Giraffes are numbered amongst vulnerable mammalian species which are likely to become endangered due to the ongoing population decline. In endangered species, the ex situ biodiversity conservation efforts involve captive breeding, genome resource banking and artificial reproduction using cryopreserved gametes. Determination of the sperm standards and the sensitivity of spermatozoa to freezing are crucial points in such activities. We analysed epididymal sperm retrieved from three captive-bred giraffe males (two Giraffa reticulata, one Giraffa camelopardalis rothschildi) and tested the sensitivity of giraffe sperm to cryopreservation. The basic testicular and sperm morphometric measurements, the sperm functional and morphological assessment and the sperm chromatin structure analysis revealed interindividual differences. The epididymal sperm quality of males G2 and G3 was low, probably due to the long-term health problems in G2, sexual immaturity in G3 and longer period between the death/neutering and the sperm retrieval in both animals. Epididymal sperm of male G1 was subjected to cryopreservation in TRIS-fructose-citric acid media with egg yolk. The tested cryopreservation method was approved as suitable for banking giraffe spermatozoa. In the case of endangered species, euthanized and neutered animals can serve as a source of gametes for genome resource banking.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Price excludes VAT (USA)
Tax calculation will be finalised during checkout.
Abaigar T, Cano M, Pickard AR, Holt WV (2001) Use of computer-assisted sperm motility assessment and multivariate pattern analysis to characterize ejaculate quality in Mohor gazelles (Gazella dama mhorr): effects of body weight, electroejaculation technique and short-term semen storage. Reprod 122:265–273
Agarwal A, Said TM (2003) Role of sperm chromatin abnormalities and DNA damage in male infertility. Hum Reprod Update 9:331–345
Ballachey BE, Hohenboken WD, Evenson DP (1987) Heterogeneity of sperm nuclear chromatin structure and its relationship to bull fertility. Biol Reprod 36:915–925
Ballachey BE, Evenson DP, Saacke RG (1988) The sperm chromatin structure assay. Relationship with alternate tests of semen quality and heterospermic performance of bulls. J Androl 9:109–115
Bock F, Fennessy J, Bidon T, Tutchings A, Marais A, Deacon F, Janke A (2014) Mitochondrial sequences reveal a clear separation between Angolan and South African giraffe along a cryptic rift valley. BMC Evol Biol 14:219. https://doi.org/10.1186/s12862-014-0219-7
Boe-Hansen GB, Christensen P, Vibjerg D, Nielsen MBF, Hedeboe AM (2008) Sperm chromatin structure integrity in liquid stored boar semen and its relationships with field fertility. Theriogenology 69:728–736. https://doi.org/10.1016/j.theriogenology.2007.12.004
Casaretto C, Lombardo DM, Giuliano S, et al (2012) Morphometric analysis of llama (Lama glama) sperm head. Andrologia 44:424–430
Cassinello J, Abaigar T, Gomendio M, Roldan ER (1998) Characteristics of the semen of three endangered species of gazelles (Gazella dama mhorr, G. Dorcas neglecta and G. Cuvieri). J Reprod Fertil 113:35–45
Chan PJ, Corselli JU, Jacobson JD, Patton WC, King A (1996) Correlation between intact sperm acrosome assessed using the Spermac stain and sperm fertilizing capacity. Arch Androl 36:25–27
Comizzoli P (2015) Biobanking efforts and new advances in male fertility preservation for rare and endangered species. Asian J Androl 17:640–645. https://doi.org/10.4103/1008-682X.153849
Comizzoli P, Wildt DE (2013) Mammalian fertility preservation through cryobiology: value of classical comparative studies and the need for new preservation options. Reprod Fertil Dev 26:91–98. https://doi.org/10.1071/RD13259
Comizzoli P, Songsasen N, Hagedorn M, Wildt DE (2012) Comparative cryobiological traits and requirements for gametes and gonadal tissues collected from wildlife species. Theriogenology 78:1666–1681. https://doi.org/10.1016/j.theriogenology.2012.04.008
Cummins JM, Woodall PF (1985) On mammalian sperm dimensions. J Reprod Fertil 75:153–175. https://doi.org/10.1530/jrf.0.0750153
Erenpreiss J, Spano M, Erenpreisa J, Bungum M, Giwercman A (2006) Sperm chromatin structure and male fertility: biological and clinical aspects. Asian J Androl 8:11–29. https://doi.org/10.1111/j.1745-7262.2006.00112.x
Evenson DP (1999) Loss of livestock breeding efficiency due to uncompensable sperm nuclear defects. Reprod Fertil Dev 11:1–15
Evenson DP, Wixon R (2006) Clinical aspects of sperm DNA fragmentation detection and male infertility. Theriogenology 65:979–991. https://doi.org/10.1016/j.theriogenology.2005.09.011
Evenson DP, Jost LK, Marshall D et al (1999) Utility of the sperm chromatin structure assay as a diagnostic and prognostic tool in the human fertility clinic. Hum Reprod 14:1039–1049
Fennessy J, Bidon T, Reuss F, Kumar V, Elkan P, Nilsson MA, Vamberger M, Fritz U, Janke A (2016) Multi-locus analyses reveal four giraffe species instead of one. Curr Biol 26:2543–2549. https://doi.org/10.1016/j.cub.2016.07.036
Gage MJG (1998) Mammalian sperm morphometry. Proc R Soc Lond B Biol Sci 265:97–103. https://doi.org/10.1098/rspb.1998.0269
Gage MJG, Freckleton RP (2003) Relative testis size and sperm morphometry across mammals: no evidence for an association between sperm competition and sperm length. Proc R Soc Lond B Biol Sci 270:625–632. https://doi.org/10.1098/rspb.2002.2258
Garde JJ, Soler AJ, Cassinello J, Crespo C, Malo AF, Espeso G, Gomendio M, Roldan ERS (2003) Sperm cryopreservation in three species of endangered gazelles (Gazella cuvieri, G. Dama mhorr, and G. Dorcas neglecta). Biol Reprod 69:602–611. https://doi.org/10.1095/biolreprod.102.012914
Gillan L, Kroetsch T, Maxwell WMC, Evans G (2008) Assessment of in vitro sperm characteristics in relation to fertility in dairy bulls. Anim Reprod Sci 103:201–214. https://doi.org/10.1016/j.anireprosci.2006.12.010
Gilmore JA, McGann LE, Ashworth E et al (1998) Fundamental cryobiology of selected African mammalian spermatozoa and its role in biodiversity preservation through the development of genome resource banking. Anim Reprod Sci 53:277–297
Hall-Martin AJ, Skinner JD, Hopkins BJ (1978) The development of the reproductive organs of the male giraffe, Giraffa camelopardalis. J Reprod Fertil 52:1–7
Kaneko T, Ito H, Sakamoto H, Onuma M, Inoue-Murayama M (2014) Sperm preservation by freeze-drying for the conservation of wild animals. PLoS One 9:e113381. https://doi.org/10.1371/journal.pone.0113381
Larson KL, DeJonge CJ, Barnes AM, Jost LK, Evenson DP (2000) Sperm chromatin structure assay parameters as predictors of failed pregnancy following assisted reproductive techniques. Hum Reprod 15:1717–1722
Leibo SP, Songsasen N (2002) Cryopreservation of gametes and embryos of non-domestic species. Theriogenology 57:303–326
Lueders I, Niemuller C, Pootoolal J, Rich P, Gray C, Streich WJ, Hildebrandt TB (2009) Sonomorphology of the reproductive tract in male and pregnant and non-pregnant female Rothschild’s giraffes (Giraffa camelopardalis rotschildi). Theriogenology 72:22–31. https://doi.org/10.1016/j.theriogenology.2009.01.016
Lüpold S, Manier MK, Puniamoorthy N, Schoff C, Starmer WT, Luepold SHB, Belote JM, Pitnick S (2016) How sexual selection can drive the evolution of costly sperm ornamentation. Nature 533:535–538. https://doi.org/10.1038/nature18005
Pablos MTP, Saragusty J, Santiago-Moreno J, Stagegaard J, Göritz F, Hildebrandt TB, Hermes R (2015) Cryopreservation of Onager (Equus hemiounus onager) epididymal spermatozoa. J Zoo Wildl Med 46:517–525. https://doi.org/10.1638/2014-0243.1
Roldan ERS, Gomendio M, Vitullo AD (1992) The evolution of eutherian spermatozoa and underlying selective forces: female selection and sperm competition. Biol Rev 67:551–593. https://doi.org/10.1111/j.1469-185X.1992.tb01193.x
Roldan ER, Cassinello J, Abaigar T, Gomendio M (1998) Inbreeding, fluctuating asymmetry, and ejaculate quality in an endangered ungulate. Proc Biol Sci 265:243–248. https://doi.org/10.1098/rspb.1998.0288
Roldan ERS, Gomendio M, Garde JJ, Espeso G, Ledda S, Berlinguer F, del Olmo A, Soler AJ, Arregui L, Crespo C, González R (2006) Inbreeding and reproduction in endangered ungulates: preservation of genetic variation through the Organization of Genetic Resource Banks. Reprod Domest Anim 41(Suppl 2):82–92. https://doi.org/10.1111/j.1439-0531.2006.00772.x
Roth TL, Stoops MA, Robeck TR, O’Brien JK (2016) Factors impacting the success of post-mortem sperm rescue in the rhinoceros. Anim Reprod Sci 167:22–30. https://doi.org/10.1016/j.anireprosci.2016.01.019
Rubio C, Gil-Salom M, Simón C et al (2001) Incidence of sperm chromosomal abnormalities in a risk population: relationship with sperm quality and ICSI outcome. Hum Reprod 16:2084–2092
Ruiz-Lopez MJ, Evenson DP, Espeso G, Gomendio M, Roldan ERS (2010) High levels of DNA fragmentation in spermatozoa are associated with inbreeding and poor sperm quality in endangered ungulates. Biol Reprod 83:332–338. https://doi.org/10.1095/biolreprod.110.084798
Ruiz-López MJ, Gañan N, Godoy JA et al (2012) Heterozygosity-fitness correlations and inbreeding depression in two critically endangered mammals. Conserv Biol 26:1121–1129. https://doi.org/10.1111/j.1523-1739.2012.01916.x
Rybar R, Markova P, Veznik Z, Faldikova L, Kunetkova M, Zajicova A, Kopecka V, Rubes J (2009) Sperm chromatin integrity in young men with no experiences of infertility and men from idiopathic infertility couples. Andrologia 41:141–149. https://doi.org/10.1111/j.1439-0272.2008.00905.x
Saragusty J, Gacitua H, King R, Arav A (2006) Post-mortem semen cryopreservation and characterization in two different endangered gazelle species (Gazella gazella and Gazella dorcas) and one subspecies (Gazella gazelle acaiae). Theriogenology 66:775–784. https://doi.org/10.1016/j.theriogenology.2006.01.055
Strand J, Ragborg MM, Pedersen HS, Kristensen TN, Pertoldi C, Callesen H (2016) Effects of post-mortem storage conditions of bovine epididymides on sperm characteristics: investigating a tool for preservation of sperm from endangered species. Conserv Physiol 4:cow069. https://doi.org/10.1093/conphys/cow069
Thurston LM, Watson PF (2002) Semen cryopreservation: a genetic explanation for species and individual variation? Cryoletters 23:255–262
Velhankar DP, Hukeri VB, Deshpande BR, Sane CR (1973) Biometry of the genitalia and the spermatozoa of a male giraffe. Indian Vet J 50:789–792
Waterhouse KE, Haugan T, Kommisrud E, Tverdal A, Flatberg G, Farstad W, Evenson DP, de Angelis PM (2006) Sperm DNA damage is related to field fertility of semen from young Norwegian red bulls. Reprod Fertil Dev 18:781–788
Yániz JL, Soler C, Santolaria P (2015) Computer assisted sperm morphometry in mammals: a review. Anim Reprod Sci 156:1–12. https://doi.org/10.1016/j.anireprosci.2015.03.002
The authors are grateful to J. Vahala and L. Pavlacik from Dvur Kralove nad Labem Zoo, to J. Vodicka from Prague Zoo and the staff of Bratislava Zoo for providing the samples. We acknowledge Mr. Paul Veater (Bristol, United Kingdom) for proofreading the translated manuscript.
This work was supported by the Ministry of Education, Youth and Sports of the Czech Republic under the project CEITEC 2020 (LQ1601) and by the Ministry of Agriculture of the Czech Republic (RO 0518).
The study complies with the current laws of the Czech Republic. All institutional and national guidelines for the care and use of animals were followed.
Conflict of interests
The authors declare that they have no conflict of interest.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Sipek, J., Vozdova, M., Prinosilova, P. et al. Sperm and testicular measurements and sperm cryopreservation in the giraffe (Giraffa). Eur J Wildl Res 65, 16 (2019). https://doi.org/10.1007/s10344-019-1257-7
- Chromatin integrity