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The development of oocyte cryopreservation techniques in blue mussels Mytilus galloprovincialis

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  • Aquaculture
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Abstract

Reliable techniques for the cryopreservation of both sperm and oocytes of the blue mussel Mytilus galloprovincialis Lamarck would increase the availability of seed supplies out-of-season and enhance efficiency in selective breeding. We have investigated the optimal cryo-technique for blue mussel oocytes. The toxicity of three cryoprotective agents (CPAs) [dimethyl sulfoxide (DMSO), ethylene glycol (EG) and propylene glycol (PG)] at different concentrations (1–5 M) and exposure times (0.25–30 min) were investigated for mussel oocytes at room temperature (20 °C) or on ice. The same CPAs (1, 1.5 and 2 M) as well as three different cryoprotectant mixtures [1.5 M EG + 0.2 M trehalose + 100 % Milli-Q water (EGTM); 1.5 M EG + 0.2 M trehalose + 75 % Milli-Q water + 25 % seawater; 1.5 M EG + 0.2 M sucrose + 100 % Milli-Q water] were tested by comparing the post-thaw oocyte fertilization rate after using the slow-cooling method. Vitrification was also examined; however, this method failed to produce any post-thaw surviving oocytes. Among the tested CPAs, EG was the least toxic to oocytes. There was a tendency for the equilibration of CPAs on ice to achieve a higher oocyte fertilization rate compared with that at room temperature, and this difference was significant at concentrations of 3 and 4 M (P < 0.01). The DMSO, EG and PG treatments all resulted in post-thaw fertilization, with EGTM achieving the highest number of surviving oocytes (32 %). At the optimal seeding temperature (−7 °C), the addition of 0.2 M trehalose to EG resulted in a better fertilization rate of post-thawed oocytes than the addition of 0.2 M sucrose. All of the treatments evaluated produced D-larvae from post-thawed oocytes, although the rates were low.

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Acknowledgments

The authors would like to thank Assoc. Prof. David Stone and Dr. Ron Smernik for their assistance in proofreading this manuscript.

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Authors and Affiliations

  1. Dalian Ocean University, Dalian, China

    Hanru Wang & Meiqing Wang

  2. South Australia Research and Development Institute, West Beach, SA, 5024, Australia

    Xiaoxu Li, Steven Clarke & Mark Gluis

  3. School of Animal and Veterinary Sciences, University of Adelaide, Roseworthy Campus, Roseworthy, SA, Australia

    Hanru Wang

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  1. Hanru Wang
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  2. Xiaoxu Li
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  4. Steven Clarke
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  5. Mark Gluis
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Correspondence to Xiaoxu Li.

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Wang, H., Li, X., Wang, M. et al. The development of oocyte cryopreservation techniques in blue mussels Mytilus galloprovincialis . Fish Sci 80, 1257–1267 (2014). https://doi.org/10.1007/s12562-014-0796-9

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  • DOI: https://doi.org/10.1007/s12562-014-0796-9

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