Cryopreservation of strip spawned sperm using programmable freezing technique in the blue mussel Mytilus galloprovincialis

Liu, Yibing; Liu, Shiwen; Liu, Bingli; Qin, Jianguang; Xu, Tong; Li, Xiaoxu

doi:10.1007/s00343-019-7252-8

Cited by 9 publications

(8 citation statements)

References 31 publications

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“…In Chilean scallop, swollen, folded or broken plasma membranes were reported, after thawing (Espinoza et al, 2010). In blue mussel (Mytilus galloprovincialis) sperm plasma membrane integrity decreased from 90% in fresh sperm to 21_25% in thawed cells (Liu et al, 2016), suggesting a similar quality to that found in our work. Modifications of the plasma membrane may affect sperm osmoregulation capacity and its fluidity.…”

Section: Discussionsupporting

confidence: 84%

The quality of great scallop (Pecten maximus) sperm after thawing

Suquet

Gourtay

Donval

et al. 2016

General and Comparative Endocrinology

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Most publications devoted to the cryopreservation of mollusc sperm have focused on the definition of technical protocols, avoiding the description of sperm quality after thawing. The present study investigated the effects of cryopreservation on sperm quality in the great scallop. Wild scallop were fished during the natural spawning period and conditioned in the hatchery before use. Sperm samples were obtained after intragonadal injection of serotonin and cryopreserved using a previously published protocol. Sperm quality was assessed using a panel of four parameters: sperm motility characteristics, using a computer assisted sperm analysis plugin with Image J, intracellular ATP content using an ATP-Lite kit, sperm integrity, using flow cytometry and sperm morphology, using transmission electron microscopy. For each parameter, fresh (control) and thawed spermatozoa were compared. A significant decrease of both the percentage of motile spermatozoa (reduction: 75%) and sperm swimming speed (86%) were observed for thawed sperm compared with fresh sperm. The percentage of living spermatozoa, as assessed using flow cytometry, was significantly lower for thawed sperm (72.4±2.5%) compared with fresh sperm (86.4±1.1). However, no significant difference of intracellular sperm ATP content was observed between fresh and thawed sperm. Post thawing, while some spermatozoa showed little or no morphological differences compared with fresh sperm, others had undergone drastic changes, including swelling of the plasma membrane, structural alterations of the chromatin and damage to mitochondria. In conclusion, the descriptive parameters studied in the present work showed that the quality of thawed great scallop sperm was lower than that of fresh cells but was still sufficient for use in aquaculture programs and sperm cryobanking for this species.

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Section: Discussionsupporting

confidence: 84%

The quality of great scallop (Pecten maximus) sperm after thawing

Suquet

Gourtay

Donval

et al. 2016

General and Comparative Endocrinology

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“…We observed swelling in the plasma membrane after the freeze/thaw process. Similar damage was reported in the Chilean scallop Argopecten purpuratus (Espinoza et al 2010), blue mussel Mytilus galloprovincialis (Liu et al 2016) and great scallop Pecten maximus (Suquet et al 2016).…”

Section: Discussionsupporting

confidence: 75%

“…Mytilus galloprovincialis (Di Matteo et al 2009;Liu et al 2016), Argopecten purpuratus (Espinoza et al 2010), Ostrea edulis (Horváth et al 2012), Pecten maximus (Suquet et al 2016), and Pinctada angulata (Riesco et al 2017). Lyons et al (2005) and Acosta-Salmon et al (2007) reported the effect of several cryoprotectant additives on P. margaritifera sperm motility after freeze/thawing and Hui et al (2011) reported reduced viability of spermatozoa after freeze/thawing.…”

Section: Introductionmentioning

confidence: 99%

Quality assessment of cryopreserved black-lip pearl oyster Pinctada margaritifera spermatozoa

Demoy-Schneider¹,

Schmitt²,

Pennec

et al. 2018

Aquaculture

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High quality of sperm is essential to a high fertilization rate, especially post-cryopreservation. Assessment of sperm integrity, motility and energy reserves before cryopreservation is necessary for selection of milt with optimal fertilizing potential. We describe the effect of cryopreservation on the quality of black-lip pearl oyster, Pinctada margaritifera var. cumingii sperm. Evaluated quality indices of fresh and frozen/thawed P. margaritifera spermatozoa, included morphology, ultrastructure and motility characteristics relative to the energy content (ATP) and its capacity to be sustained by mitochondrial respiration. Morphology and ultrastructure were quantitatively evaluated using images obtained by optical microscopy assisted by the Image J software and TEM, respectively. Sperm motility was assessed using Image J software combined with a computer assisted sperm analysis plugin adapted for assessing P. margaritifera spermatozoa. Other sperm quality parameters evaluated included O 2 consumption, ATP content, and creatine kinase activity. Frozen/thawed spermatozoa exhibited damage to the head but retained a compact spherical shape. Sperm motility indicators showed a significant decrease in quality resulting from the freeze/thaw process. The percent of motile cells was 54% compared to 84% in fresh sperm, O 2 consumption was 4.8 compared to 44 nanomol min −1 , ATP content was 0.72 nmol/10 9 spermatozoa in the activating medium compared to 4.54 nmol/10 9 spermatozoa, and creatine kinase activity was 9.06 × 10 −5 IU mg −1 protein compared to 12.5 × 10 −5 IU mg −1 protein. The cryopreservation protocol allowed obtaining an acceptable motility rate after thawing, confirming the predictive value of sperm motility measurements before cryopreservation in terms of their ability to withstand freezing process. Highlights ► Quality of the frozen/thawed black-lip pearl oyster Pinctada margaritifera spermatozoa. ► Progress to an efficient cryoconservation method for P. margaritifera spermatozoa. ► Preservation of P. margaritifera through the durable development of the pearl oyster culture in French Polynesia.

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“…Addition of sugar supplements to the extender has shown positive effects in Greenshell mussel sperm when associated with DMSO as a permeable CPA (Smith et al 2012), while no effects were observed on blue mussel (Mytilus galloprovincialis) sperm when they were combined with the same permeable CPA (Liu et al 2015c). In the present study, no improvement in the post-thaw sperm motility was revealed by the addition of trehalose or glucose to the extender, when associated with PEG, a non-permeable CPA.…”

Section: Discussioncontrasting

confidence: 57%

Optimizing sperm cryopreservation in great scallop (Pecten maximus)

Suquet

Malo

Gourtay

et al. 2016

Aquat. Living Resour.

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A basic cryopreservation protocol for great scallop sperm was recently published, with which thawed sperm survival remained limited to 25% of control values (Suquet et al. 2014). We designed here a set of five experiments to test ways of improving thawed sperm survival and storage by studying the effects of (1) cryoprotectant concentration; (2) sugar incorporation in the extender; (3) egg yolk addition to the extender; (4) sperm dilution rate; and (5) storage capacity of thawed sperm at 4 • C. A sixth experiment was then performed to compare rearing performances of larvae produced using thawed sperm from the optimal protocol defined in the present study with those produced using fresh sperm. During the first experiment, a significantly higher percentage of motile spermatozoa was recorded after thawing when the treatment had included 15 or 20% polyethylene glycol (PEG). In the second experiment, no significant improvement in post-thaw sperm motility was recorded when trehalose or glucose was added to the extender. In the third experiment, adding 10 or 20% egg yolk to the extender significantly decreased the percentage of motile sperm. During the fourth experiment, significantly higher percentages of motile sperm were observed for dilution rates ranging from 3 : 1 to 1 : 1 than for higher dilution rates. In the fifth experiment, a significant decrease in the percentage of post-thawed sperm was observed for storage durations of 30 and 60 min, relative to shorter durations. During the last experiment, similar rearing performances were observed between larvae produced using fresh and thawed sperm, up to 9 days post fertilization. In conclusion, the high survival of sperm recorded after thawing (67% of the control level) and the high rearing performances of progenies obtained using thawed sperm validate the use of the present improved cryopreservation protocol for sperm cryobanking in great scallop.

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Cryopreservation of strip spawned sperm using programmable freezing technique in the blue mussel Mytilus galloprovincialis

Cited by 9 publications

References 31 publications

The quality of great scallop (Pecten maximus) sperm after thawing

The quality of great scallop (Pecten maximus) sperm after thawing

Quality assessment of cryopreserved black-lip pearl oyster Pinctada margaritifera spermatozoa

Optimizing sperm cryopreservation in great scallop (Pecten maximus)

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