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Auteur Ramos-Judez, S.; Gonzalez, W.; Dutto, G.; Mylona, C.C.; Fauvel, C.; Duncan, N.
Titre Gamete quality and management for in vitro fertilisation in meagre (Argyrosomus regius) Type Article scientifique
Année 2019 Publication Revue Abrégée Aquaculture
Volume 509 Numéro Pages 227-235
Mots-Clés Argyrosomus regius; Artificial fertilisation; artificial fertilization; cultured fish; Gamete management; GnRHa; hatching rates; injections; Meagre; ovulation; Reproduction; sea-bass; sperm activation; spermatozoaoocyte ratio; time; water volume
Résumé The aquaculture of meagre (Argyrosomus regius) requires methods for the control of reproduction that enable the production of families from specific individuals for selective breeding programs. We experimentally determined the parameters required for an in vitro fertilisation protocol. A total of 14 females and 5 males (mean +/- S.D. weights of 20.45 +/- 6.22 and 15.94 +/- 2.75 kg, respectively) were used. Selected females had vitellogenic oocytes > 550 pm in diameter and males had fluid sperm upon application of abdominal pressure. Both sexes were treated with an injection of 15 mu g kg(-1) of gonadotropin-releasing hormone agonist (GnRHa) to induce oocyte maturation/ovulation and enhance sperm production. To determine the timing of ovulation and window of high egg viability, females were stripped serially every 2.5 h beginning 35 h after GnRHa treatment. Sperm was obtained 24 h after GnRHa treatment and was diluted 1/4 in modified Leibovitz for storage at 4 degrees C until use. Sperm quality parameters such as percentage initial spermatozoa motility, duration of motility, velocity and density were determined using computer assisted sperm analysis (CASA). In vitro inseminations were made in duplicate or triplicate batches of eggs from each spawn by mixing 0.5-1 mL of eggs, 20-40 mu L diluted sperm (pooled from two males) and 100 mL of seawater. Fertilisation success was examined at spermatozoa (spz): egg ratios between similar to 2000 and 400,000 spz egg(-1). The optimal time for stripping ovulated females was <= 3 h after ovulation, which was the window of optimal egg viability. Ovulation under the conditions of this study was close to 38 h after GnRHa treatment, with a range from 35 to 41 h. Beginning from 3 h after ovulation, egg viability declined probably due to overripening. Sperm diluted in Leibovitz maintained motility and velocity for as long as 7 h after collection. Spermatozoa motility (%) and average path velocity (VAP, mu m/s) of sperm samples obtained from males before GnRHa injection declined rapidly after activation compared to the samples obtained 24 h post-injection, with significant decreases respectively after 75 and 45 s. A minimum ratio of 150,000 spermatozoa egg(-1) was necessary to ensure high fertilisation success. The acquired knowledge of the present study will aid the aquaculture industry and future research on selective breeding programs for meagre.
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Numéro d'Appel MARBEC @ isabelle.vidal-ayouba @ collection 2610
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Auteur Suquet, M.; Gourtay, C.; Donval, A.; Le Goic, N.; Quere, C.; Malo, F.; Le Grand, J.; Ratiskol, D.; Mingant, C.; Fauvel, C.
Titre The quality of great scallop (Pecten maximus) sperm after thawing Type Article scientifique
Année 2016 Publication Revue Abrégée Gen. Comp. Endocrinol.
Volume 229 Numéro Pages 127-131
Mots-Clés aquaculture; crassostrea-gigas; Cryopreservation; cryopreserved spermatozoa; fertility; fertilization; fucata-martensii; Gamete; japanese pearl oyster; motility; ostrea-edulis; pacific oyster; Pecten maximus; reproduction; Sperm
Résumé 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. (c) 2016 Elsevier Inc. All rights reserved.
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ISSN 0016-6480 ISBN Médium
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Numéro d'Appel MARBEC @ alain.herve @ collection 1657
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Auteur Suquet, M.; Malo, F.; Queau, I.; Ratiskol, D.; Quere, C.; Le Grand, J.; Fauvel, C.
Titre Seasonal variation of sperm quality in Pacific oyster (Crassostrea gigas) Type Article scientifique
Année 2016 Publication Revue Abrégée Aquaculture
Volume 464 Numéro Pages 638-641
Mots-Clés age; Crassostrea gigas; cryopreservation; fish; gamete quality; Motility; Season; spawning season; Sperm ageing; Sperm quality; time
Résumé Seasonal changes of sperm quality which can affect sperm biological parameters throughout the breeding period, have been little studied in mollusc species. Controlling gamete quality would aid the management of gametes in hatcheries and the development of selection programs. The aim of the present study was to describe the changes in sperm quality of wild Pacific oysters through the spawning season by comparing sperm parameters al the beginning (May), middle (July) and end (October) of this period using a panel of bio-descriptors. These parameters were studied over the 2014 breeding season based on shed sperm collected after serotonin injection of wild breeders. A significantly higher percentage of motile sperm was observed al the end of the spawning season (+78% relative to the value observed al the beginning) althought a lower total number of spermatozoa was collected (-59%). The mean condition index of oysters, however, was no different between the three sampling dates. For intratesticular sperm, the increase of the percentage of motile sperm and Velocity of the Average Path (VAP) in relation to time post activation was not significantly different among sperm sampling periods, suggesting that the kinetic of the sperm 'maturation process was similar. Furthermore, the mean VAP observed on shed sperm did not change through the spawning season. The sub-continuous gametogenesis of Pacific oyster can help to explain why only limited consequences of sperm ageing are observed in this species. Furthermore, the effects of sperm ageing may depend on the annual reproductive pattern of Pacific oyster. Statement of relevance: This study showed the effect of sperm ageing on sperm quality parameters. This knowledge is useful for aquaculture and would support the recent trends of mollusc farming, allowing a better management of the gametes in hatchery. (C) 2016 Elsevier B.V. All rights reserved.
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Editeur Lieu de Publication Éditeur
Langue English Langue du Résumé Titre Original
Éditeur de collection Titre de collection Titre de collection Abrégé
Volume de collection Numéro de collection Edition
ISSN 0044-8486 ISBN Médium
Région Expédition Conférence
Notes Approuvé pas de
Numéro d'Appel MARBEC @ alain.herve @ collection 1687
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