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

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

doi:10.1016/j.aquaculture.2019.734554

Cited by 13 publications

(19 citation statements)

References 41 publications

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“…Research on other mollusc species like Pacific oyster (Crassostrea gigas) or blue mussel (Mytilus galloprovinciallis) indicate that there is not strong inter-species variability in the response to cryopreservation. In general, EG is the most used CPA for larvae cryopreservation in combination with other non-permeating CPAs like TRE or polyvinylpyrollidone (PVP) [3,6,8,9,16,19,24,25,26,28]. In this study, it was found that higher concentrations of EG, in combination with TRE at a higher concentration and PVP were generally better for survival than lower EG concentrations.…”

Section: Discussionmentioning

confidence: 81%

Development of a method to cryopreserve Greenshell mussel™ (Perna canaliculus) veliger larvae

et al. 2020

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Cryopreservation of larvae of Greenshell TM mussel Perna canaliculus, the most cultivated species in New Zealand, can provide flexibility for selective breeding programmes and enhance its global production. In this study, we set out to develop a reliable protocol for freezing D-stage larvae of Greenshell™ mussels that ensured long-term survival for successful rearing of thawed larvae in the hatchery. The effects of different combinations of cryoprotecting agents (CPA), varying CPA equilibration times, larval concentrations per straw as well as different larval development stages (48 h vs 72 h old) were evaluated by assessing the behavioural response (swimming activity, algal consumption), shell size and survival of larvae, up to 4 days post-thawing. The protocol yielding the best larval performances was a combination of the following CPA (final concentrations): 14% ethyleneglycol (EG) + 0.6M trehalose (TRE) + 1% polyvinyl-pyrrolidone (PVP), prepared with Milli-Q water. Stocking densities ranging from 50,000 to 150,000 larvae per straw (0.25 mL) and a 20 min equilibration time gave the best results, while no significant differences in fitness were found between larvae cryopreserved at 48h nor 72h-old. Using the improved cryopreservation protocol, over 50% of previously cryopreserved D-larvae were able to survive after 4 days of rearing, compared with 65% in the unfrozen control. More importantly, about one third of thawed larvae were able to swim and feed, and to potentially develop further. These findings contribute to enhance the selective breeding programmes for this species.

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Section: Discussionmentioning

confidence: 81%

Development of a method to cryopreserve Greenshell mussel™ (Perna canaliculus) veliger larvae

et al. 2020

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“…Larval cryopreservation has been widely recognized as a reliable and effective technique to conserve germplasm. This technique could be applied to the aquaculture industry to benefit the process of hatchery management, including providing a year‐round supply of larvae (Labbé et al, 2018; Liu et al, 2020). In marine bivalve species, larval cryopreservation technique is still challenging as limited spat can be produced per processing cycle, although this technique has been tried on oysters, mussels, and clams (Martínez‐Páramo et al, 2017; Simon & Yang, 2018).…”

Section: Introductionmentioning

confidence: 99%

“…Moreover, in Greenshell mussel Perna canaliculus , Paredes et al (2012) demonstrated that only 2.8% competent pediveligers were produced from post‐thaw trochophore larvae. Recently, the programmable cryopreservation technique for trochophore larvae in M. galloprovinvialis has been developed with about 20% spat achieved (Liu et al, 2020). However, the relative mortality rate of D larvae and umbo larvae produced from post‐thaw trochophore larvae is significantly higher, whereas no significant difference was found at eyed larvae and spat in comparison with fresh larvae (Liu et al, 2020), leading to reduce the efficiency on the application of larval cryopreservation technique.…”

Section: Introductionmentioning

confidence: 99%

“…Recently, the programmable cryopreservation technique for trochophore larvae in M. galloprovinvialis has been developed with about 20% spat achieved (Liu et al, 2020). However, the relative mortality rate of D larvae and umbo larvae produced from post‐thaw trochophore larvae is significantly higher, whereas no significant difference was found at eyed larvae and spat in comparison with fresh larvae (Liu et al, 2020), leading to reduce the efficiency on the application of larval cryopreservation technique. Therefore, understanding the cryodamage on larval quality is necessary, relevant, and important.…”

Section: Introductionmentioning

confidence: 99%

“…Studies on the larval cryopreservation technique in marine bivalve species have primarily focused on the protocol development/optimization stage using the percentage of D‐larvae as a quality assessment indicator (Liu et al, 2020; Martínez‐Páramo et al, 2017; Simon & Yang, 2018). There is little information on cryodamage on larval quality using other methods (Martínez‐Páramo et al, 2017; Simon & Yang, 2018; Suneja et al, 2014) except for those at the molecular and cellular levels, which have shown potentials to understand the mechanisms of cryodamage and provide useful information to improve the post‐thaw larval survival.…”

Section: Introductionmentioning

confidence: 99%

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Effects of cryopreservation on redox status and gene expression of trochophore larvae in Mytilus galloprovincialis

Liu

Catalano

Qin

et al. 2021

J World Aquaculture Soc

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The relative mortality rate of trochophore larvae exposed to cryopreservation is significantly higher at early larval developmental stages in blue mussels Mytilus galloprovincialis.However, little is known about cryodamage on the larval quality at cellular and molecular modifications in aquatic species, including M. galloprovincialis. This study aimed to investigate the effects of established cryopreservation technique on reactive oxygen species (ROS), total antioxidant capacity, apoptotic and antioxidant enzyme related gene expressions of trochophore larvae in M. galloprovincialis.The results showed that significantly lower D-larvae rate was produced from the cryopreserved trochophore larvae compared to fresh (p <.05). Nevertheless, there was no significant difference in the ROS production, total antioxidant capacity, and small molecular antioxidant capacity between fresh and cryopreserved trochophore larvae (p >.05). Similarly, the cryopreservation procedures had no adverse effects on the apoptotic and antioxidant enzyme related gene expression patterns between fresh and cryopreserved trochophore larvae (p >.05). These results indicate that the cryopreservation technique applied to M. galloprovincialis larvae in this study may not compromise the redox status of the post-thaw trochophore larvae.

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Investigation on redox status and gene expression related to larval cryopreservation in the Pacific oyster Crassostrea gigas

et al. 2022

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

Cited by 13 publications

References 41 publications

Development of a method to cryopreserve Greenshell mussel™ (Perna canaliculus) veliger larvae

Development of a method to cryopreserve Greenshell mussel™ (Perna canaliculus) veliger larvae

Effects of cryopreservation on redox status and gene expression of trochophore larvae in Mytilus galloprovincialis

Investigation on redox status and gene expression related to larval cryopreservation in the Pacific oyster Crassostrea gigas

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