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RESEARCH ARTICLE
Contents Vol 18(3)

Development of sperm sexing and associated assisted reproductive technology for sex preselection of captive bottlenose dolphins (Tursiops truncatus)

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

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

B SeaWorld and Busch Gardens Reproductive Research Center, 500 SeaWorld Drive, San Diego, CA 92109, USA.

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

Reproduction, Fertility and Development 18(3) 319-329 https://doi.org/10.1071/RD05108
Submitted: 18 September 2005  Accepted: 1 November 2005   Published: 17 January 2006

Abstract

Research was conducted to develop sperm sorting and novel sperm preservation methodologies for sex predetermination in the bottlenose dolphin (Tursiops truncatus) using artificial insemination. In Study 1, the effect of seminal plasma (SP), sperm concentration and freezing rate (FR) on in vitro sperm quality of liquid-stored, non-sorted spermatozoa was examined. There was no effect (P > 0.05) of prefreeze SP addition on post-thaw quality (progressive motility, kinetic rating, sperm motility index (SMI), viability and acrosome integrity). Post-thaw motility parameters and viability were higher (P < 0.05) for slow FR than fast FR samples. In Study 2 investigating the effects of liquid storage and sorting on sperm quality, motility and SMI after sorting and centrifugation were lower (P < 0.05) than those of the initial ejaculate. The sort rate for enrichment (91 ± 4% purity) of X- and Y-bearing spermatozoa was 3400 ± 850 spermatozoa sex−1 s−1. In Study 3, compared with a modified straw method, directional freezing resulted in enhanced in vitro quality of sorted and non-sorted spermatozoa derived from liquid-stored semen (P < 0.05). In Study 4, endoscopic insemination of three dolphins with sorted, frozen–thawed X-bearing spermatozoa resulted in one conception and the birth of a female calf. High-purity sorting of dolphin spermatozoa, derived from liquid-stored semen, can be achieved with minimal loss of in vitro sperm quality and samples are functional in vivo.

Extra keywords: artificial insemination, cetacean, cryopreservation, sex predetermination, sperm sorting.


Acknowledgments

Animal care, animal training, curatorial and veterinary staff at SeaWorld California, USA Navy Marine Mammal Program and Dolphin Quest are thanked for assistance with animal handling and sample collection/analysis/transport, in particular Ms K. Steinman, Mr G. Biedenbach, Ms W. Coleman, Dr C. Dold, Mr L. Fish, Mr B. Hoffman, Dr E. Jensen, Dr G. Levine, Dr T. Reidarson, Mr T. Ryan, Dr T. Schmitt, Dr C. Smith, Mrs L. Smith, Dr J. Sweeney, Mrs P. Thomas, Mrs M. Tucker and Dr B. Van Bonn. Mr B. Andrews and Dr J. McBain are also thanked for invaluable support and Mr K. M. Evans for expert assistance with sorting. Dr M. Jacobson and staff at XY, Inc., Fort Collins, CO, USA and Mr J. Moreno, Mr M. Rosenstein and staff at Inguran (Navasota, TX, USA) are thanked for allowing the use of their facilities. Mrs B. Ben-Yosef and Dr A. Arav are thanked for technical assistance with the MTG-516. This project was funded by SeaWorld Corporation and is a SeaWorld Technical Contribution no. 2005-02-T.


References

Andrews B. (2000). Bottlenose dolphin management: past, present and future. In ‘The Bottlenose Dolphin Breeding Workshop’. (Eds D. Duffield and T. R. Robeck.) pp. 7–15. (AZA Marine Mammal Taxon Advisory Group: Silver Springs, MD, USA.)

Arav, A. , Yavin, S. , Zeron, Y. , Natan, D. , Dekel, I. , and Gacitua, H. (2002a). New trends in gamete’s cryopreservation. Mol. Cell. Endocrinol. 187, 77–81.
| Crossref | GoogleScholarGoogle Scholar | PubMed | Dudley M., Duffield D. A., and Shell E. D. (2004). ‘Bottlenose Dolphin North American Studbook.’ (Marine Mammal Taxon Advisory Group, American Zoo and Aquarium Association (AZA): Silver Spring, MD, USA.)

Gacitua, H. , and Arav, A. (2005). Successful pregnancies with directional freezing of large volume buck semen. Theriogenology 63, 931–938.
| Crossref | GoogleScholarGoogle Scholar | PubMed | Hermes R., Arav A., Saragusty J., Goeritz F., Pettit M., Blotter S., Flach E., Eshkar G., Boardman W., and Hildebrandt T. (2003). Cryopreservation of spermatozoa from Asian elephant (Elephas maximus) using directional freezing. Proc. Am. Assoc. Zoo Vet., 264. [Abstract]

Hollinshead, F. K. , O’Brien, J. K. , Gillan, L. , Meyers, M. , Evans, G. , and Maxwell, W. M. C. (2004a). Liquid storage of flow cytometrically sorted ram spermatozoa. Theriogenology 62, 587–605.
| Crossref | GoogleScholarGoogle Scholar | PubMed | O’Brien J. K., Crichton E. G., Evans K. M., Schenk J. L., Stojanov T., Evans G., Maxwell W. M. C., and Loskutoff N. M. (2002). Sex ratio modification using sperm sorting and assisted reproductive technology – a population management strategy. In ‘The 2nd International Symposium of Assisted Reproductive Technologies for the Conservation and Genetic Management of Wildlife, Omaha, Nebraska, 28–29 September 2002’. pp. 224–231. (Henry Doorly Zoo: Omaha, NE, USA.)

O’Brien, J. K. , Hollinshead, F. K. , Evans, K. M. , Evans, G. , and Maxwell, W. M. C. (2004). Flow cytometric sorting of frozen–thawed spermatozoa in sheep and non-human primates. Reprod. Fertil. Dev. 15, 367–375.
| Crossref | GoogleScholarGoogle Scholar | PubMed | Robeck T. R., Atkinson S., and Brook F. (2001). Reproduction. In ‘CRC Handbook in Marine Mammal Medicine’, 2nd edn. (Eds L. Dierauf and F. Gulland.) pp. 193–236. (CRC Press: Boca Raton, FL, USA.)

Robeck, T. R. , Greenwell, M. , Boehm, J. R. , Yoshioka, M. , Tobayama, T. , Steinman, K. , and Monfort, S. T. (2003). Artificial insemination using frozen–thawed semen in the Pacific white-sided dolphin (Lagenorhynchus obliquidens). Proc. Int. Assoc. Aquatic Anim. Med. 34, 50–54.
Wells R. (2000). Reproduction in wild bottlenose dolphins: overview of patterns observed during a long-term study. In ‘The Bottlenose Dolphin Breeding Workshop’. (Eds D. Duffield and T. R. Robeck.) pp. 57–73. (AZA Marine Mammal Taxon Advisory Group: Silver Springs, MD, USA.)