Laboratorio de Ecoloxía Costeira (ECOCOST), Departamento de Ecoloxía e Bioloxía Animal, Centro de Investigación Mariña, Universidade de Vigo, Vigo, Spain.
Sci Rep. 2022 Aug 10;12(1):13632. doi: 10.1038/s41598-022-17935-0.
Shellfish aquaculture needs the development of new tools for the improvement of good practices avoiding the reliance on natural spat collection to increase production efficiently. The aim of this work was to improve the cryopreservation protocol for Mytilus galloprovincialis larvae described in Paredes et al. (in: Wolkers, Oldenhof (eds) Cryopreservation and freeze-drying protocol, methods in molecular biology, Humana Press, 2021, pp 2180, https://doi.org/10.1007/978-1-0716-0783-1_18 ). Moreover, the capability of producing adult mussels from cryopreserved 72 h-old D-larvae and potential long-term effects of cryopreservation through progenies were evaluated. The selection of 72-h old D-larvae for cryopreservation yielded 75% of recovery, higher than 50% from trochophores. The best combination was 10% Ethylene-Glycol + 0.4 M Trehalose in Filtered Sea Water (FSW) with cooling at - 1 °C/min and a water bath at 35 °C for thawing. Sucrose (SUC) solutions did not improve larval recovery (p > 0.05). At settlement, 5.26% of cryopreserved F1 larvae survived and over 70% settled. F2 cryopreservation produced 0.15% survival of spat and settlement varied from 35 to 50%. The delay of shell size showed on cryopreserved larvae declined throughout larval rearing without significant differences with controls from settlement point (p > 0.05). Long-term experiments showed that it is possible to obtain adult mussels from cryopreserved larvae and this tool does not compromise the quality of following progenies, neither for cryopreservation nor post-thawing development of them.
贝类水产养殖需要开发新工具来改进良好做法，避免依赖自然苗种采集来提高产量。本工作旨在改进 Paredes 等人描述的贻贝幼虫的冷冻保存方案（in: Wolkers, Oldenhof (eds) Cryopreservation and freeze-drying protocol, methods in molecular biology, Humana Press, 2021, pp 2180, https://doi.org/10.1007/978-1-0716-0783-1_18 ）。此外，评估了从冷冻保存的 72 小时龄 D 幼虫生产成体贻贝的能力以及冷冻保存通过后代产生的潜在长期影响。选择 72 小时龄的 D 幼虫进行冷冻保存，回收率为 75%，高于从担轮幼虫的 50%。最佳组合是在过滤海水中（FSW）中使用 10%的乙二醇+0.4M海藻糖，冷却速度为-1°C/min，解冻时水温为 35°C。蔗糖（SUC）溶液不能提高幼虫的回收率（p>0.05）。在附着时，冷冻保存的 F1 幼虫中有 5.26%存活，超过 70%的幼虫附着。F2 冷冻保存的幼虫仅有 0.15%的存活，附着率从 35%到 50%不等。冷冻保存幼虫的壳尺寸延迟在整个幼虫养殖过程中逐渐减少，与从附着点开始的对照组相比没有显著差异（p>0.05）。长期实验表明，从冷冻保存的幼虫中获得成体贻贝是可能的，而且这种工具不会影响后续幼虫的质量，无论是冷冻保存还是解冻后发育。
