Effects of larval cryopreservation on subsequent development of the blue mussels, Mytilus galloprovincialis Lamarck

Abstract

In this study, the e¡ects of three commonly used chemicals, dimethyl sulphoxide (DMSO), ethylene glycol (EG), propylene glycol (PG) and their combinations with trehalose, were evaluated on the cryopreservation of Dlarvae of the blue mussel Mytilus galloprovincialis. The larvae were harvested 30 h post-fertilization at 21 1C and cryopreserved using a standard protocol in 5%, 10% or15% of DSMO, EG and PG either as single chemical solutions or in combination with 0.2 M trehalose. Among these cryoprotectants, 5% DMSO resulted in the highest post-thaw survival rate of 55.3 AE 7.8%, although it did not signi¢cantly di¡er from those with 10% and 15% EG. The addition of 0.2 M trehalose did not improve the post-thaw larval survival rates in all the combinations assessed. The cryo-e¡ects on subsequent development were evaluated using the D-larvae frozen with 5% DMSO. The results showed that cryopreservation a¡ected both larval survival and growth in this species. The relative daily mortality rate was sig-ni¢cantly higher in treated than control groups over the period from 3 h post-thaw to day 11 post-fertilization. On day 6 post-fertilization, the average larval length in the treated group was signi¢cantly smaller than that in the control. From day 11 post-fertilization, and onwards, di¡erences in these two traits were not signi¢cant between treated and control groups. On day 21 post-fertilization, about 80% of the larvae in both treated and control groups developed eyes and the normalized survival rate in the treated group was12.5%.

Survival, Growth and Reproduction of Cryopreserved Larvae from a Marine Invertebrate, the Pacific Oyster (Crassostrea gigas)

“…In blue mussel ( Mytilus galloprovincialis ), the survival of 21-day old larvae after thawing was 12.5% of the control group [5]. Only 2.8% of the frozen/thawed Greenshell mussel ( Perna canaliculus ) trochophores survived up to competent pediveligers and their size was lower than that observed on unfrozen larvae [6].…”

Exploring the evolution of marine invertebrate cryopreservation – Landmarks, state of the art and future lines of research

“…Cryopreservation of embryos and larvae as an alternative to oocyte cryopreservation have given more promising results (e.g. Asahina and Takahashi 1979, Olive and Wang 1997, Toledo and Kurorura 1990, Grout and McFadzen 1991, Lin et al 1999, Wang et al 2011, Paredes and Bellas 2014 but also new challenges. Both oocytes and embryos show high sensitivity to sub-zero temperatures, sensitivity to CPA exposure and lower permeability, embryos and larvae present some degree of organ cryoinjury or deformation of hard surfaces such as shells.…”

Cryobanking of aquatic species

“…Adding sugars (glucose or sucrose) improved the post-thaw survival of Pacific oyster (Renard, 1991) and of pearl oyster (Choi and Chang, 2003) by reducing the toxicity of cryoprotectants and decreasing injuries of thawed embryos. However, adding trehalose did not improve the post-thaw survival of blue mussel embryos (Wang et al, 2011). Regarding embryo development stage, the consensus view is that the trochophore stage is best adapted to cryopreservation in Pacific oyster (Gwo, 1995; Usuki et al, 2002), but not in pearl oyster, the best survival being observed at the D-larval stage (Choi and Chang, 2003).…”