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Suitable methods for cryopreservation of semen from Atlantic halibut, Hippoglossus hippoglossus L.

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Abstract

Decrease in the quality and quantity of Atlantic halibut, Hippoglossus hippoglossus L., semen towards the end of the reproductive season hampers production of good-quality embryos. Therefore, cryopreservation of spermatozoa is a method showing potential to facilitate controlled reproduction in Atlantic halibut. The present study aimed at establishing the appropriate cryopreservation procedure. We tested 20 extenders composed of four various diluents and five cryoprotectants (DMSO, DMA, methanol, propylene glycol, and glycerol) to determine the best extender. Then, we examined cryopreservation quality using various methods of loading and various volumes of cryopreserved samples. In most of the tested variants, sperm diluted with an extender showed high motility after 24-h incubation despite the high osmotic pressure of the extender. Modified turbot extender (MTE) was the best of the tested diluents, securing the highest post-thaw motility (P < 0.05), and DMSO, DMA, and methanol were the best cryoprotectants (P < 0.05). There was no significant effect of 15-min equilibration of semen in MTE-based extenders prior to freezing on post-thaw motility (P > 0.05). MTE-based extender was chosen as the most suitable. Semen cryopreserved in straws, Eppendorfs or Ziploc bags in volumes ranging from 0.25 to 20 ml showed similar high fertilization ability. Survival of larvae produced with the cryopreserved sperm did not differ from controls produced with freshly collected sperm. Motility 3 h after thawing was high but depended on the type of cryoprotectant and the volume of cryopreserved sperm (P < 0.05). The developed cryopreservation procedure has been applied at our Atlantic halibut breeding station for seed production.

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Abbreviations

DMA:

Dimethyl acetamide

DMSO:

Dimethyl sulfoxide

HBSS:

Hanks’ balanced salt solution

MTE:

Modified turbot extender

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Acknowledgments

We wish to thank Norwegian Halibut AS for providing us with halibut semen. We are much indebted to L. Schei, F. Almendingen, T. Skaalsvik, K. A. Karlsen, and B. Eggen for excellent technical assistance during the study. We thank Forskningsutvalget, Bodø University College, Norway, for financial support for the study.

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  1. Faculty of Biosciences and Aquaculture, Bodø University College, 8049, Bodø, Norway

    Igor Babiak, Sylvie Bolla & Oddvar Ottesen

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  1. Igor Babiak
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  2. Sylvie Bolla
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  3. Oddvar Ottesen
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Correspondence to Igor Babiak.

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Babiak, I., Bolla, S. & Ottesen, O. Suitable methods for cryopreservation of semen from Atlantic halibut, Hippoglossus hippoglossus L.. Aquacult Int 16, 561–572 (2008). https://doi.org/10.1007/s10499-008-9167-9

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  • DOI: https://doi.org/10.1007/s10499-008-9167-9

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