Register      Login
Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
Shopping Cart: ( empty )
You are here: Home > Journals > RD > RD06078
RESEARCH ARTICLE
Contents Vol 19(5)

Comparison of different sperm cryopreservation procedures on post-thaw quality and heterologous in vitro fertilisation success in the ocelot (Leopardus pardalis)

Monica A. Stoops A D , Jennifer B. Bond A , Helen L. Bateman A , Mark K. Campbell A , Gregory P. Levens B , Todd R. Bowsher B , Shannon T. Ferrell C and William F. Swanson A
+ Author Affiliations
- Author Affiliations

A Center for Conservation and Research of Endangered Wildlife, Cincinnati Zoo and Botanical Garden, 3400 Vine Street, Cincinnati, OH 45220, USA.

B Dallas Zoo, 650 South Thorton Freeway, Dallas, TX 75203, USA.

C Fort Worth Zoo, 1989 Colonial Parkway, Fort Worth, TX 76110, USA.

D Corresponding author. Email: monica.stoops@cincinnatizoo.org

Reproduction, Fertility and Development 19(5) 685-694 https://doi.org/10.1071/RD06078
Submitted: 24 July 2006  Accepted: 30 April 2007   Published: 2 July 2007

Abstract

Cryopreservation of spermatozoa from free-living ocelots (Leopardus pardalis) could benefit their conservation by facilitating gene flow between in situ and ex situ populations without requiring removal of additional cats from the wild. The objective of this study was to investigate three different methods of ocelot sperm cryopreservation to identify the most appropriate technique for use in a field environment. Male ocelots (n = 10), housed in North American zoos, were anaesthetised with tiletamine–zolazepam (7mg kg–1 bodyweight; i.m.) and subjected to a regimented electroejaculation procedure. Recovered semen was evaluated for sperm concentration, motility and morphology and processed for cryopreservation by three methods: (1) pelleting on dry ice, (2) freezing in straws over liquid nitrogen vapour; and (3) freezing in straws in a dry shipper. Frozen samples were thawed and assessed for post-thaw acrosome status, viability, motility over time and ability to fertilise viable domestic cat oocytes. Although several post-thaw sperm parameters varied (P < 0.05) among freezing methods, frozen–thawed ocelot spermatozoa from all treatments showed a similar (P > 0.05) capacity to bind, penetrate and fertilise viable domestic cat oocytes. These findings suggest that spermatozoa collected from male ocelots under field conditions may be frozen in straws either using liquid nitrogen alone or in a charged dry shipper to retain adequate functional competence after thawing for use with assisted reproductive procedures.


Acknowledgements

The authors thank the curatorial, veterinary and keeper staff at each of the collaborating institutions (African Wildlife Safari, Carnivore Preservation Trust, Cincinnati Zoo & Botanical Garden, Dallas Zoo, Fort Worth Zoo, Oglebay’s Good Zoo, Nashville Zoo) for providing access to ocelots and assistance with anaesthesia and research procedures. Special thanks are extended to Nicole Siegel, D.V.M. for assistance with the project. We also acknowledge the National Hormone and Pituitary Program of the National Institutes of Health for the donation of ovine LH and FSH. Funding for this study was provided, in part, by a grant from the National Institutes of Health (RR001588).


References

Barone, M. A. , Roelke, M. E. , Howard, J. G. , Brown, J. L. , Anderson, A. E. , and Wildt, D. E. (1994). Reproductive characteristics of male Florida panthers: comparative studies from Florida, Texas, Colorado, Latin America, and North American zoos. J. Mammal. 75, 150–162.
| Crossref | GoogleScholarGoogle Scholar | Bragin N. (2003). ‘North American Regional Studbook for the Ocelot (Leopardus pardalis).’ (Denver Zoological Gardens: Denver, CO.)

Crosier, A. E. , Pukazhenthi, B. S. , Henghali, J. N. , Howard, J. G. , Dickman, A. J. , Marker, L. , and Wildt, D. E. (2006). Cryopreservation of spermatozoa from wild-born Namibian cheetahs (Acinonyx jubatus) and influence of glycerol on cryosurvival. Cryobiology 52, 169–181.
| Crossref | GoogleScholarGoogle Scholar | PubMed | Howard J. G. (1993). Semen collection and analysis in carnivores. In ‘Zoo and Wild Animal Medicine: Current Therapy 3.’ (Ed. M. E. Fowler.) pp. 390–399. (WB Saunders: Philadelphia, PA.)

Howard J. G., Bush M., and Wildt D. E. (1986). Semen collection, analysis and cryopreservation in nondomestic mammals. In ‘Current Therapy in Theriogenology’. (Ed. D. A. Morrow.) pp. 1047–1053. (WB Saunders: Philadelphia, PA.)

Howard, J. G. , Roth, T. L. , Byers, A. P. , Swanson, W. F. , Buff, J. L. , Bush, M. , Grisham, J. , Marker-Kraus, L. , and Wildt, D. E. (1997). Successful intercontinental genome resource banking and artificial insemination with cryopreserved semen in cheetahs. J. Androl. 55((Suppl.)), 123.
International Union for Conservation of Nature and Natural Resources (2003). ‘2003 IUCN Red List of Threatened Species.’ (IUCN: Gland, Switzerland.) Available online at: http://www.redlist.org [verified 20 February 2006].

Kaemmerer K. (2003). Ocelot (Leopardus pardalis) Species Survival Plan Report. Dallas Zoo, Dallas, TX. Available online at: www.aza.org

Makler, A. , David, R. , Blumenfield, Z. , and Better, O. S. (1981). Factors affecting sperm motility VII. Sperm viability as affected by change of pH and osmolarity of semen and urine specimens. Fertil. Steril. 36, 507–511.
| PubMed | Ministerio do Meio Ambiente (MMA) (2003). ‘National List of Endangered Brazilian Wildlife Species.’ (Instuto Brasileiro do Meio Ambiente e dos Recursos Naturais Renovaveis: Brasilia.) Available online at: http://www.mma.gov.br [verified 20 February 2006].

Moraes W., Morais R. N., Moreira N., Lacerda O., Gomes M. L. F., Mucciolo R. G., and Swanson W. F. (1997). Successful artificial insemination after exogenous gonadotropin treatment in the ocelot (Leopardus pardalis) and tigrina (Leopardus tigrina). In ‘Proceedings of the American Association of Zoo Veterinarians Annual Meeting, 1997, Houston, TX’. pp. 334–335.

Morais, R. N. , Mucciolo, R. G. , Gomes, M. L. F. , Lacerda, O. , Moraes, W. , Moreira, N. , Graham, L. H. , Swanson, W. F. , and Brown, J. L. (2002). Seasonal analysis of seminal characteristics, serum testosterone and fecal androgen in the ocelot (Leopardus pardalis), margay (L. wiedii) and tigrina (L. tigrinus). Theriogenology 57, 2027–2041.
| Crossref | GoogleScholarGoogle Scholar | PubMed | Munro C. J., and Lasley B. L. (1988). Non-radiometric methods for immunoassay of steroid hormones. In ‘Non-radiometric Assays: Technology and Application in Polypeptide and Steroid Hormone Detection’. (Eds B. D. Albertson and F. P. Haseltine.) pp. 289–329. (Alan R. Liss: New York.)

Platz, C. C. , Wildt, D. E. , and Seager, S. W. J. (1978). Pregnancy in the domestic cat after artificial insemination with previously frozen spermatozoa. J. Reprod. Fert. 52, 279–282.
Sunquist M., and Sunquist F. (2002). ‘Wilds Cats of the World.’ (The University of Chicago Press: Chicago, IL.)

Swanson W. F. (2001). Reproductive biotechnology and conservation for the forgotten felids-the small cats. In ‘Proceedings of the 1st International Symposium in Assisted Reproductive Technology for Conservation and Genetic Management of Wildlife, Omaha, NE’. pp. 100–120.

Swanson, W. F. (2003a). Research in nondomestic species: experiences in reproductive physiology research for conservation of endangered felids. ILAR J. 44(4), 307–316.
| PubMed | Swanson W. F. (2003b). The role of science and reproductive biotechnology in establishing and managing the Brazilian ocelot population in US and Brazilian zoos. In ‘Proceedings of the American Zoo and Aquarium Association’s 2003 Annual Conference, 7–11 September 2002, Cincinnati, OH’. (Ed. C. K. Baer.) pp. 75–78.

Swanson, W. F. (2006). Application of assisted reproduction for population management in felids: the potential and reality for conservation of small cats. Theriogenology 66, 49–58.
| Crossref | GoogleScholarGoogle Scholar | PubMed | Tewes M. E., and Schmidly D. J. (1987). The neotropical felids: jaguar, ocelot, margay, and jagurundi. In ‘Wild Furbearer Management and Conservation in North America’. (Eds M. Novak, J. A. Baker, M. E. Obbard and B. Malloch.) pp. 697–711. (Ontario Ministry of Natural Resources: Toronto.)

Tewes M. E., and Everett D. D. (1986). Status and distribution of the endangered ocelot and jagurundi in Texas. In ‘Cats of the World: Biology, Conservation and Management’. (Eds S. D. Miller and D. D. Everett.) pp. 147–158. (National Wildlife Federation: Washington, DC.)

Thiangtum, K. , Swanson, W. F. , Howard, J. , Tunwattana, W. , Tongthainan, D. , Wichasilpa, W. , Patumrattanathan, P. , and Pinyopoommintr, T (2006). Assessment of basic seminal characteristics, sperm cryopreservation and heterologous in vitro fertilization in the fishing cat (Prionailurus viverrinus).  Reprod. Fertil. Develop. 18, 373–382.
| Crossref | GoogleScholarGoogle Scholar | Wildt D. E. (1994). Endangered species spermatozoa: diversity, research and conservation. In ‘Function of Somatic Cells in the Testes’. (Ed. A. Bartke.) pp. 1–24. (Springer-Verlag: New York.)

Wildt, D. E. , O’Brien, S. J. , Howard, J. G. , Caro, T. M. , Roelke, M. E. , Brown, J. L. , and Bush, M. (1987). Similarity in ejaculate-endocrine characteristics in captive versus free-living cheetahs of two subspecies. Biol. Reprod. 36, 351–360.
| Crossref | GoogleScholarGoogle Scholar | PubMed | USDI Fish and Wildlife Service (1982). Endangered and Threatened Wildlife and Plants; Endangered Status for U.S. Population of the Ocelot, Final Rule. Federal Register 47(140): 31670–31672.