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RESEARCH ARTICLE
Contents Vol 20(7)

Semen collection, characterisation and artificial insemination in the beluga (Delphinapterus leucas) using liquid-stored spermatozoa

J. K. O’Brien A B D , K. J. Steinman A , T. Schmitt C and T. R. Robeck A
+ Author Affiliations
- Author Affiliations

A SeaWorld and Busch Gardens Reproductive Research Center, 2595 Ingraham St, San Diego, CA 92109, USA.

B Centre for Advanced Technologies in Animal Genetics and Reproduction, Faculty of Veterinary Science, University of Sydney, NSW 2006, Australia.

C SeaWorld California, 500 SeaWorld Dr, San Diego, CA 92109, USA.

D Corresponding author. Email: justineo@vetsci.usyd.edu.au

Reproduction, Fertility and Development 20(7) 770-783 https://doi.org/10.1071/RD08031
Submitted: 17 February 2008  Accepted: 4 June 2008   Published: 1 August 2008

Abstract

Ejaculates were collected from a beluga (Delphinapterus leucas) to gain an understanding of sperm biology and develop a short-term sperm preservation method for use in artificial insemination (AI). Ejaculate parameters and biochemistry, semen production and serum testosterone concentrations of an adult male were characterised for 21 months. Sperm viability, acrosome integrity and morphology did not change (P > 0.05) but ejaculate volume, sperm concentration and total spermatozoa per ejaculate were higher (P < 0.05) from January to June than from July to December. Peak testosterone concentrations (P < 0.05) were observed from October to April (8.0 ± 1.6 ng mL–1). The effects of hyaluronic acid (HA), antioxidants, storage temperature and time on in vitro sperm characteristics were examined. Motility parameters and viability were improved (P < 0.05) when semen was stored at 5°C compared with 21°C. During the first 24 h of storage sperm agglutination was absent only at 5°C in the presence of HA. A nulliparous 28-year-old female was inseminated endoscopically with liquid-stored semen. A pregnancy and birth of a calf was achieved following AI for the first time in this species, thereby validating both the AI technique and the fertility of beluga spermatozoa after chilled storage in a specialised diluent.

Additional keywords: cetacean, hyaluronan, intrauterine insemination, semen storage.


Acknowledgements

Animal care, animal training, curatorial and veterinary staff at SeaWorld California and Texas are thanked for assistance with animal handling and sample collection. Mrs Mitzi Synnott and Mrs Nicole Grovhoug are particularly thanked for the initial training which led to successful collection of beluga semen, and we are grateful to all Wild Arctic Staff for ongoing collections and assistance with AI procedures. Animal Care Laboratory staff (SeaWorld California) including Ms Whitney Coleman, Mrs Leigh Ann Smith, Mrs Pam Thomas and Mrs Melinda Tucker are thanked for technical support, as is Ms Sherry Dickerson, Dr Steve Osborn and Mr Chris White (SeaWorld Texas) for sample collection and preparation. Dr Robert Anderson (Obstetrical and Gynaecology Research, Rush Medical Center, Chicago) performed the seminal plasma fructose analyses, for which we are grateful. Mr Brad Andrews is also thanked for invaluable support of this research. This project was funded by SeaWorld Corporation and is a SeaWorld Technical Contribution no. 2008–01-T.


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