Advertisement

Cryopreservation of Semen from Domestic Livestock

  • Harald SiemeEmail author
  • Harriëtte Oldenhof
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1257)

Abstract

In modern livestock breeding, cryopreserved semen is routinely used for artificial insemination. Sperm cryopreservation secures future reproduction, and insemination doses can be easily shipped. Processing of semen for cryopreservation can be done with minimal efforts and relatively low costs. In this chapter we describe the entire cryopreservation process for stallion and bull sperm including dilution of sperm in primary and freezing extender, cooling and packaging in straws, freezing in liquid nitrogen vapor, cryogenic storage, and thawing. Special emphasis is given on preparation of commonly used primary and freezing extenders (skim milk extender “INRA-82”, TRIS-egg yolk extender “TEY”) used in a two-step dilution approach. Furthermore the different cooling rates needed in different temperature regimes during the freezing process are being described. Cryopreservation procedures are described in case of using both specialized automated equipment and simple equipment.

Key words

Stallion Bull Semen Spermatozoa Cryopreservation 

Notes

Acknowledgments

Employees of the National Stud of Lower Saxony (Celle, Germany) and Masterrind GmbH (Verden, Germany) are acknowledged for sharing protocols.

References

  1. 1.
    Johnson MD, Cooper AR, Jungheim ES, Lanzendorf SE, Odem RR, Ratts VS (2013) Sperm banking for fertility preservation: a 20-year experience. Eur J Obstet Gynecol Reprod Biol 170:177–182CrossRefGoogle Scholar
  2. 2.
    Curry MR (2000) Cryopreservation of semen from domestic livestock. Rev Reprod 5:46–52CrossRefGoogle Scholar
  3. 3.
    Rodriguez-Martinez RM (2013) Sperm biotechnologies in domestic species: state of the art. Anim Reprod 10:268–276CrossRefGoogle Scholar
  4. 4.
    Baracaldo MI, Barth AD, Bertrand W (2006) Steps for freezing bovine semen: from semen collection to the liquid nitrogen tank. In: IVIS Reviews in veterinary medicine, International Veterinary Information Service—IVIS (eds) Ithaca, NY. www.ivis.org
  5. 5.
    Vishvanath R, Shannon P (2000) Storage of bovine semen in liquid and frozen state. Anim Reprod Sci 62:23–53CrossRefGoogle Scholar
  6. 6.
    Samper JC, Morris CA (1998) Current methods for stallion semen cryopreservation: a survey. Theriogenology 49:895–903CrossRefGoogle Scholar
  7. 7.
    Vidament M (2005) French field results (1985–2005) on factors affecting fertility of frozen stallion semen. Anim Reprod Sci 89:115–136CrossRefGoogle Scholar
  8. 8.
    Johnson LA, Weitze KF, Fiser P, Maxwell WMC (2000) Storage of boar semen. Anim Reprod Sci 62:143–172CrossRefGoogle Scholar
  9. 9.
    Didion BA, Braun GD, Duggan MV (2013) Field fertility of frozen boar semen: a retrospective report comprising over 2600 AI services spanning a four year period. Anim Reprod Sci 137:189–196CrossRefGoogle Scholar
  10. 10.
    Sieme H (2011) Freezing semen. In: McKinnon AO, Squires EL, Vaala WE, Varner DD (eds) Equine reproduction, 2nd edn. Blackwell Publishing Ltd., Chichester, UK, pp 2972–2982Google Scholar
  11. 11.
    Sieme H (2011) Semen extenders for frozen semen. In: McKinnon AO, Squires EL, Vaala WE, Varner DD (eds) Equine reproduction, 2nd edn. Blackwell Publishing Ltd., Chichester, UK, pp 2964–2971Google Scholar
  12. 12.
    Hancock JL (1957) The morphology of boar spermatozoa. J Roy Microsc Soc 76:84–97CrossRefGoogle Scholar
  13. 13.
    Thibier M, Guerin B (2000) Hygienic aspects of storage and use of semen for artificial insemination. Anim Reprod Sci 62:233–251CrossRefGoogle Scholar
  14. 14.
    Hurtgen JP (2000) Semen collection in stallions. In: Samper JC (ed) Equine breeding management and artificial insemination. W.B. Saunders Company, Pennsylvania, pp 61–69Google Scholar
  15. 15.
    Moore AI, Squires EL, Graham JK (2005) Effect of seminal plasma on the cryopreservation of equine spermatozoa. Theriogenology 63:2372–2381CrossRefGoogle Scholar
  16. 16.
    Saragusty J, Gacitua H, Rozenboim I, Arav A (2009) Protective effects of iodixanol during bovine sperm cryopreservation. Theriogenology 71:1425–1432CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Clinic for Horses-Unit for Reproductive MedicineUniversity of Veterinary Medicine HannoverHannoverGermany

Personalised recommendations