C seawater bathes, respectively after at least 2h storage in LN. The highest fertilization rates were achieved at a sperm to egg ratio of 10,000:1 or 15,000:1. Addition of 1% glucose or 2% sucrose produced significantly higher post-thaw sperm motility than 6% Me2SO alone. Among the three cryoprotectant solutions further trialled, 6% Me2SO+1% glucose produced the highest fertilization rate of 83.6±3.7%. Evaluation of sperm has shown that the addition of glucose could significantly improve the sperm plasma membrane integrity and mitochondrial membrane potential. These results demonstrated a positive role of glucose in the improvement of sperm cryopreservation in farmed greenlip abalone.

Copyright © 2014 Elsevier Inc. All rights reserved.


Farmed greenlip abalone; Glucose; Haliotis laevigata; Sperm cryopreservation

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