Development of a programmable freezing technique on larval cryopreservation in Mytilus galloprovincialis

Yibing Liu, Mark Gluis, Penny Miller-Ezzy, Jianguang Qin, Jiabo Han, Xin Zhan, Xiaoxu Li

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)


This study investigated the factors important to the development of a programmable larval cryopreservation technique in Mytilus galloprovinvialis, including (Bayne et al., 1978) larval developmental stages (Beiras et al., 2003); cryoprotectant agents (CPAs) (Castillo-Martín et al., 2013); thawing temperatures (Chao et al., 1997); sucrose concentrations to remove CPA after thawing and (Choi and Chang, 2003) straw volumes, using D-larval rate as the post-thaw survival indicator. The results showed that larvae at 25 h post-fertilization (PF) had the highest resistance to cryopreservation. A post-thaw D-larval rate higher than 80% was achieved when the larvae were cryopreserved with 10% ethylene glycol +7.5% Ficoll +0.2% polyvinylpyrrolidone, thawed at 28 °C and used 9% sucrose solution as the medium to remove CPA after thawing. No significant difference in post-thaw D-larval rates (P > 0.05) was found between larvae cryopreserved in 0.25 mL and 0.5 mL straws. The performance comparison experiment showed that although a significantly lower survival rate (P < 0.05) was found in cryopreserved larvae than that in fresh larvae at day 2 PF (D larvae), no significant difference (P > 0.05) was shown on relative mortality rate from day 8 PF (early umbonal larvae) to day 32 PF (spat) and on shell length at day 8 PF and day 32 PF between fresh and cryopreserved larvae. Therefore, the larval cryopreservation technique developed in this study would enhance the breeding program and all year-round hatchery production in this species.

Original languageEnglish
Article number734554
Number of pages6
Publication statusPublished - 1 Feb 2020


  • Ficoll
  • Larval cryopreservation
  • Mytilus galloprovinvialis
  • Polyvinylpyrrolidone


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